Anti-hyperglycemic fraction from Alternanthera sessilis L. leaves gets elucidated following bioassay-guided isolation and mass spectrometry

Abstract The anecdotal use of Alternanthera sessilis L. as a relief for diabetes has been known in the Philippines for generations, and antidiabetic activity of similar varieties in other countries is likewise documented. However, the compounds responsible for this activity remain unclear. This study aims to isolate the anti-hyperglycemic fraction of local A. sessilis leaves and identify the compounds in this fraction. Methanol extract of A. sessilis leaves and its hexane, ethyl acetate (ASE), and water fractions were administered to alloxan-induced diabetic mice. ASE (250mg/kg) had the highest anti-hyperglycemic activity at 6-h post-treatment (25.81%±12.72%), with almost similar blood glucose reduction rate as metformin (30.13±3.75%, p=0.767). Repeated fractionation employing chromatographic separation techniques followed by in vivo anti-hyperglycemic assay yielded partially purified subfractions. A. sessilis ethyl acetate subfraction 4-2 (100mg/kg) displayed remarkable suppression of blood glucose rise in diabetic mice at 6-h post-treatment (26.45±3.75%, p<0.0001), with comparable activity with metformin (100mg/kg, 27.87±5.65%, p=0.652). Liquid chromatography/mass spectrometry showed eight distinct peaks, with four peaks annotated via the Traditional Chinese Medicine library and custom library for A. sessilis. Among these, luteolin, apigenin, ononin, and sophorabioside were identified as putative compounds responsible for the anti-hyperglycemic activity. This result provided basis for the reported anecdotal claims and potential utility of the local variety of A. sessilis leaves as sources of anti-hyperglycemic agents

Preservation of phenolic compounds on dried leaf infusion of Bauhinia forficata Link

Abstract Bauhinia forficata Link aqueous extract is usually recommended as a phytomedicine to reduce blood glucose levels and its biological activity has been linked to the presence of phenolic compounds from B. forficata preparations. Several drying processes are used in the production of dry herbal extracts, which may influence the chemical composition and efficacy of final herbal medicines. Due to significant chemical changes, defining appropriate drying processes is essential for phytopharmaceutical drug development. In view of this, we analyzed dried B. forficata leaf infusion (BFLI) extracts by HPLC-UV-MSn, followed by molecular networking analysis to evaluate the chemical profiles from dried extracts yielded by freeze-and spray-drying processes. The main metabolites detected included 11 ferulic/isoferulic acid derivatives and 13 glycosylated flavonoids. The qualitative chemical profiles were alike for both drying processes, whereas the relative abundance of some flavonoids was higher using spray-drying. Taken together, our results showed that freeze-and spray-drying preserved the phenolic profile of BFLI and suggested that spray-drying may be the most suitable to obtain its dried products. Along with studying the chemical profiles of dried herbal extracts, evaluating the influence of drying processes on the quality and chemical profiles of final products is pivotal and may benefit future research.

Chemical study of Adenocalymma axillarum crude leaf extract and isolated compounds

Abstract Adenocalymma axillarum (K.Schum.) L.G. Lohmann is a liana belonging to the family Bignoniaceae. In traditional medicine, the genus Adenocalymma is used to treat fever, skin ailments, and body, joint, and facial muscle pains, and it is also applied as cosmetic. Biological assays conducted with the A. axillarum crude leaf ethanol extract have indicated leishmanicidal activity and absence of cytotoxicity. This study aimed to analyze the A. axillarum leaf ethanol crude extract by high-performance liquid chromatography-high-resolution mass spectrometry- diode array detector (HPLC-HRMS-DAD) and to evaluate the leishmanicidal and cytotoxic activities of this crude extract, its fractions, and isolated compounds. HPLC-HRMS-DAD analysis of this extract revealed that it consisted mainly of flavonoids, with nine major compounds. Extract purification yielded 4-hydroxy-N-methylproline, 6-β-hydroxyipolamiide, quercetin-3-O-robinobioside, hyperin, isorhamnetin-3-O-robinobioside, and 3'-O-methylhyperin, which were identified by Nuclear Magnetic Resonance. The isolated compounds were inactive against Leishmania amazonensis promastigotes and human lung fibroblast cells.

Leaf extract of Coffea arabica L. reduces lipid peroxidation and has anti-platelet effect in a rat dyslipidemia model

Abstract This study aimed to evaluate the antioxidant potential of the Coffea arabica Lineu (L.) leaf extract and its effects on platelet aggregation of dyslipidemic rats. The extract was obtained by the percolation of C. arabica L. leaves in hydroethanolic solution 70% (v/v). The mass spectrometry FIA-ESI-MS² suggested the presence of chlorogenic acid, rutin acid, and quinic acid. The DPPH• radicals scavenging capacity was demonstrated (IC50 = 0.06 mg/mL). The extract was administered to rats by gavage (300 mg/kg/day) for 56 days. Dyslipidemia was induced by administering Triton WR-1339 (300 mg/kg body weight) on the 54th day. On day 56, blood was collected by puncturing the abdominal aorta artery and the aortic artery was removed. Lipid profile, markers of renal and hepatic injury, lipid peroxidation, and platelet aggregation tests were carried out. The ingestion of extract reduced the lipid peroxidation (aorta and plasma) and platelet aggregation in dyslipidemic rats. The extract did not affect markers of renal and hepatic function as analyzed in this study, suggesting neither impaired liver nor kidney function in these animals. Therefore, our results demonstrate that the extract of leaves of C. arabica L. show antioxidant potential in vitro and in vivo as well as anti-platelet aggregation in dyslipidemic animals

Artichoke extracts with potential application in chemoprevention and inflammatory processes

Abstract The aim of this work is to study three cultivars of artichoke (Cynara cardunculus var. scolymus): Gauchito, Guri and Oro Verde in terms of their in vitro chemoprevention and anti-inflammatory properties. These cultivars show good productive performance. The phenolic composition of their fresh leaves and edible bracts was analyzed by high performance liquid chromatography and high resolution mass spectrometry (HPLC-HRMS), showing mainly caffeoylquinic acids and flavonoids. Caffeoylquinic acids were quantified and the highest content was found in Gauchito cultivar. In this cultivar, the content of dicaffeoylquinic acids in fresh bracts was six times higher than that in fresh leaves (10064.5 ± 378.3 mg/kg versus 1451.0 ± 209.3 mg/kg respectively). Luteolin flavonoids were detected in leaves. The extracts from fresh bracts and leaves were assessed in their in vitro bioactivity against human neuroblastoma cells (SH-SY5Y). Inhibition of SH-SY5Y cells proliferation by Gauchito and Guri leaf extracts (8 µg/mL) was higher than 50 %. The leaf extracts of the same cultivars showed an inhibitory effect on human interferon IFN-I, decreasing its activity 50% at 40 µg/mL. Interestingly, the bract extracts did not show in vitro bioactivity at these concentrations, nor did the pure compounds chlorogenic acid, cynarin, apigenin and luteolin (at 2 µg/mL). These results suggest that Gauchito and Guri leaf extracts have potential for human neuroblastoma chemoprevention and treatment of inflammatory processes.

Anti-hyperlipidemic and anti-diabetic evaluation of ethanolic leaf extract of Catharanthus roseus alone and in combination therapy

The use of plants in disease treatment is cost effective and relatively safe. This study was designed to investigate anti-hyperlipidemic and anti-diabetic activity of ethanolic leaf extract of Catharanthus roseus alone and in combination therapy in hyperlipidemic & diabetic mice. Eight groups comprising five mice each were used. Group A was hyperlipidemic control, group B, C, D received atorvastatin (20 mg/kg), leaf extract (200 mg/kg) and leaf extract in combination with atorvastatin (200 mg/kg and 20 mg/kg) orally for 15 days. Group E was diabetic control. Group F, G, H received sitagliptin (40 mg/kg), leaf extract (200 mg/kg) and extract in combination with sitagliptin (200 mg/kg and 40 mg/kg) orally for 7 days. Blood cholesterol levels were measured at 1st, 5th, 10th and 15th day and fasting blood sugar levels were measured at 2, 12, 24, 72 and 168 hours during treatment. One-way ANOVA with tukey- kramer multiple comparison test was used. The chemical characterization of ethanolic extract of Catharanthus roseus leaves showed presence of alkaloids, saponins, tannins and flavonoids. Ethanolic extract of Catharanthus roseus has significant anti-hyperlipidemic & anti-diabetic effects (p<0.05, p<0.01) when compared with control but had not cause significantly increase in anti-hyperlipidemic effects of atorvastatin. While significantly increased the antidiabetic effect of sitagliptin (p<0.05)

Preliminary prospection of phytotherapic compounds from the essential oils from barks and leaves of Umburana (Commiphora Leptophloeos)

Abstract The potential of the biome caatinga (exclusive from northeastern Brazil) has been evaluated in recent research for application in the pharmaceutical industry. Among the species of medicinal plants from caatinga, one can highlight the Commiphora leptophloeos (umburana), which has been used as infusions and syrups by the regional population for inflammatory and infectious diseases. Essential oils from umburana leaves and barks were obtained in a Clevenger apparatus and analyzed by gas chromatography/mass spectrometry, and total phenolic and flavonoids were determined by spectrophotometric analysis. It was observed that a large part of the major compounds present in the essential oil is described as having antitumor activity, enabling research in investigational oncology with umburana (C. leptophloeos). In addition, some little explored components have been identified, such as cadinene, alpha-selinene, and elemenone. Despite being easily found in several plants, there are no clinical trials involving their biological activity in a well-defined isolated form, which could make exploring new studies possible. Furthermore, the presence of phenolic compounds and flavonoids allows future studies about the potential antimicrobial and antioxidant activity.

Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from Lippia alba

Abstract The composition and pharmacological properties of Lippia alba (Mill.) (L. alba) (Verbenaceae) flower and leaf essential oils (EO) were determined in this study. The major constituents in the flower EO were geranial (49.83%) and neral (32.75%), and in the leaf EO were geranial (38.06%), neral (31.02%), and limonene (18.03%). Flower EO inhibited thrombolysis induced by Bothrops moojeni (B. moojeni) and Lachesis muta muta (L. muta muta) venoms (0.05-1.2 µL mL-1). When tested against L. muta muta venom, the protective effect was smaller in both EO. The EOs prolonged the clotting time induced by L. muta muta venom and a procoagulant effect was observed on B. moojeni. In the comet assay, the flower EO presented anti-genotoxic action (damage frequency of only 11.6 - 34.9%) against the L. muta muta venom. The positive control (Doxorubicin) and the venom alone presented a damage frequency of 80.3% and 70.7%, respectively. The flower EO protected DNA from damage induced by L. muta muta venom. L. alba leaf and flower EOs presented anti-genotoxic action

Assessing compatibility of excipients selected for a sustained release formulation of bilberry leaf extract

Abstract The study is aimed to assess the compatibility of bilberry leaf powder extract (BLPE) with six excipients selected for sustained-release (SR) tablet formulation. The BLPE was obtained with the addition of L-arginine and Myo-inositol as the carriers. Thermogravimetric (TG-DTG) analysis and Fourier-transform infrared spectroscopy (FTIR), supported by Pearson correlation analysis, were applied to detect possible interactions in the binary mixtures (1:1) of the BLPE with each excipient. The TG-DTG showed some deviations in the thermal behavior of the BLPE / excipient mixtures. However, only the thermal behavior of magnesium stearate in the mixture significantly differed from individual samples, which suggested chemical interaction for this excipient. The FTIR analysis confirmed that the BLPE is compatible with Eudragit L100, Methocel K4M, Methocel K100LV, Avicel PH-101, and Plasdone S-630. Whereas it undergoes solid-state chemical interaction in the binary mixture with magnesium stearate. According to the FTIR-spectra, it is suggested that this interaction results in the formation of stearic acid and alkalization of the medium. These findings evidence for the possibility of using TG-DTG analysis as an independent thermal technique for compatibility studies and also confirm the earlier reported interaction of basic lubricants, e.g., stearic salts, with active ingredients containing amino groups.

Antidepressant-like activity of ethanol extract of leaves of Caesalpinia pulcherrima in unstressed and stressed mice

In the present study, antidepressant-like activity of ethanol extract of leaves of Caesalpinia pulcherrima was evaluated in Swiss young male albino mice. Stress was induced in mice by subjecting them to unpredictable mild stress for 21 successive days. Ethanol extract of the leaves (100, 200 and 400 mg/ kg, p.o.) and fluoxetine (20 mg/kg, p.o.) were administered for 21 consecutive days to separate groups of unstressed and stressed mice. Ethanol extract (200 and 400 mg/kg) and fluoxetine significantly decreased immobility period of unstressed as well as stressed mice in tail suspension test (TST). However, the lowest dose (100 mg/kg) of the extract also significantly decreased immobility period of stressed mice in TST. The extract significantly restored reduced sucrose preference in stressed mice. There was no significant effect on locomotor activity of mice. Ethanol extract of the leaves significantly decreased plasma nitrite and corticosterone levels; brain MAO-A activity and MDA level; and increased brain reduced glutathione and catalase activity in unstressed as well as stressed mice as compared to their respective vehicle treated controls. Thus, ethanol extract of leaves of Caesalpinia pulcherrima showed significant antidepressant-like activity in unstressed and stressed mice probably through inhibition of brain MAO-Aactivity, reduction of oxidative stress and plasma corticosterone levels.

Antibacterial substances from leaves of Protium spruceanum (Burseraceae): in vitro and in silico evaluation

Due to the increase of bacterial resistance, the search for new antibiotics is necessary and the medicinal plants represent its most important source. The aim of this study was to evaluate the antibacterial property of extract and fractions from Protium spruceanum leaves, against pathogenic bacteria. By means of diffusion and microdilution assays, the crude extract was active against the nine bacteria tested being the hydromethanolic fraction the most active. During phytochemical procedures, procyanidin (1) and catechin (2) were identified as the main antibacterial constituents of this fraction. In silico results obtained using PASSonline tool indicated 1 and 2 as having good potential to interact with different targets of currently used antibiotics. These results no indicated potential to none DNA effect and indicated the cell wall as mainly target. Electrophoresis result supported that had no DNA damage. Cell wall damage was confirmed by propidium iodide test that showed increased membrane permeability and by cell surface deformations observed in scanning electronic microscopy. The in vitro assays together with the in silico prediction results establish the potential of P. spruceanum as source of antibacterial compounds that acts on important bacterial targets. These results contribute to the development of natural substances against pathogenic bacteria and to discovery of new antibiotics.

Feeding preference of the shredder Phylloicus sp. for plant leaves of Chrysophyllum oliviforme or Miconia chartacea after conditioning in streams from different biomes / Preferência alimentar do fragmentador Phylloicus sp. por folhas de Chrysophyllum oliviforme e Miconia chartacea após condicionadas em riachos de dois biomas

Abstract Macroinvertebrate shredders consume preferably leaves conditioned by fungi and bacteria which offer greater palatability to them. Plant species in Cerrado present high concentration of chemical elements such as lignin and cellulose, phenols and tanins thus making them less attractive for shredders consumption and limiting the palatability. This study aimed to evaluate the feeding preference of a macroinvertebrate shredder of the genus Phylloicus for plant material from two different biomes (Cerrado and Mata Atlântica), after conditioning in a stream of Mata Atlântica and observing their physical and chemical characteristics. Senescent leaves were collected, monthly from the litterfall of riparian vegetation in a 500 m stretch of a stream in each biome from August 2014 to January 201. The most abundant species in each stream was selected for the experiment. The experimental design consisted in with two treatments. The first (T1) comprised leaf discs from Chrysophyllum oliviforme (Cerrado species) together with leaf discs of Miconia chartacea (Atlantic Forest species) which were conditioned in the Atlantic Forest stream. The second treatment (T2) involved leaf discs of Miconia chartacea conditioned in Mata Altlântica and Cerrado streams. Both tests had showed significant differences between the two treatments (T1 and T2). For T1, there was consumption of M. chartacea leaf discs by Phylloicus sp., but there was no consumption of C. oliviforme discs. For T2, there was preference for M. chartaceae leaves conditioned in a stream of Mata Atlântica than in Cerrado stream. The results showed that Phylloicus sp., had presented preference for food detritus of the Mata Antlântica biome and rejection to the one from Cerrado biome.

Resumo Macroinvertebrados fragmentadores consomem folhas preferencialmente condicionadas por fungos e bactérias que lhes oferece uma maior palatabilidade. Nas espécies do cerrado esse condicionamento está também associado às altas concentrações de elementos químicos limitantes à palatabilidade como alto teor de lignina e celulose, que tornam as folhas menos atrativas para os fragmentadores. O trabalho teve como objetivo avaliar a preferência alimentar de macroinvertebrados fragmentador ( Phylloicus sp.) por material vegetal de dois diferentes biomas (Cerrado e Mata Atlântica), após condicionamento em riacho de Mata Atlântica, observando suas características físicas e químicas. Foram coletadas folhas senescentes do aporte vegetal (AV) de espécies nativas da vegetação ripária nesses dois biomas, com periodicidade mensal em um trecho de 500 m de um córrego em cada bioma. O experimento foi delineado com dois tratamentos. O primeiro (T1) compreendeu discos de folhas do Cerrado (Chrysophyllum oliviforme) mais discos de folhas da Mata Atlântica (Miconia chartacea) que foram condicionadas em córrego de Mata Atlântica. O segundo tratamento (T2) envolveu discos de folhas da Mata Atlântica condicionadas em córrego da Mata Atlântica mais discos de folhas da Mata Atlântica condicionadas em córrego do Cerrado. Os dois testes apontaram diferenças significativas entre os dois tratamentos (T1 e T2). Para T1 houve consumo de discos de folha de M. chartacea por Phylloicus sp, mas não houve consumo dos discos de C. oliviforme , de Cerrado. Para T2, houve o consumo, porém a preferência pelas folhas de M. chartaceae condicionadas no córrego da Mata Atlantica foi consideravelmete maior. Os resultados apontam que Phylloicus sp, apresentou preferência alimentar pelo detrito de bioma de Mata Atlântica e rejeição pelo detrito do bioma Cerrado.

Antileishmanial in vitro activity of essential oil from Myrciaria plinioides, a native species from Southern Brazil

In South American folk medicine members of the genus Myrciaria are used for the treatment of malaria, diarrhoea, asthma, inflammation and post-partum uterine cleansing. The aim of this work was to evaluate its antileishmanial properties (in vitro) of essential oil derived from leaves of Myrciaria plinioides D. Legrand, a plant species that is native in South of Brazil. The essential oil was obtained by hydro-distillation using fresh leaves of M. plinioides. The chemical composition of this essential oil (MPEO, M. plinioides essential oil) was determined by gas chromatography coupled to mass spectrometry (GC-MS). MPEO was assayed in vitro for antileishmanial properties against promastigotes of Leishmania amazonensis and Leishmania infantum, and for cytotoxicity against murine peritoneal macrophages. The MPEO comprised 66 components and was rich in oxygenated sesquiterpenes (82.66%) containing spathulenol (21.12%) as its major constituent. The MPEO was effective against L. amazonensis with IC50 value of 14.16 ± 7.40 µg/mL, while against L. infantum the IC50 value was higher with 101.50 ± 5.78 µg/mL. The MPEO showed significant activity against L. amazonensis, and presented a selectivity index (SI) of 6.60. The results suggest that the essential oil from leaves of M. plinioides is a promising source for new antileishmanial agents against L. amazonensis.

Ocotea nutans (Nees) Mez (Lauraceae): chemical composition, antioxidant capacity and biological properties of essential oil

The present study was undertaken to assess the potential uses of the essential oil obtained from Ocotea nutans (Nees) Mez. The hydrodistilled essential oil from O. nutans leaves was analyzed by gas chromatography-mass spectrometry. Fifty-eight compounds representing 87,29% of the total leaf essential oil components were identified, of which biciclogermacrene(11.41%), germacrene-D (4.89%), bisabolol-11-ol(3.73%) and spathulenol (3.71%) were the major compounds. The essential oil from O. nutans were tested for antibacterial activity using the minimum inhibitory concentracion (MIC) method, Artemia salina method, larvicidal activity in Aedes aegypti, and antioxidant capacity.The antioxidant activity measured by the phosphomolybdenum complex and Prussian blue method had positive results. The minimum inhibitory concentration for the microorganisms tested allowed moderate inhibitionfor Enterococcus faecalis (MIC=500 µg/mL). Artemia salina were toxic to the organisms in the study (LC50= 71,70 µg /mL). The essential oil showed remarkable larvicidal activity potencial (LC50= 250 µg/mL). The present results showed thatO. nutans essential oil has potential biological uses

Investigations of Acacia modesta Wall. leaves for in vitro anti-diabetic, proliferative and cytotoxic effects

ABSTRACT The leaves of Acacia modesta Wall. have been shown to possess diverse pharmacological properties. Therefore, we aimed at evaluating anti-diabetic, cytotoxic and proliferative effects of extracts of Acacia modesta Wall. leaves. After evaluating the primary and secondary metabolites, anti-diabetic activity of various extracts was assessed by α-amylase inhibition, glucose uptake by yeast cells and non-enzymatic glycosylation of hemoglobin assay. Cytotoxicity and proliferative potential was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and short term proliferation assays, respectively, using human liver carcinoma cell line, HepG2. Among other extracts, chloroform extract exhibited 34.16% inhibition of α-amylase, 90.65% inhibition of hemoglobin glycosylation and 94.75% glucose uptake employing α-amylase inhibition, non-enzymatic glycosylation of hemoglobin and glucose uptake by yeast cells assays, respectively. Moreover, extracts exhibited no significant effects on HepG2 cell viability and proliferation. So, this data suggested that chloroform extract of leaves of Acacia modesta Wall., exhibited higher anti-hyperglycemic activity in comparison to extracts in other solvents, while no extract demonstrated cytotoxic and proliferation effects when tested using HepG2 cell line

Determination of phytoconstituents of n-hexane extract of leaves of Morus nigra and evaluation of their effects on biochemical and histopathological parameters in paracetamol intoxicated mice liver

In-vitro investigation of Morus nigra L. (Moraceae) has demonstrated the evidence of several antioxidant compounds. Current study was aimed to determine the phytoconstituents and hepatoprotective potential of n-hexane extract of Morus nigra. Swiss albino mice were divided into five groups: normal control, paracetamol control, silymarin control, 250 mg/kg p.o. extract for 7 days plus paracetamol 3 h later, 500 mg/kg p.o. extract for 7 days plus paracetamol 3 h later. Serum levels of liver enzymes and total bilirubin were assessed and compared between the groups by using one-way ANOVA to confirm hepatoprotective activity. HPLC analysis revealed the presence quercetin, oleanolic acid, luteolin, apigenin, vitamin C and kuwanon C. n-hexane extract of Morus nigra at dose of 250 mg/kg reduced elevated ALT by 54.3% (p<0.001), AST by 55.1% (p<0.01), ALP by 28.5% (p<0.01) and TBR by 56.4% (p<0.01) as compared to paracetamol control. The dose of 500 mg/kg of the extract reduced the ALT levels by 62.9% (p<0.001), AST by 56.7% (p<0.01), ALP by 33.6% (p<0.01) and TBR by 54.5% (p<0.01), as compared to paracetamol group. However, the reduction of liver enzymes and total bilirubin after administration of extract was comparable to the silymarin. Current study demonstrated potential hepatoprotective activity of n-hexane extract of Morus nigra

The utility of Bambusoideae (Poaceae, Poales) leaf blade anatomy for identification and systematics / A utilidade da anatomia da lâmina foliar para a identificação e sistemática de espécies de Bambuboideae (Poaceae, Poales)

Abstract Bambusoideae is a diverse subfamily that includes herbaceous (Olyreae) and woody (Arundinarieae and Bambuseae) bamboos. Species within Bambusae are particularly difficult to identify due to their monocarpic lifecycle and the often long durations between mass flowering events; whereas the herbaceous bamboos are pluricarpic, but often are found with no reproductive structures. The leaf blade anatomy of 16 sympatric species of native Brazilian bamboos (Olyreae and Bambuseae) from the Atlantic Rainforest was studied in order to detect useful features for their identification. All the studied species share the following features: epidermis with a single stratum of cells; adaxial bulliform cells; mesophyll with arm cells, rosette cells, and fusoid cells; and collateral vascular bundles. Herbaceous bamboos share two features: papillae scattered on the abaxial surface and parallel-sided arrays of bulliform cells; whereas woody bamboos share: centrally organized papillae and fan-shaped arrays of bulliform cells. Also within the woody bamboos, intercostal fibers and a midrib with only one vascular bundle (simple midrib) characterize the subtribe Arthrostylidiinae; whereas a midrib with more than one vascular bundle (complex midrib) and a stomatal apparatus with two pappilae per subsidiary cell characterize the subtribe Chusqueinae. There are also diagnostic features for the sampled species, such as: papillae shape, and the outline and structure of the midrib. An identification key for all the studied species is provided based on the anatomical features.

Resumo Bambusoideae é uma subfamília que inclui diversas espécies de bambus herbáceos (Olyreae) e lignificados (Arundinarie e Bambuseae). Bambus lignificados geralmente apresentam dificuldades de delimitação e identificação, devido principalmente ao ciclo monocárpico e longa amplitude temporal entre florações; enquanto que bambus herbáceos possuem ciclo pluricárpico, porém frequentemente são encontrados em estágio vegetativo. Foi estudada a anatomia da lâmina foliar de 16 espécies de Bambusoideae (Olyreae e Bambuseae), simpátricas e nativas do Brasil, visando levantar caracteres úteis para sua identificação. Todos os táxons estudados compartilham: epiderme uniestratificada; células buliformes na face adaxial; mesofilo com células invaginantes, células em roseta e células fusoides; e feixes vasculares colaterais. Bambus herbáceos compartilham: papilas dispersas na face abaxial e grupos de células buliformes organizadas paralelamente; enquanto que bambos lignificados compartilham: papilas organizadas em colunas centrais e grupos de células buliformes em forma de leque. Ainda dentre os bambus lignificados, fibras intercostais e nervura central com apenas um feixe vascular (nervura central simples) caracterizam a subtribo Arthrostylidiinae; enquanto que nervura central com mais de um feixe vascular (nervura central complexa) e aparelho estomático com duas papilas por célula subsidiária caracterizam a subtribo Chusqueinae. Há ainda caracteres anatômicos diagnósticos, tais como: forma da papila, e forma e estrutura da nervura central. Uma chave de identificação é fornecida baseada nos caracteres anatômicos relevantes à identificação das espécies estudadas.

Análise farmacobotânica de folha e caule de Tanacetum vulgare (L. ) / Pharmacobotanical analysis of leaf and stem of Tanacetum vulgare (L. )

RESUMO Tanacetum vulgare L., conhecida popularmente como catinga-de-mulata, é utilizada na medicina tradicional como vermífugo, digestivo e emenagogo. Objetivou-se analisar a morfoanatomia de folhas e caules dessa espécie medicinal com vistas ao controle da qualidade. Foram utilizadas técnicas usuais de microscopia de luz e eletrônica de varredura. As folhas são compostas, pinatissectas e anfiestomáticas. Tricomas tectores flageliformes simples e glandulares capitados são observados. A nervura central tem formato biconvexo, o pecíolo é côncavo-convexo, a ráque é plano-convexa e o caule é arredondado. Feixes vasculares colaterais, calotas de fibras perivasculares e colênquima lamelar estão presentes na folha e no caule. Os caracteres morfoanatômicos evidenciados contribuem na identificação do táxon e fornecem subsídios farmacobotânicos para o controle da qualidade de drogas vegetais e fitoterápicos.

ABSTRACT Tanacetum vulgare L., popularly known as tansy, is used in traditional medicine as a vermifuge, digestive and emmenagogue. This study aimed to analyze the morphoanatomical leaves and stems of this medicinal plant in order to control the quality. Usual techniques of light and scanning electron microscopy were used. The leaves are composed, pinatissect and amphistomatic. Simple and flagelliform non- glandular trichomes and capitate glandular trichomes are observed. The midrib has biconvex shape, the petiole is concavo-convex, the rachis is plano-convex, and the stem is rounded. Collateral vascular bundles, perivascular fiber caps and laminar collenchyma are encountered in the leaves and stems. Evidenced the morphological and anatomical features contribute to the identification of the taxon and provide pharmacobotanical data for the quality control of herbal drugs.

Fenologia e prospecção fitoquímica do jaborandi (Pilocarpus microphyllus Stapf ex Holmes) / Phenology and phytochemical prospection of Jaborandi (Pilocarpus microphyllus Stapf ex Holmes)

RESUMO O estudo fenológico tem como finalidade determinar o ritmo sazonal dos eventos do ciclo de vida da planta, como floração e frutificação. Estes eventos são determinados por uma série de fatores, como: alternância de períodos chuvosos ou não chuvosos, intensidade da radiação solar, entre outros. A fitoquímica tem por objetivos conhecer os constituintes químicos de espécies vegetais ou avaliar sua presença. O presente trabalho teve como objetivo realizar a caracterização fenológica e a prospecção fitoquímica de folhas de jaborandi. A área de estudo para a avaliação do material vegetal foi o Banco Ativo de Germoplasma de Jaborandi da Embrapa Amazônia Oriental, situada no município de Belém-PA. Os acessos escolhidos foram: Merck, cultivado a pleno sol e à sombra; Japonês e Bonal 4, cultivados a pleno sol. Os registros foram realizados diariamente por um período de 28 meses correspondendo a agosto de 2010 a dezembro de 2012, de cinco plantas/acesso e organizados para demonstração mensal, através de fichas com a numeração respectiva das plantas, com registro de presença ou ausência das fenofases, floração e frutificação. A determinação do peso seco das amostras coletadas dos acessos foi realizada no Laboratório de Agroindústria da Embrapa Amazônia Oriental, onde, após a triagem e remoção das impurezas, as folhas foram cortadas, pesadas, colocadas em bandejas de inox e secas em estufa com circulação mecânica (FANEM 320-SE), à temperatura de 45º C por 120 h. Em seguida, as amostras foram pesadas, trituradas e acondicionadas em sacos plásticos devidamente identificados e guardados sob refrigeração à temperatura de 10º C até o uso. Os extratos das plantas foram preparados utilizando-se 100 g de folhas secas de cada acesso, triturados e submetidos à extração hidroalcoólica (etanol 80%) em banho-maria sob refluxo, por aproximadamente 4 horas. Os extratos foram armazenados protegidos da luz na geladeira até o momento das análises. Foi analisada a presença das seguintes classes de substâncias químicas: ácidos orgânicos, açúcares redutores, polissacarídeos, proteínas e aminoácidos, taninos, catequinas, flavonoides, glicosídeos cardíacos, lactonas sesquiterpênicas, azulenos, carotenoides, esteroides e triterpenoides, depsídeos e depsidonas, derivados da cumarina, saponina espumídica, alcaloides, purinas, antraquinonas. Os resultados obtidos demonstraram que a espécie Pilocarpus microphyllus apresentou floração durante o ano todo e frutificação em onze meses, e a prospecção fitoquímica revelou a presença de 11 classes de constituintes químicos.

ABSTRACT Phenological studies aim to determine the seasonal rhythm of the plant life cycle events, as flowering and fruiting. These events are determined by different factors, such as: alternating periods of rainy or dryer seasons, solar radiation intensity, among others. Phytochemistry aims to identify the chemical constituents of plant species or to evaluate their presence. This study aimed the phenological characterization and the phytochemical prospection of jaborandi leaves. The chosen study area for the plant material assessment was the Active Germplasm Bank of Jaborandi in the Embrapa Eastern Amazon, located in the city of Belém, PA, Brazil. The chosen accessions were the following: Merck, grown in full sun and in shade; Japanese and Bonal 4, both grown in full sun. Records were taken on a daily basis for a period of 28 months (August of 2010 to December of 2012), from five plants/accession, and arranged for monthly demonstrations by record sheets containing the corresponding plant numeration and the presence or absence of flowering and fruiting phenophases. The dry weight of the samples collected from the accessions was measured at the Laboratory of Agribusiness of Embrapa Eastern Amazon, where, after the sorting and removal of impurities, the leaves were cut, weighed, placed in stainless steel trays, and dried in forced air circulation oven (FANEM 320 UP) at a 45°C for 120h. Then, the samples were weighed, crushed, and placed in plastic bags properly identified and stored under refrigeration at a temperature of 10ºC until the use. The plant extracts were prepared using 100g of dried leaves from each accession, crushed, and subjected to hydroalcoholic extraction (80% ethanol) with water bath heating under reflux for approximately 4 hours. The extracts were stored protected from light in a refrigerator until the analysis. We analyzed the presence of the following classes of chemical substances: organic acids, reducing sugars, polysaccharides, proteins and amino acids, tannins, catechins, flavonoids, cardiac glycosides, sesquiterpene lactones, azulenes, carotenoids, steroids and triterpernoids, depsides and depsidones, coumarin derivatives, foam saponin, alkaloids, purines, anthraquinones. Our results showed that the flowering of Pilocarpus microphyllus occurred throughout the year and fruiting occurred in eleven months, and the phytochemical prospection revealed the presence of 11 classes of chemical constituents.

Characterization saprobic fungi on leaf litter of two species of trees in the Atlantic Forest, Brazil

Abstract We investigated the composition and structure of fungal communities associated with leaf litter generated by Clusia nemorosa and Vismia guianensis that belong to phylogenetically-related botanical families and exist together in a remnant of the Atlantic Forest in Bahia, Brazil. Samplings were conducted during wet (June 2011) and dry (January 2013) seasons in Serra da Jibóia. The fungi were isolated using particle filtration and the 1,832 isolates represented 92 taxa. The wet season yielded the largest number of isolates (1,141) and taxa (76) compared with the dry season (641 isolates and 37 taxa). The richness and diversity of fungal species associated with C. nemorosa (64 taxa, Simpson=0.95)were higher compared with those of V.guianensis (59 taxa, Simpson =0.90). Analysis of similarity (ANOSIM) revealed significant variations in the composition and community structure of fungi isolated from the two plants as a function of seasons. In contrast, nonmetric multidimensional scaling (NMDS) analysis show that the seasonality was an important influence on the distribution of fungal species. However, the populations of the saprobic fungal communities were dynamic, and several factors may influence such communities in the Atlantic Forest.