Composição química, atividades inibidora da acetilcolinesterase e antifúngica de Pera glabrata (Schott) Baill. (Euphorbiaceae)

Pera glabrata (Schott) Baill. was selected for this study after showing a preliminary positive result in a screening of Atlantic Forest plant species in the search for acetylcholinesterase inhibitors and antifungal compounds. The bioassays were conducted with crude ethanol extract of the leaves using direct bioautography method for acetylcholinesterase and antifungal activities. This extract was partitioned with hexane, chloroform and ethyl acetate solvents. The active chloroform fraction was submitted to silica gel chromatography column affording 12 groups. Caffeine, an alkaloid, which showed detection limits of 0.1 and 1.0 µg for anticholinesterasic and antifungal activities, respectively, was isolated from group nine. After microplate analyses, only groups four, nine, 10, 11 and 12 showed acetylcholinesterase inhibitory activity of 40% or higher. The group 12 was purified by preparative layer chromatography affording four sub-fractions. Two sub-fractions from this group were analyzed by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. The first sub-fraction showed anticholinesterasic activity and contained two major compounds: 9-hydroxy-4-megastigmen-3-one (84%) and caffeine (6%). The second sub-fraction presented five major compounds identified as 9-hydroxy-4-megastigmen-3-one, isololiolide, (-) loliolide, palmitic acid and lupeol and did not show activity.Pera glabrata (Schott) Baill. foi selecionada para este estudo a partir de uma triagem de espécies vegetais da Mata Atlântica na busca de substâncias com atividades anticolinesterásica e antifúngica. A técnica da bioautografia direta foi utilizada para a detecção das atividades anticolinesterásica e antifúngica. O extrato etanólico bruto obtido das folhas foi particionado com hexano, clorofórmio e acetato de etila. A fração clorofórmica ativa foi fracionada por cromatografia em coluna de sílica gel fornecendo 12 grupos. Do grupo nove foi isolado o alcalóide cafeína com limites de detecção de 0,1 e 1,0 µg para as atividades anticolinesterásica e antifúngica, respectivamente. Após bioensaio em microplaca, somente os grupos quatro, nove, 10, 11 e 12 apresentaram inibição da acetilcolinesterase maior ou igual a 40%. O grupo 12 foi purificado por cromatografia em camada delgada preparativa de sílica gel fornecendo quatro sub-frações. Duas sub-frações deste grupo foram analisadas por cromatografia a gás-espectrometria de massas e cromatografia a gás com detector de ionização de chama. A primeira sub-fração contém dois compostos majoritários: 9-hidroxi-4-megastigmen-3-ona (78%) e cafeína (6%), e apresentou atividade anticolinesterásica. A segunda sub-fração contém cinco compostos principais identificados como 9-hidroxi-4-megastigmen-3-ona, isololiolida, (-) loliolida, ácido palmítico e lupeol e não apresentou atividade.FapespCNPq - PibicCapes - Prodo

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Alkaloids from Stems of Esenbeckia leiocarpa Engl. (Rutaceae) as Potential Treatment for Alzheimer Disease

Esenbeckia leiocarpa Engl. (Rutaceae), popularly known as guarantã, goiabeira, is a native tree from Brazil. Bioactivity-guided fractionation of the ethanol stems extract afforded the isolation of six alkaloids: leiokinine A, leptomerine, kokusaginine, skimmianine, maculine and flindersiamine. All isolated compounds were tested for acetyl cholinesterase inhibition, in vitro and displayed anticholinesterasic activity. The alkaloid leptomerine showed the highest activity (IC50 = 2.5 mM), similar to that of the reference compound galanthamine (IC50 = 1.7 mM). The results showed for the first time the presence of alkaloids leptomerine and skimmianine in E. leiocarpa (Engl.) with potent anticholinesterasic activity

Action of alkaloids of Acosmium subelegans (Mohlenbr.) Yakovlev in Ca2+ influx and mobilization in primary cultures of skeletal and cardiac muscles

A Acosmium subelegans (MohlenbrJ Yakovlev, conhecida popularmente como perobinha do campo, e utilizada na medicina popular principalmente como antiespasmodica, sedativa febrifuga na epilepsia, nos acessos da asma e na~ coqueluche' Esta especie foi considerada prioritaria no 1° Projeto de Plantas Medicinais (CEME-MS) na busca de medicamentos autoctones a partir de produtos naturais de uso popular. Estudos anteriores de nosso laboratorio mostraram que o extrato aquoso (EA) e a fracao alcaloidica (FALK) da A. subelegans potenciam a contracao dos musculo diafragma estimulado diretamente na presenca de d-tubocurarina. Na dependencia da concentracao do extrato ou da fracao, e do tempo de incubacao, a i potenciacao foi seguida de inibicao das contracoes. As fracoes semi-purificadas da FALK, F-1, F-3 e F-5 mantiveram a atividade potenciadora das contracoes no musculo esqueletico (Cardoso 1998). Estes resultados sugeriram uma provavel interferencia do EA e fracoes da A. subelegans na homeostasia intracelular de calcio. Esta tese teve por objetivo aprofundar os estudos dos mecanismos de acao das fracoes semi-purificadas e das substancias isoladas da A. subelegans no influxo e na mobilizacao de calcio em culturas de musculos esqueletico e cardiaco. Os resultados obtidos sao apresentados a seguir: 1. A FALK da A. subelegans apresentou atividade inotropica positiva e/ou negativa em atrio esquerdo de rato estimulado eletricamente. Concentracoes menres de FALK(30 e 100 g/mL) aumentaram a concentracao atrial basal. Este efeito inotropico positivo nao foi bloqueado por propanolol. Ja, concentracoes maiores de FALK (300 e 600 g/mL) diminuiram as concentracoes e induziram falhas na concentracao atrial. Estes resultados descartaram a participacao dos receptores -adrenergicos no efeito inotropico positivo da fracao e indicaram uma possivel acao intracelular dos alcaloides. 2. A FALK(30 a 300 g/mL) prolongou de forma concentracao- e tempo- dependente o periodo refrataria efetivo atrial (PRE). 3. A FALK foi purificada em CLEAR, originando 5 fracoes semi-purificadas identificadas de F-1 a F-5a(au)BV UNIFESP: Teses e dissertaçõe

Preclinical evaluation of Luffa operculata Cogn. and its main active principle in the treatment of bacterial rhinosinusitis

Abstract Introduction The prevalence of rhinosinusitis is quite high. Despite the widespread use of antibiotics for rhinosinusitis, there are other forms of treatment, including phytotherapy. One of the most widely used herbal medicines for treatment of rhinosinusitis is Luffa operculata. Objective This study aimed to evaluate the efficacy of topical nasal solution of the aqueous extract of L. operculata, determining the toxicity to its use and identifying the active principles presented in the aqueous extract. The secondary objective was to evaluate the action of active principles on bacteria commonly involved in acute rhino sinusitis. Methods The study was conducted in experimental model of sinusitis. Three different concentrations of L. operculata were used as local treatment of rhino sinusitis. The results were compared with those observed in control groups that received nasal saline solution. Histological examination of the liver, kidney, spleen, myocardium, brain and lungs of all animals evaluated the toxicity of L. operculata. The aqueous extract used was subjected to chromatographic analysis and an active principle was isolated and tested for in vitro inhibition of bacterial colonies usually found in rhino sinusitis. Results Intranasal treatment of sinusitis with L. operculata showed better clinical evolution than control group. Statistically significant difference (p > 0.10) between the treated group and the control group was observed in the histologic evaluation for inflammatory pattern. The aqueous extract of L. operculata used presented a predominance of 2,3-dicafeoilglicaric acid, a substance not yet described in the literature. There was a significant difference in bacterial growth of Streptococcus pyogenes on blood-agar plates when under the influence of both the aqueous extract and the active substance. Conclusion Topical nasal solution of the aqueous extract of L. operculata is effective compared to the application of saline solution for the treatment of bacterial rhinosinusitis in an experimental model. L. operculata determined in vitro inhibition of growth of S. pyogenes

Preclinical evaluation of Luffa operculata Cogn. and its main active principle in the treatment of bacterial rhinosinusitis

<div><p>Abstract Introduction The prevalence of rhinosinusitis is quite high. Despite the widespread use of antibiotics for rhinosinusitis, there are other forms of treatment, including phytotherapy. One of the most widely used herbal medicines for treatment of rhinosinusitis is Luffa operculata. Objective This study aimed to evaluate the efficacy of topical nasal solution of the aqueous extract of L. operculata, determining the toxicity to its use and identifying the active principles presented in the aqueous extract. The secondary objective was to evaluate the action of active principles on bacteria commonly involved in acute rhino sinusitis. Methods The study was conducted in experimental model of sinusitis. Three different concentrations of L. operculata were used as local treatment of rhino sinusitis. The results were compared with those observed in control groups that received nasal saline solution. Histological examination of the liver, kidney, spleen, myocardium, brain and lungs of all animals evaluated the toxicity of L. operculata. The aqueous extract used was subjected to chromatographic analysis and an active principle was isolated and tested for in vitro inhibition of bacterial colonies usually found in rhino sinusitis. Results Intranasal treatment of sinusitis with L. operculata showed better clinical evolution than control group. Statistically significant difference (p > 0.10) between the treated group and the control group was observed in the histologic evaluation for inflammatory pattern. The aqueous extract of L. operculata used presented a predominance of 2,3-dicafeoilglicaric acid, a substance not yet described in the literature. There was a significant difference in bacterial growth of Streptococcus pyogenes on blood-agar plates when under the influence of both the aqueous extract and the active substance. Conclusion Topical nasal solution of the aqueous extract of L. operculata is effective compared to the application of saline solution for the treatment of bacterial rhinosinusitis in an experimental model. L. operculata determined in vitro inhibition of growth of S. pyogenes.</p></div

Alkaloids from Stems of Esenbeckia leiocarpa Engl. (Rutaceae) as Potential Treatment for Alzheimer Disease

Esenbeckia leiocarpa Engl. (Rutaceae), popularly known as guaranta, goiabeira, is a native tree from Brazil. Bioactivity-guided fractionation of the ethanol stems extract afforded the isolation of six alkaloids: leiokinine A, leptomerine, kokusaginine, skimmianine, maculine and flindersiamine. All isolated compounds were tested for acetyl cholinesterase inhibition, in vitro and displayed anticholinesterasic activity. The alkaloid leptomerine showed the highest activity (IC(50) = 2.5 mu M), similar to that of the reference compound galanthamine (IC(50) = 1.7 mu M). The results showed for the first time the presence of alkaloids leptomerine and skimmianine in E. leiocarpa (Engl.) with potent anticholinesterasic activity.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[03/02176-7]CNPqPRODOC/CAPES of Instituto de Botanica of Sao Paulo, Brazi

Screening for antifungal, DNA-damaging and anticholinesterasic activities of Brazilian plants from the Atlantic Rainforest: Ilha do Cardoso State Park

Extratos brutos de 17 espécies de plantas coletadas em região de Mata Atlântica no Estado de São Paulo (Brasil) foram avaliadas para as atividades: antifúngica, no reparo do DNA e inibição da acetilcolinesterase. Dos 34 extratos obtidos de folhas e galhos das plantas analisadas para a atividade antifúngica com Cladosporium sphaerospermum e C. cladosporioides, 26,5% foram ativos. Todavia, apenas o extrato das folhas de Cabralea canjerana inibiu fortemente o crescimento dos dois fungos. No ensaio de reparo do DNA com linhagens mutantes de Saccharomyces cerevisiae, 11,7% dos extratos foram ativos, sendo que, 100% destes foram seletivos para o mecanismo de reparo do DNA envolvendo topoisomerase II. Das 17 espécies analisadas, 12 demonstraram atividade anticolinesterásica no ensaio qualitativo sobre cromatografia de camada delgada (CCD). No entanto, apenas os extratos de Tetrastylidium grandifolium (galhos) e Sloanea guianensis (folhas e galhos) apresentaram atividade anticolinesterásica maior que 50% no ensaio quantitativo.Crude extracts from 17 plant species collected from an Atlantic Forest region in the State of São Paulo (Brazil) have been screened for antifungal, DNA-damaging and acetylcholinesterase inhibiting activities. of the 34 extracts obtained from leaves and stems of plants assayed for antifungal activity with Cladosporium sphaerospermum and C. cladosporioides 26.5% were active. However, only the extract of leaves of Cabralea canjerana showed a strong inhibition of both fungi. The DNA-damaging assay with mutant strains of Saccharomyces cerevisiae resulted in 11.7 % of the extracts being active whereas 100% of them showed selectivity for the DNA-repair mechanism of topoisomerase II. of the 17 species analysed, 12 showed anticholinesterasic activity in TLC assay. However, only extracts from Tetrastylidium grandifolium (stems) and Sloanea guianensis (leaves and stems) inhibited acetylcholinesterase activity more than 50% in quantitative assay.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES