Erysothrine, an alkaloid extracted from flowers of Erythrina mulungu Mart. ex Benth: Evaluating its anticonvulsant and anxiolytic potential

In this study, we isolated the alkaloid erysothrine from the hydroalcoholic extract of flowers from E. mulungu and screened for its anticonvulsant and anxiolytic actions based on neuroethological and neurochemical experiments. Our results showed that the administration of erysothrine inhibited seizures evoked by bicuculline, PTZ, NMDA and most remarkably, kainic acid. Also, erysothrine induced an increase in the number of entries but not in the time spent in the open arms of the EPM. However, we did not notice any alterations in the light-dark choice or in the open-field tests. In preliminary neurochemistry tests, we also showed that erysothrine (0.001-10 mu g/mL) did not alter the GABA or glutamate synaptossomal uptake and binding. Altogether, our results describe an alkaloid with anticonvulsant activity and mild anxiolytic activity that might be considered well tolerated as it does not alter the general behavior of the animals in the used doses. (C) 2012 Elsevier Inc. All rights reserved.

Aplicação de espectrometria de massas com ionização por elétron na análise de alcaloides do mulungu

Erythrina verna is a medicinal plant used to calm agitation popularly known as mulungu. We purchased the barks of E. verna from a commercial producer and analyzed the alkaloid fraction of the bark by CG-MS and HRESI-MS. Five erythrinian alkaloids were identified: erysotrine, erythratidine, erythratidinone, epimer, and 11-hydroxieritratidinone. Here we report the compound 11-hydroxieritratidinone for the first time as a natural product.

Aplicação de espectrometria de massas com ionização por elétron na análise de alcaloides do mulungu

Erythrina verna is a medicinal plant used to calm agitation popularly known as mulungu. We purchased the barks of E. verna from a commercial producer and analyzed the alkaloid fraction of the bark by CG-MS and HRESI-MS. Five erythrinian alkaloids were identified: erysotrine, erythratidine, erythratidinone, epimer, and 11-hydroxieritratidinone. Here we report the compound 11-hydroxieritratidinone for the first time as a natural product.

Reserve carbohydrates and lipids from the seeds of four tropical tree species with different sensitivity to desiccation

Considering the importance of water content for the conservation and storage of seeds, and the involvement of soluble carbohydrates and lipids for embryo development, a comparative study was carried out among the seeds of Inga vera (ingá), Eugenia uniflora (pitanga), both classified as recalcitrant, and Caesalpinia echinata (brazilwood) and Erythrina speciosa (mulungu), considered as orthodox seeds. Low concentrations of cyclitols (0.3-0.5%), raffinose family oligosaccharides (ca. 0.05%) and unsaturated fatty acids (0-19%) were found in the seeds of ingá and pitanga, while larger amounts of cyclitols (2-3%) and raffinose (4.6-13%) were found in brazilwood and mulungu, respectively. These results, in addition to higher proportions of unsaturated fatty acids (53-71%) in orthodox seeds, suggested that sugars and lipids played important role in water movement, protecting the embryo cell membranes against injuries during dehydration.

Seropositivity to anti-phenolic glycolipid-I in leprosy cases, contacts and no known contacts of leprosy in an endemic and a non-endemic area in northeast Brazil

The seroprevalence rates of IgM anti-phenolic glycolipid-I (PGL-I) antibodies in four study groups with differing exposure to Mycobacterium leprae in Ceara. Brazil were investigated between March 2005 and August 2006. The first three groups in a high prevalence area included 144 cases of leprosy, their 380 contacts and 317 participants with no known leprosy contact. The fourth group in a low prevalence area consisted of 87 participants with no known leprosy contact living in an area in which no cases of leprosy had been reported in the previous 6 months. Seropositivity and levels of IgM antibodies to PGL-I were investigated using ELISA. The seropositivity levels of anti-PGL-I among the different clinical forms of leprosy cases were 61% for lepromatous, 25% for tuberculoid and 27% indeterminate. The levels of anti-PGL-I antibodies in the endemic area differentiated leprosy cases from non-cases. However, the seropositivity was similar among contact cases (15.8%) and no known leprosy contact cases from high (15.1%) and low (13.8%) prevalence areas. The seropositivity of both contacts and no known contacts was much higher than previously reported among no known contacts in other endemic areas. The study indicates that anti-PGL-I antibodies are not useful as immunological markers of household leprosy contacts and no known leprosy contacts in endemic areas. (C) 2010 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.

Resposta morfogênica de embriões zigóticos de Erythrina velutina Willd: (Leguminosae) cultivados in vitro

O cultivo in vitro de embriões zigóticos é uma técnica promissora para se avançar no estudo do desenvolvimento embrionário e da quebra da dormência de sementes. Diante do exposto, objetivou-se, com este trabalho, avaliar o efeito dos reguladores vegetais 6-benzilaminopurina (BAP) e ácido naftalenoacético (ANA) no potencial morfogenético, in vitro, de embriões zigóticos de mulungu. Embriões zigóticos maduros, oriundos de sementes foram utilizados inteiros, ou seccionados em plúmula, região intermediária e radícula, sendo posteriormente inoculados em meio de cultura WPM, suplementado com combinações de BAP (0,0; 2,0; 4,0; 8,0; 12,0 e 16,0 µM) e ANA (0,0; 1,0 e 2,0 µM), acrescido de 87,64 mM de sacarose e solidificado com 0,7% de ágar. Após 30 dias, avaliaram-se a percentagem de regeneração dos embriões e ápice plumular, o número de brotos, o número de folhas, o comprimento da parte aérea dos brotos, o número de raízes e a percentagem de formação de calos oriundos da região intermediária e da radícula. É possível a regeneração in vitro de mulungu, a partir dos explantes plúmula e embriões zigóticos inteiros, cultivados em meio de cultura WPM, suplementado com 4,0 µM de BAP. Regiões intermediárias e da radícula promoveram a formação de calos compactos (96,06%), na combinação de 10,63 µM BAP e 2,0 µM de ANA.

Armazenamento de sementes de Erythrina velutina willd

O trabalho teve como objetivo avaliar o efeito de diferentes embalagens e ambientes de armazenamento na manutenção da qualidade fisiológica de sementes de Erythrina velutina Willd.. O experimento foi conduzido no Laboratório de Análise de Sementes do Departamento de Fitotecnia do Centro de Ciências Agrárias da Universidade Federal da Paraíba (CCA-UFPB), localizado no município de Areia (PB). As sementes de Erythrina velutina Willd. foram coletadas embaixo de árvores matrizes, no mesmo município e levadas para o laboratório, onde se realizou a homogeneização e acondicionamento em embalagens de papel, pano e vidro e, posterior armazenamento em condições não controladas de laboratório (±25ºC), geladeira (6 ± 2ºC) e câmara fria (4 ± 2ºC), por um período de 225 dias. Antes e após os intervalos de 45 dias foram retiradas amostras de cada embalagem e ambiente de armazenamento para avaliação das seguintes características: teor de água, emergência (porcentagem e índice de velocidade), comprimento e massa seca de epicótilo, hipocótilo e raiz das plântulas normais. O delineamento experimental utilizado foi inteiramente ao acaso com quatro repetições de 25 sementes para cada teste. As sementes de mulungu são ortodoxas e acondicionando-as nas embalagens de papel, pano ou vidro podem ser armazenadas nos ambientes de laboratório, geladeira e câmara fria, durante 225 dias sem perdas significativas na emergência das plântulas.

Indicadores de sustentabilidade para conservação genética de Erythrina velutina Willd., em área de mata ciliar

Um dos grandes desafios para o sistema agropecuário é estabelecer a produção agrícola aliada à conservação dos recursos genéticos, principalmente visando à prote��ão de Áreas de Preservação Permanente. Nesse contexto, mulungu (Erythrina velutina Willd), entre outras espécies nativas, vem sofrendo com as pressões antrópicas nos mais diversos ecossistemas, o que causa reduções em sua base genética. Este trabalho foi realizado com o objetivo de utilizar parâmetros ecológicos e genéticos populacionais como indicadores de sustentabilidade em duas populações naturais de mulungu, situadas em áreas de floresta ciliar, no Estado de Sergipe, bem como avaliar a tendência à sustentabilidade delas, visando à conservação genética da espécie. Utilizou-se a matriz de Pressão-Estado-Impacto/Efeito-Resposta (PEI/ER) com a sele��ão de 13 indicadores, a partir do uso de marcadores moleculares RAPD e bioquímicos (enzimas) nas populações, de modo a apresentá-los como informações relevantes para medir o progresso quanto à sustentabilidade e conservação de mulungu. As populações estudadas apresentaram baixa tendência à sustentabilidade, necessitando de estratégias para mudança desse status.

Linfadenite caseosa em ovinos deslanados abatidos em um frigorífico da Paraíba

Este trabalho teve como objetivo determinar a prevalência e distribuição de lesões da linfadenite caseosa em ovinos deslanados abatidos no município de Mulungu, Paraíba. Dos 1466 ovinos abatidos, 236 (15,9%) apresentaram lesões macroscópicas semelhantes à linfadenite caseosa. A prevalência foi maior em fêmeas, 17.9% (135/754) que em machos, 13,8% (101/732). As principais lesões estavam localizadas nos linfonodos pré-escapulares em 36,3% (97/268) dos ovinos, nos parotídeos em 22,4% (60/268) e no pré-crural em 20,9% (56/268). Somente 70 (26,1%) de um total de 268 lesões foram detectadas no exame ante mortem. Foram encontradas lesões em todos os lotes provenientes de diversos municípios da Paraíba, Pernambuco e Bahia. Das 51 amostras em que foi feito o cultivo bacteriológico, em 74,5% (43/51) foi isolado Corynebacterium pseudotuberculosis. Em 7,8% (4/51) foi isolado Staphylococcus aureus, em 2% (1/51) Escherichia coli e 5,9% (3/51) amostras foram negativas. No exame histológico, em 11 linfonodos havia lesões da linfadenite caseosa caracterizadas por área de necrose central formada por lamelas concêntricas, com presença de grandes colônias bacterianas e focos de mineralização, rodeada por uma faixa de infiltrado inflamatório com macrófagos epitelióides e poucos neutrófilos. Na camada adjacente observaram-se linfócitos e plasmócitos e toda a lesão era delimitada por tecido conjuntivo fibroso. Nas outras 32 amostras foram observadas a maioria, mas não todas as lesões características da doença. Em 21 amostras foram encontradas células gigantes. As lesões histológicas dos linfonodos que foi isolado S. aureus foram semelhantes as da linfadenite caseosa. Conclui-se que as lesões histológicas de linfadenite caseosa são características, mas não patognomônicas, pois podem ser confundidas com lesões causadas por outros organismos piógenos e com tuberculose; portanto o isolamento bacteriológico é imprescindível para o diagnóstico definitivo da linfadenite caseosa.

Morfologia de frutos, sementes, plântulas e plantas de Erythrina velutina willd., leguminoseae - Papilionideae

O conhecimento dos aspectos morfológicos de sementes e plântulas pode ser usado em estudos de taxonomia, na interpretação de testes de germinação em laboratório, trabalhos em viveiro e em estudos relacionados à ecologia da espécie. Os objetivos deste trabalho foram o de descrever e ilustrar a morfologia externa e interna dos frutos e sementes, as fases de germinação, a plântula e as plantas de mulungu (Erythrina velutina Willd.). Os frutos e sementes foram coletados no solo, embaixo de árvores matrizes distribuídas em diferentes fragmentos florestais, no município de Areia - PB. Para a descrição morfológica dos frutos, sementes, germinação e desenvolvimento da plântula foram examinados cem unidades que foram selecionadas aleatoriamente. A semente é reniforme; embrião axial com cotilédones carnosos; germinação epígea fanerocotiledonar e na fase de plântula foi observado heterofilia. Os aspectos descritos e ilustrados mostraram-se homogêneos e confiáveis para a identificação da espécie. Deste modo, podem ser úteis em trabalhos de tecnologia de sementes, para a avaliação qualidade fisiológica de sementes e avaliação de plântulas em testes de germinação, além de servirem para identificação da espécie em viveiros e também para estudos de regeneração natural.

Mobilization of reserves and germination of seeds of Erythrina velutina Willd. (Leguminosae - Papilionoideae) under different osmotic potentials

Some environmental factors, including water availability, may influence seed germination. This study investigated the germination of E. velutina seeds submitted to different osmotic potentials and mobilization of reserves during water-stress. Scarified seeds were arranged in paper rolls and soaked in solutions of Polyethylene Glycol (PEG) prepared in osmotic potentials 0.0, -0.2, -0.4, -0.6, and -0.8 MPa and kept into a seed germinator, at 25 °C, and 12/12 h photoperiod (L/D), during 10 days. The percentage, mean time, mean speed, germination speed index; as well as the germination uniformity coefficient were assessed. During germination process the total soluble sugars, reducing sugars, soluble protein, and total amino acids were quantified in the cotyledon, hypocotyl and radicle of soaked seeds and cotyledons of quiescent seeds (control). There was influence of osmotic potential on E. velutina seed germination. The germination percentage remained at high levels until -0.6 MPa and above this osmotic potential there has been no germination. The mobilization of stored reserves of carbon and nitrogen in E. velutina seeds was also influenced by water-stress. There was sensitiveness between -0.2 and -0.6 MPa; however, the degradation and the mobilization of reserves was slower when the osmotic potential decreased.

Avaliação das atividades anti-inflamatória, anticoagulante e antiproliferativa do inibidor de quimotripsina das sementes de erythrina velutina (EvCI)

Studies indicate that several components were isolated from medicinal plants, which have antibacterial, antifungal, antitumor and anti-inflammatory properties. Sepsis is characterized by a systemic inflammation which leads to the production of inflammatory mediators exacerbated by excessive activation of inflammatory cells and disseminated intravascular coagulation (DIC), in which the human neutrophil elastase plays an important role in its pathogenesis. Several epidemiological studies suggest that components of plants, especially legumes, can play a beneficial role in reducing the incidence of different cancers. A chymotrypsin inhibitor of Kunitz (Varela, 2010) was purified from seeds of Erythrina velutina (Mulungu) by fractionation with ammonium sulfate, affinity chromatography on Trypsin-Sepharose, Chymotrypsin-Sepharose and ion exchange chromatography on Resource Q 1 ml (GE Healthcare) in system FPLC / AKTA. The inhibitor, called EvCI, had a molecular mass of 17 kDa determined by SDS-PAGE. The purified protein was able to inhibit human neutrophil elastase (HNE), with an IC50 of 3.12 nM. The EvCI was able to inhibit both pathways of HNE release stimulated by PAF and fMLP (75.6% and 65% respectively). The inhibitor also inhibited leukocyte migration in septic mice about 87% and prolonged the time of coagulation and inhibition factor Xa. EvCI showed neither hemolytic activity nor cytotoxicity. EvCI showed a selective antiproliferative effect to HepG2 cell lines with IC50 of 0.5 micrograms per milliliter. These results suggest EvCI as a molecule antagonist of PAF / fMLP and a potential use in fighting inflammation related disorders, disseminated intravascular coagulation (DIC) and cancer

Clonagem e expressão do gene que codifica o inibidor de quimotripsina de Erythrina velutina WILLD. - caracterização e avaliação de seu potencial farmacológico

Proteinases are enzymes distributed widely founded in several organisms and perform many different functions, from maintaining homeostasis to the worsening of some diseases such as cancer, autoimmune diseases and infections. The proteins responsible of controlling the action of these enzymes are the inhibitors, that are classified based on their target proteases and are founded since simple organisms, such as bacteria, to higher organisms, such as larger plants and mammals. Plant proteinase inhibitors act by reducing or inactivating the activity of target proteases, thus, these proteins have been studied as potential tools in the treatment of diseases related to protease activities. In this context, an inhibitor of chymotrypsin from Erythrina velutina, called EvCI was previously purified and it was observed that this protein plays in vitro anticoagulant activity and anti-inflammatory activity in in vivo model. Aiming to reduce the environmental impact caused by the purification EvCI in high amounts and to facilitate the process of obtaining this protein, the recombinant chymotrypsin inhibitor from Eryhrina velutina was produced after cloning and expression in Escherichia coli. The bacteria were grown in LB medium and after induction of the expression this material was subjected to procedures for cell lysis and the product was applied on Nickel-affinity column. The proteins adsorbed were digested by thrombin and applied on Chymotrypsin-Sepharose affinity column, obtaining the purified inhibitor, named recEvCI. After electrophoresis, the recombinant inhibitor showed an approximately molecular mass of 17 kDa, and reduced the chymotrypsin and elastase activities in vitro. The recombinant inhibitor was sequenced and was found similar amino acids residues when compared to other inhibitors deposited in the database, with some modifications. recEvCI showed high stability under pH variations and reducing conditions, maintaining its activity around 80%. This protein increased the blood coagulation time in vitro by acting on the intrinsic pathway and did not show cytotoxicity against strains of mouse 3T3 fibroblasts and RAW 264.7 macrophages. recEvCI showed microbicide activity related to release of nitric oxide and consequently the activation of macrophages, futhermore having proinflammatory effects assessed by increased release of TNF-α. These results indicate that recEvCI can be biotechnologically used as a new tool in the control of coagulation-related diseases as well as can be an activating agent of the immune system in immunosuppressed individuals

Um novo inibidor de tripsina multifuncional de sementes de Erythrina velutina: caracterizacao e propriedades farmacologicas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Crescimento inicial e morfologia foliar em plantas de Enterolobium contortisiliquum (Vell.) Morong. E Erythrina velutina Mart. ex Benth, sob estresse hídrico

The Caatinga is the predominant vegetation type in semi-arid region of Brazil, where many inhabitants depend on hunting and gathering for survival, obtaining resources for: food and feed, folk medicine, timber production, etc. It‟s the dry ecosystem with highest population density in the world. The early stages of development are the most critical during the life cycle of a flowering plant and they‟re primordial to its establishment in environments exposed to water stress. Information about adjustments to the growth of the species, correlated with their studies of distribution in Seridó oriental potiguar, are an important ecological and economic standpoint, because they provide subsidies for the development of cultivation techniques, to programs of sustainable use and recovery of degraded areas. This thesis aimed to study the initial growth and foliar morphology in plants like Enterolobium contortisiliquum (Vell.) Morong. (tamboril) and Erythrina velutina Mart. ex Benth (mulungu), species of occurrence in the Caatinga, under water stress. After sowing and emergency, the seedlings were exposed to three water regimes: 450 (control), 225 (moderate stress) and 112.5 (severe stress) mm of water slide for 40 days. Seeding occurred in bags of 5 kg and after the establishment of seedlings thinning was carried out leaving a plantlet per bag. At the beginning the waterings occurred daily with distilled water, passing to be on alternate days after thinning. Twenty and forty days after the thinning seedlings collections were held to be done analysis of growth and biomass partition. When compared to the control group, the treatments with water stress showed reduction in the growth of the aerial part, growth of the greater root, number of leaves and leaflets, dry leaf area and total phytomass in both species, but in general, this effect was most marked for E. velutina. Regarding the partition of biomass, there were few changes throughout the experiment. Morphological changes in the leaves as a function of stress were not significant, however, there was a trend, in both species, to produce narrower leaves, that facilitate heat loss to the environment. It has not been possible to establish a positive relationship between inhibition of growth and distribution of species, whereas E. velutina is a species of most common occurrence in Seridó oriental potiguar. In this way, other aspects should be taken into account when studying the adaptation of species the dry environments, such as salinity, presence of heavy metals, wind speed, etc