Antinociceptive effect of leaf essential oil from Croton sonderianus in mice

The leaf essential oil from Croton sonderianus (EOCS) was evaluated for antinociceptive activity in mice using chemical and thermal models of nociception. Given orally, the essential oil at doses of 50, 100 and 200 mg/kg produced significant inhibitions on chemical nociception induced by intraperitoneal acetic acid and subplantar formalin or capsaicin injections. However, it evidenced no efficacy against thermal nociception in hot-plate test. More prominent inhibition of acetic acid-induced writhing and capsaicin-induced hind-paw licking responses was observed at 100 and 200 mg/kg of EOCS. At similar doses, the paw licking behavior in formalin test was more potently suppressed during the late phase (20-25 min, inflammatory) than in early phase (0-5 min, neurogenic). The EOCS-induced antinociception in both capsaicin and formalin tests was insensitive to naloxone (1 mg/kg, s.c.), but was significantly antagonized by glibenclamide (2 mg/kg, i.p.). In mice, the essential oil (100 and 200 mg/kg) neither significantly enhanced the pentobarbital-sleeping time nor impaired the motor performance in rota-rod test, indicating that the observed antinociception is unlikely due to sedation or motor abnormality. These results suggest that EOCS produces antinociception possibly involving glibenclamide-sensitive KATP+ channels, which merit further studies on its efficacy in more specific models of hyperalgesia and neuropathic pain.     

Genetic differentiation among three neighboring Brazil-cherry (Eugenia uniflora L.) populations within the Brazilian Atlantic rain forest

Eugenia uniflora L. (pitanga) is widely distributed in tropical areas. It is present in coastal vegetation from Ceara, in northeastern Brazil, to Rio Grande do Sul, at the southern tip of the country. Eugenia uniflora is of ecological importance, both as colonizing species on disturbed land and as food supplier for a wide variety of insects, birds and mammals. Pitanga plays a role in the maintenance of shrubby coastal ecosystems, especially at disturbed sites, and in 'restinga' ecosystems, at the interface between low forest and strand vegetation. To investigate the genetic diversity residing within the species, three neighboring populations at a distance of less than 24 km from each other, with varying degrees of human impact, were studied. The level of genetic diversity within and between populations was assessed with amplified fragment length polymorphism (AFLP) methodology. A total of 532 AFLP markers were analyzed in 66 individual trees. The polymorphism level varied from 61.2 to 96% depending on the primer combination used. Intra- and inter-population genetic diversity analysis showed that more than 88% of the variation resided within the populations, with a G_st of 0.123. Nevertheless, using neighbor joining (NJ) and principal component analysis (PCA), on the genetic distance (GD) data, permitted the three analyzed populations to be differentiated.     

Anti-Trypanosoma cruzi and cytotoxic activities of Eugenia uniflora L

Chagas disease is caused by Trypanosoma cruzi, being considered a public health problem. An alternative to combat this pathogen is the use of natural products isolated from fruits such as Eugenia uniflora, a plant used by traditional communities as food and medicine due to its antimicrobial and biological activities. Ethanolic extract from E. uniflora was used to evaluate in vitro anti-epimastigote and cytotoxic activity. This is the first record of anti-Trypanosoma activity of E. uniflora, demonstrating that a concentration presenting 50% of activity (EC(50)) was 62.76 μg/mL. Minimum inhibitory concentration (MIC) was ≤ 1024 μg/mL. Our results indicate that E. uniflora could be a source of plant-derived natural products with anti-epimastigote activity with low toxicity.     

Purification of a lectin from Eugenia uniflora L. seeds and its potential antibacterial activity

Aims: The aim of this work was to analyse the antimicrobial properties of a purified lectin from Eugenia uniflora L. seeds. Methods and Results: The E. uniflora lectin (EuniSL) was isolated from the seed extract and purified by ion‐exchange chromatography in DEAE‐Sephadex with a purification factor of 11·68. The purified lectin showed a single band on denaturing electrophoresis, with a molecular mass of 67 kDa. EuniSL agglutinated rabbit and human erythrocytes with a higher specificity for rabbit erythrocytes. The haemagglutination was not inhibited by the tested carbohydrates but glycoproteins exerted a strong inhibitory action. The lectin proved to be thermo resistant with the highest stability at pH 6·5 and divalent ions did not affect its activity. EuniSL demonstrated a remarkable nonselective antibacterial activity. EuniSL strongly inhibited the growth of Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella sp. with a minimum inhibitory concentration (MIC) of 1·5 μg ml^?1, and moderately inhibited the growth of Bacillus subtilis, Streptococcus sp. and Escherichia coli with a MIC of 16·5 μg ml^?1. Conclusions: EuniSL was found to be effective against bacteria. Significance and Impact of the Study: The strong antibacterial activity of the studied lectin indicates a high potential for clinical microbiology and therapeutic applications.     

Synthesis and Characterization of Eugenia uniflora L. Silver Nanoparticles and L-Cysteine Sensor Application

L-Cysteine (Cys) is a non-essential sulfur-containing amino acid, crucial for protein synthesis, detoxification, and several metabolic functions. Cys is widely used in the agricultural, food, cosmetic, and pharmaceutical industries. So, a suitable sensitive and selective sensing approach is of great interest, and a low-cost sensor would be necessary. This article presents silver nanoparticles (EuAgNPs) synthesized by a green synthesis method using Eugenia uniflora L. extracts and photoreduction. The nanoparticles were characterized by UV/VIS, transmission electron microscopy, high-performance liquid chromatography (HPLC), FTIR, and Zeta potential. With the addition of Cys in the EuAgNPs solution, the terminal thiol part of L-cysteine binds on the surface of nanoparticles through Ag−S bond. The EuAgNPs and CysAgNPs coexist until flavonoids bound the amino group of Cys, enhancing the red color of solutions. The EuAgNPs provided selectivity to detect Cys among other amino acids, and its detection limit was found to be 3.8 nM. The sensor has the advantages of low-cost synthesis, fast response, high selectivity, and sensitivity.     

The taxonomic significance of leaf micromorphology in the genus Eugenia L. (Myrtaceae)

Leaf micromorphological features of both the Old and New World representatives of Eugenia L. have been studied by light microscopy and scanning electron microscopy. The leaves are hypostomatic and, in the majority of species, the stomata are raised above the epidermis. Trichomes are usually present on the abaxial surface of the New World species. The patterns of abaxial surface sculpturing and arrangement of outer stomatal rims are unique in the New World species. The majority of the Old W'orld species showed different anticlinal wall patterns on the adaxial and abaxial epidermises. In the majority of the New World species the anticlinal wall patterns are more or less the same except in E. axillaris. The paracytic type of stomata, with a non-prominent T-piece cutinization at the poles of the guard cells, is a characteristic feature of the New World species. The taxonomic significance of these features in identification and elucidation of species from both areas is discussed.     

Impact of iron toxicity on oxidative metabolism in young Eugenia uniflora L. plants

In excess, iron can induce the production and accumulation of reactive oxygen species (ROS), causing oxidative stress. The objective of this work was to evaluate the impact of toxic concentrations of iron (Fe) on the antioxidative metabolism of young Eugenia uniflora plants. Forty-five-day-old plants grown in Hoagland nutrient solution, pH 5.0, were treated with three Fe concentrations, in the form of FeEDTA, during three periods of time. At the end of the treatment, the plants were harvested and relative growth rate, iron content, lipid peroxidation and enzymes and metabolites of the antioxidative metabolism were determined. Iron-treated plants showed higher iron contents, reduced relative growth rates and iron toxicity symptoms in both leaves and roots. There was an increase in lipid peroxidation with increasing Fe, only in the leaves. The enzymatic activities of superoxide dismutase (SOD) and glutathione reductase (GR) increased with increasing Fe concentration and treatment exposure time. The activities of catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APX) also increased with increasing Fe concentration but decreased with increasing treatment exposure time. Glutathione peroxidase activity (GPX) decreased with increasing Fe concentration and exposure time. The ascorbate (AA) and reduced glutathione (GSH) contents and the AA/DHA and GSH/GSSG ratios, in general, increased with increasing Fe concentration and treatment exposure time. The results indicate that under toxic levels of Fe, young E. uniflora plants suffer increased oxidative stress, which is ameliorated through changes in the activities of antioxidative enzymes and in the contents of the antioxidants AA and GSH.     

Antiproliferative and cytotoxic effects of purple pitanga (Eugenia uniflora L.) extract on activated hepatic stellate cells

The presence of phenolic compounds in fruit‐ and vegetable‐rich diets has attracted researchers' attention due to their health‐promoting effects. The objective of this study was to evaluate the effects of purple pitanga (Eugenia uniflora L.) extract on cell proliferation, viability, mitochondrial membrane potential, cell death and cell cycle in murine activated hepatic stellate cells (GRX). Cell viability by 3‐(4,5‐dimethylthiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) assay was significantly decreased on cells treated with 50 and 100 µg ml^?1 of purple pitanga extract for 48 and 72 h, and the percentage of dead cell stained with 7‐amino‐actinomycin D was significantly higher in treated cells. The reduction of cell proliferation was dose dependent, and we also observed alterations on cell cycle progression. At all times studied, GRX cells treated with 50 and 100 µg ml^?1 of purple pitanga showed a significant reduction in cellular mitochondrial content as well as a decrease in mitochondrial membrane potential. Furthermore, our results indicated that purple pitanga extract induces early and late apoptosis/necrosis and necrotic death in GRX cells. This is the first report describing the antiproliferative, cytotoxic and apoptotic activity for E. uniflora fruits in hepatic stellate cells. The present study provides a foundation for the prevention and treatment of liver fibrosis, and more studies will be carried to elucidate this effect. Copyright © 2013 John Wiley & Sons, Ltd.     

Antinociceptive effects of the essential oil of Alpinia zerumbet on mice.

Alpinia zerumbet (Pers.) Burtt. et Smith is an aromatic plant that is distributed widely in the tropical and sub-tropical regions of the world. In Brazil, where A. zerumbet is called "colonia", it is used widely in folk medicine for the treatment of various diseases, including hypertension. In the present study, the antinociceptive effects of the orally administered essential oil of A. zerumbet (EOAz) were evaluated in male Swiss mice (20-25 g each). In the acetic acid-induced writhing test, EOAz (30, 100 and 300 mg/kg body wt.; n = 10, n = 13 and n = 15, respectively) was effective at all doses. In the hot-plate test, EOAz significantly increased the latency at doses of 100 and 300 mg/kg body wt., but not at 30 mg/kg body wt., at all observation times up to the 180th min (n = 10 for each dose). In the formalin test, EOAz significantly reduced paw licking time in the second phase of the test at 100 mg/kg body wt. (n = 10), but decreased it in both phases at 300 mg/kg body wt. (n = 10). At 30 mg/kg body wt., the effect of EOAz did not differ from control values in either phase of the formalin test (n = 10). Pretreatment with naloxone (5 mg/kgbodywt., i.p.) caused a significant reversal of the analgesic effect of 300 mg/kg body wt. EOAz (n = 8) that was complete for the first phase, but only partial for the second phase of the formalin test. The data show that orally administered OEAz promotes a dose-dependent antinociceptive effect, with a mechanism of action which probably involves the participation of opiate receptors.     

Leaf anatomical studies of Eugenia L. (Myrtaceae) species from the Malay Peninsula

KHATIJAH, H. H., CUTLER, D. F. & MOORE, D. M., 1992. Leaf anatomical studies of Eugenia L. species from the Malay Peninsula. The leaf and petiole anatomy of 25 Malaysian species of Eugenia L. sensu lato was investigated to determine the value of vegetative anatomy for the identification of non-flowering material. Twelve characters proved useful for the identification of species: type of stomata; shape of leaves in T.S.; shape of midrib bundle; sclerenchyma sheath; cutinization of outer epidermal wall; presence or absence of sclereids, idioblasts, hypodermis, columnar epidermal cells, solitary crystals; number of palisade layers and the shape of vascular strand, sclerenchyma sheath and sclereids in the parenchyma of the petiole. Using these characters an anatomical key was constructed to assist identification of the species. Only the closely related species E. stapfiana and E. nilidula could not be separated on anatomical characters.
The anatomical data support the treatment of Henderson (1949) of section Acmena but not of sections Cleistocalyx and Fissicalyx , which fit better in section Syzygium.     

Bioactivity of two major constituents isolated from the essential oil of Artemisia judaica L

The essential oil of Artemisia judaica L., grown on Sinai Peninsula of Egypt, was extracted via hydrodistillation. Chromatographic separation on repeated silica gel columns led to isolate two compounds namely piperitone and trans-ethyl cinnamate. Insecticidal, antifeedant and antifungal properties of the isolated compounds were examined. Both compounds showed pronounced insecticidal and antifeedant activity against the third instar larvae of Spodoptera littoralis (Boisd). trans-Ethyl cinnamate (LD₅₀ = 0.37 μg/larva) was more toxic than piperitone (LD₅₀ = 0.68 μg/larva). The two isolated compounds revealed antifeedant activity in a concentration dependent manner, with complete feeding inhibition at a concentration of 1000 μg/ml. When tested for antifungal activity against four plant pathogenic fungi, the isolated compounds exhibited a moderate to high activity.     

Studies on the acute and sub-chronic toxicity of the essential oil of Ocimum gratissimum L. leaf.

Oral and intra-peritoneal acute toxicity and the sub-chronic intra-peritoneal toxicity of the essential oil of Ocimum gratissimum Linn, Lamiaceae (Ocimum oil), was investigated. The acute toxicity test involved the oral and intra-peritoneal administration of graded doses of Ocimum oil prepared as a 4% v/v emulsion to 2 groups each of 30 rats and mice. LD50 and LD100 were determined for both routes and species. In the sub-chronic toxicity study, 25 male Sprague-Dawley rats were randomized into 4 test groups (treated with three graded sub-lethal doses of Ocimum oil prepared as a 4% v/v emulsion) and a control. Organs and blood samples were taken for analyses after a 30 day treatment period. A dose-dependent sedative effect of Ocimum oil was observed during the acute toxicity study in mice and rats and in the sub-chronic test in rats. Evidence of treatment, route, and dose-dependent toxicity were detected in both studies. Changes in weight of the testes, hearts, kidneys, intestines and lungs of the rats were statistically insignificant (ANOVA P < 0.05). Data analyses of blood biochemical, haematological and histopathological findings showed significant differences between control and treated groups and revealed that Ocimum oil is capable of invoking an inflammatory response that transits from acute to chronic on persistent administration. While the study revealed that Ocimum oil might be better tolerated when administered orally for systemic delivery, the oil has toxic potentialities that should not be overlooked.     

Geographical ecology and conservation of Eugenia L. (Myrtaceae) in the Brazilian Cerrado: Past,present and future

Human actions have caused the fragmentation of natural vegetation, habitat loss and climate change. The Cerrado, considered one of the global hotspots of diversity, has suffered great habitat loss due to these factors, which has been aggravated by the agricultural expansion that took place during the last 60 years. In this context, we chose species of the genus Eugenia L. (Myrtaceae) occurring in the Brazilian Cerrado to describe richness patterns and range loss, and determine conservation priorities for the Cerrado. Ecological niche models (ENMs) were applied to calculate the geographical range of each species in the past (Last Glacial Maximum – LGM, 21 000 years ago), present (PIP, representing current climatic conditions – 1760 years ago) and future (near future – NF, 2080–2100). These results were combined to calculate the richness of the group and also to estimate the range loss of these species in the future. Moreover, we evaluated the irreplaceability of areas for species conservation, aiming to maximize the biotic stability of Eugenia species. Our results showed that the highest species richness in the past was found in the southwestern region of the Cerrado and, currently, the richest regions are found in the central and southeastern areas. However, in the future, we predict a shift of the greatest values of richness towards the southeastern region, an area currently occupied by the Atlantic forest. Although areas with high conservation priorities were found scattered across the biome, this shift is worrisome due to the high fragmentation rate and intensive human occupation thoughout the Atlantic region. Thus, conservation efforts should focus on areas found within these limits.     

Even population differentiation for maternal and biparental gene markers in Eugenia uniflora, a widely distributed species from the Brazilian coastal Atlantic rain forest

Brazilian cherry (Eugenia uniflora L.) occurs in the Brazilian Atlantic Coastal Tropical Forest from the north‐eastern state of Ceara, to the southernmost state of Rio Grande do Sul. E. uniflora plays an important role in the maintenance of ‘Restinga’ ecosystems, at the interface between forest and strand vegetation. Here, we characterize the genetic diversity, its allocation within and between populations and the possible components of nuclear and cytoplasmic gene flow that determine the spatial distribution of the genetic variability. Five E. uniflora populations encompassing distinct biogeographical components were sampled: populations at extremes of latitudinal distribution, at an oceanic island and close to the major urban centre of Rio de Janeiro. AFLP markers showed that genetic variance among studied populations was moderate (F_st = 0.211) with 78.9% of variability residing within populations. The estimated seed and pollen components of gene flow among populations were approximately equal, with a ratio between 1.003 and 0.713 in function of the cpDNA marker used. The five populations present a considerable genetic structure, as assigned by both nuclear and chloroplastic DNA markers. Our data suggest the existence of different glacial refugia and a limited pollen and seed gene flow, mainly between the southern and the other regions enclosed in the Atlantic rain forest. Therefore, any strategy of conservation and management has to assure the preservation of several populations along the Atlantic coast, to maintain the majority of the intra specific level of diversity.     

Chemical composition and anticancer activity of leaf essential oil of Myrica gale L.

Myrica gale L. (Myricaceae), a native plant from Canada used in traditional medicine, was extracted by hydrodistillation and the oil was collected after 30 and 60 min. The chemical composition of these two extracts was determined using GC-MS analysis. We identified 53 components and myrcene (23.18-12.14%), limonene (11.20-6.75%), alpha-phellandrene (9.90-6.49%) and beta-caryophyllene (9.31-10.97%) were the major components in the 30- and 60-min fractions, respectively, whereas higher caryophyllene oxide content was detected in the 60-min fraction (9.94%) than in the 30-min fraction (3.47%). The anticancer activities of these extracts were assessed against human lung carcinoma cell line A-549 and human colon adenocarcinoma cell line, DLD-1. The 60-min fraction showed higher anticancer activity against both tumor cell lines with an IC50 value of 88 +/- 1 microg/ml. The 30-min fraction had an IC50 value of 184 +/- 4 microg/ml for A-549 and 160 +/- 3 microg/ml for DLD-1. The higher cell growth inhibition induced by the 60-min fraction, as compared to the 30-min fraction, could be due to sesquiterpene enrichment.     

Variation in the essential oils composition in Brazilian populations of Schinus molle L. (Anacardiaceae)

Schinus molle L. is popularly known as “peppertree” and presents high essential oil contents with a pleasant spicy scent. Its fruits are used in beverages or dried and ground as a substitute for pepper. Air-dried samples of 11 Brazilian populations of S. molle were extracted using steam distillation and analyzed using GC and GC–MS. A total of 22 volatile compounds were detected and identified. Four groups were formed by the Average Linkage cluster analysis. The first group was characterized by the compound sabinene; the second, which was the largest group, was characterized by the presence of α- and β-pinene; the third group comprised the samples corresponding to the São Borja population and was characterized by the high contents of α-cadinol; and the fourth group was characterized by the high concentrations of myrcene.     

Phylogenetic relationships of the genus Chamaecyparis inferred from leaf essential oil

The species differentiation between Chamaecyparis formosensis, C. obtusa var. formosana, and C. obtusa, based on the composition of the leaf essential oils, was studied. The characterization of the oils by GC-FID and GC/MS analyses showed remarkable differences between these three essential oils. Cluster analysis (CA) and principal-component analysis (PCA) distinguished three groups of essential oils. The C. formosensis oil was dominated by α-pinene while those isolated from C. obtusa var. formosana and C. obtusa were characterized by high levels of (-)-thujopsene and α-terpinyl acetate, respectively. Moreover, the phylogenetic relationships of the genus Chamaecyparis were in agreement with previous findings based on morphological and molecular evidence. In addition, the essential oils from C. obtusa var. formosana could be classified into three chemical types, according to their different characteristic main compounds (β-elemol, (-)-thujopsene, and cis-thujopsenal). The biochemical correlations between the major constituents of the Chamaecyparis species were examined and their relationship is discussed.     

Foliar anatomy and micromorphology of eleven species of Eugenia L. (Myrtaceae)

The anatomy and micromorphology of eleven species of Eugenia found in the 'restinga' of Marica, Rio de Janeiro, Brazil, were studied and compared. The characteristics found to be of most use in distinguishing the species are: presence and types of trichomes persistent in the adult material, form and arrangement of overlying cells (epidermal cells over secretory cavities), pattern of anticlinal walls of epidermal cells (primary sculpturing), occurrence of epicuticular striae and folds (secondary sculpturing) and presence of peristomatal folds. An analytical key based on these characteristics is presented for the Eugenia species studied.     

辽东(木匆心)木挥发油的成分

本文采用气相色谱-质谱-计算机（GC-MS-DS）联用技术,首次研究了辽东楤木根皮中的挥发性成分,鉴定了37个化合物。成分中以α-姜黄烯（α-curcumene）等为主,含量达15.32％,此外尚含有薁类和大量的萘衍生物。