Inhibitory effects of compounds isolated from the dried branches and leaves of murta (Myrceugenia euosma) on lipid accumulation in 3T3-L1 cells.

As obesity is a global health concern the demand for anti-obesity drugs is high. In this study, we investigated the anti-obesity effect of the dried branches and leaves of murta (Myrceugenia euosma Legrand, Myrtaceae). A methanol extract of the dried branches and leaves of murta inhibited adipogenesis in 3T3-L1 cells. Three known flavanones-cryptostrobin (1), pinocembrin (4), and 5,7-dihydroxy-6,8-dimethylflavanone (6), and three chalcones-2',6'-dihydroxy-3'-methyl-4'-methoxychalcone (2), pinostrobin chalcone (3), and 2',6'-dihydroxy-4'-methoxy-3',5'-dimethylchalcone (5) were isolated from the active fraction. Structures of these compounds were identified using various spectral data. Each of these compounds also inhibited adipogenesis in 3T3-L1 cells. In particular, compound 3 was a more potent inhibitor of triglyceride accumulation than the positive control berberine. Gene expression studies revealed that treatment of 3T3-L1 cells with 3 lowers the expression levels of CCAAT/enhancer-binding protein α and peroxisome proliferator activator γ2 during adipogenesis without affecting cell viability. Treatment of 3T3-L1 cells with 3 reduced the expression levels of mRNAs encoding sterol regulatory element-binding protein 1c and several lipogenic enzymes, including fatty acid synthase and stearoyl CoA desaturase-1. These results indicate that the methanol extract and compounds isolated from the dried branches and leaves of murta exert their anti-obesity effects through the inhibition of adipogenesis.

Green Propolis: Thirteen Constituents of Polar Extract and Total Flavonoids Evaluated During Six Years through RP-HPLC.

In order to verify the chemical qualities of polar extract of Green Propolis produced in the State of Minas Gerais, Brazil, was analyzed by means of RP-HPLC, the concentration of eleven flavonoids, Artepillin C and p-coumaric acid were analyzed by means of RP-HPLC. Samples were collected in the months of February, March, May, July, September, and October, during a period of six years (2008 to 2013) and the results compared with purchased standards. Artepillin C was the main constituent (80-90%) found in all samples. Lower concentrations of p-coumaric acid, chrysin and kaempferide were observed in March, and of ricin and galangin in September. Respectively, these months correspond to the end of Summer and Spring period in Brazil. The variation in concentration of the analyzed constituents always occurred in the same month, during the entire six years of evaluation period. All thirteen constituents of Green Propolis were detected throughout the study period, and the average concentration of each one was similar in relation to the respective monthly collection period of each year. Due to similarities among chemical constituents of Green Propolis with those present in B. dracuncufolia, this plant was identified as being the principal source of Green Propolis.

Green propolis modulates gut microbiota, reduces endotoxemia and expression of TLR4 pathway in mice fed a high-fat diet.

Due to the various beneficial effects attributed to propolis, which include anti-inflammatory and anti-bacterial infection properties, the objective of the study was to evaluate the effect of propolis supplementation on the composition of the intestinal microbiota and its anti-inflammatory action. Forty male C57BL/6 mice were fed either a standard diet (control), a high-fat (HF) diet, or a high-fat diet supplemented with 0.2% crude propolis (HFP) for 2 or 5weeks prior to sacrifice. Blood samples were collected for the determination of lipopolysaccharide (LPS) and classical biochemical parameters. Expression of the TLR4 pathway in muscle, and DNA sequencing for the 16S rRNA of the gut microbiota were performed. The HF diet increased the proportion of the phylum Firmicutes and inflammatory biomarkers, while supplementation with propolis for five weeks rendered the microbiota profile nearly normal. Consistently with the above, the supplementation reduced levels of circulating LPS and down-regulated the TLR4 pathway and inflammatory cytokine expressions in muscle. Moreover, propolis improved such biochemical parameters as serum triacylglycerols and glucose levels. The data suggest that propolis supplementation reduces inflammatory response and endotoxemia by preventing dysbiosis in mice challenged with a high-fat diet.

Effect of brazilian propolis on exacerbation of respiratory syncytial virus infection in mice exposed to tetrabromobisphenol a, a brominated flame retardant.

Tetrabromobisphenol A (TBBPA), a brominated flame retardant, has been found to exacerbate pneumonia in respiratory syncytial virus- (RSV-) infected mice. We examined the effect of Brazilian propolis (AF-08) on the exacerbation of RSV infection by TBBPA exposure in mice. Mice were fed a powdered diet mixed with 1% TBBPA alone, 0.02% AF-08 alone, or 1% TBBPA and 0.02% AF-08 for four weeks and then intranasally infected with RSV. TBBPA exposure increased the pulmonary virus titer and level of IFN- γ , a representative marker of pneumonia due to RSV infection, in the lungs of infected mice without toxicity. AF-08 was significantly effective in reducing the virus titers and IFN- γ level increased by TBBPA exposure. Also, AF-08 significantly reduced proinflammatory cytokine (TNF- α and IL-6) levels in the lungs of RSV-infected mice with TBBPA exposure, but Th2 cytokine (IL-4 and IL-10) levels were not evidently increased. Neither TBBPA exposure nor AF-08 treatment affected the anti-RSV antibody production in RSV-infected mice. In flow cytometry analysis, AF-08 seemed to be effective in reducing the ratio of pulmonary CD8a(+) cells in RSV-infected mice with TBBPA exposure. TBBPA and AF-08 did not exhibit anti-RSV activity in vitro. Thus, AF-08 probably ameliorated pneumonia exacerbated by TBBPA exposure in RSV-infected mice by limiting excess cellular immune responses.

Recent progress of propolis for its biological and chemical compositions and its botanical origin.

Propolis is the generic name given to the product obtained from resinous substances, which is gummy and balsamic and which is collected by bees from flowers, buds, and exudates of plants. It is a popular folk medicine possessing a broad spectrum of biological activities. These biological properties are related to its chemical composition and more specifically to the phenolic compounds that vary in their structure and concentration depending on the region of production, availability of sources to collect plant resins, genetic variability of the queen bee, the technique used for production, and the season in which propolis is produced. Many scientific articles are published every year in different international journal, and several groups of researchers have focused their attention on the chemical compounds and biological activity of propolis. This paper presents a review on the publications on propolis and patents of applications and biological constituents of propolis.

Efficacy of Brazilian Propolis against Herpes Simplex Virus Type 1 Infection in Mice and Their Modes of Antiherpetic Efficacies.

Ethanol extracts (AF-06, 07, and 08, 10 mg/kg) of Brazilian propolis were administered orally to cutaneously herpes simplex virus type 1 (HSV-1)-infected mice three times daily on days 0 to 6 after infection to evaluate their efficacies against HSV-1 infection and significantly limited development of herpetic skin lesions. AF-07 and 08 significantly reduced virus titers in brain and/or skin on day 4 without toxicity, but AF-08 had no anti-HSV-1 activity in vitro. AF-06 and 08 significantly enhanced delayed-type hypersensitivity (DTH) to inactivated HSV-1 antigen in infected mice. Oral AF-08-administration significantly augmented interferon (IFN)-γ production by HSV-1 antigen from splenocytes of HSV-1-infected mice, while direct exposure of splenocytes of infected mice to AF-06 significantly elevated IFN-γ production in vitro. Thus, AF-08 might have components that are active in vivo even after oral administration and those of AF-06 might be active only in vitro. Because DTH is a major host defense for intradermal HSV-1 infection, augmentation of DTH response by AF-06 or 08, directly or indirectly, respectively, may contribute to their efficacies against HSV-1 infection. In addition, AF-06 and 07 possibly contain anti-HSV-1 components contributing to their efficacies. Such biological activities of Brazilian propolis may be useful to analyze its pharmacological actions.

Anti-influenza virus activity of propolis in vitro and its efficacy against influenza infection in mice.

BACKGROUND: Propolis has been used worldwide as a dietary supplement to maintain and improve human health. We examined whether ethanol extracts of Brazilian propolis exhibit antiviral activity against influenza virus in vitro and in vivo. METHODS: Among 13 ethanol extracts screened in a plaque reduction assay, four showed anti-influenza virus activity. The anti-influenza efficacy of the four extracts was further examined in a murine influenza virus infection model. The mice were infected intranasally with influenza virus, and the four extracts were orally administered at 10 mg/kg three times daily for seven successive days after infection. RESULTS: In this infection model, only one extract, AF-08, was significantly effective at 10 mg/kg in reducing the body weight loss of infected mice. The doses of 2 and 10 mg/kg were also effective in prolonging the survival times of infected mice significantly, but 0.4 mg/kg was not. The anti-influenza efficacy of AF-08 at 10 mg/kg was confirmed in a dose-dependent manner in mice. AF-08 at 10 mg/kg significantly reduced virus yields in the bronchoalveolar lavage fluids of lungs in infected mice as compared with the control. The reduction of virus yields by AF-08 at 10 mg/kg significantly corresponded to those induced by oseltamivir at 1 mg/kg twice daily from day 1 to day 4 after infection. CONCLUSION: The Brazilian propolis AF-08 was indicated to possess anti-influenza virus activity and to ameliorate influenza symptoms in mice. AF-08 may be a possible candidate for an anti-influenza dietary supplement for humans.

Production of xylanase and CMCase on solid state fermentation in different residues by Thermoascus aurantiacus miehe

O emprego de residuos como matéria prima é importante como estrategia governamental e para o balanco ambiental. O propósito deste trabalho foi estudar a producão de CMCase e xilanase de uma linhagem de Thermoascus aurantiacus isolado de solo brasileiro em fermentacão em estado sólido (SSF) usando diferentes resíduos agrícolas (farelo de trigo, bagaco de cana, bagaco de laranja, sabugo de milho, grama verde, grama seca, serragem de eucalipto e palha de milho) como substratos sem enriquecimento dos meios e caracterizar as enzimas. O estudo das enzimas hemiceluloliticas extracelulares mostrou que o fungo T. arantiacus é mais xilanolítico do que celulolítico. Ele produziu maiores níveis das enzimas em meios contendo sabugo de milho, grama e palha de milho. Todas as enzimas foram estáveis por 24 h à temperatura ambiente numa ampla faixa de pH (3,0 ù 9,0) e também foram estáveis a 60ºC por 1 h. O pH ótimo e temperatura ótima para xilanase e CMCase foram 5,0ù 5,5 e 5,0 e 75ºC, respectivamente. O microrganismo cresceu muito bem estacionariamente no meio simples, de baixo custo. As enzimas estáveis secretadas extracelularmente apresentam as características necessárias para sua aplicacão industrial.

Composição química de Baccharis dracunculifolia, fonte botânica das própolis dos estados de São Paulo e Minas Gerais / Chemical composition of Baccharis dracunculifolia, the botanical source of propolis from the states of São Paulo and Minas Gerais, Brazil

A própolis é uma substância resinosa coletada pelas abelhas de diversas partes das plantas. Sua composição depende da época, vegetação e local de coleta. Análises fitoquímicas de própolis produzidas de Baccharis dracunculioflia (alecrim-do-campo) por abelhas Apis mellifera africanizada nos estados de São Paulo e Minas Gerais foram feitas por espectrofotometria na região U.V.-Visível, CCDAE-FR, CLAE-FR e CG-EM. Uma característica dos compostos fenólicos das própolis analisadas e da espécie vegetal de B. dracunculiofolia foi a alta proporção de artepilin C e outros derivados do ácido cinâmico. Com base nas evidências fitoquímicas, B. dracunculifolia foi identificada como a principal fonte vegetal das própolis produzidas nos estados de São Paulo e Minas Gerais.

Conversão de daidzina e genistina de soja por b-glicosidase de Aspergillus oryzae / Conversion of soy daidzein and genistein by beta-glucosidase of Aspergillus oryzae

Avaliou-se a capacidade de conversão de isoflavonas glicosiladas às suas formas agliconas pelo uso de B-glicosidase de Aspergillus oryzae ATCC 22786. A produção de daidzeína e genisteína foi acompanhada por 96 h de fermentação em estado sólido, usando-se farelo de soja e suspensão de esporos do fungo a 30ºC. Notou-se a conversão de glicosil-isoflavonas após 24 h e redução significativa na suas quantidade depois de 48 h de fermentação. A atividade enzimática acompanhou a formação de produto até 72 h (0,2 Ul/mL), apresentando leve diminuição em 96 h (0,19Ul/mL). Após 48 h obteve-se conversão total de daidzina e genistina, com formação de 551,1 e 127,2 ug/g de daidzeína e genisteína, respectivamente. A produção de genisteína, no entanto, mostrou-se significativamente superior em 24 h de fermentação (289,3 ug/g). Concluiu-se que a B-glicosidase produzida por Aspergillus oryzae ATCC 22786 foi capaz de converter glicosilsólido a 30ºC.

Própolis produzida no sul do Brasil, Argentina e Uruguai: evidências fitoquímicas de sua origem vegetal / Propolis produced in south Brazil, Argentine and Uruguay: phytochemical evidence for the plant origin

A própolis é uma substância resinosa coletada pelas abelhas de diversas partes da planta. Sua composiçäo depende da época, vegetaçäo e área de coleta. Própolis produzida por abelhas Apis mellifera africanizada do sul do Brasil, Argentina e Uruguai foram investigadas por meio de técnicas cromatográficas. De acordo com os resultados obtidos por CCDAE-FR, CLAE-FR e CG-EM, as própolis do sul do Brasil foram classificadas em 5 grupos distintos. Com base nas evidências fitoquímicas, Populus alba foi identificada como a principal fonte da própolis do grupo 3 do sul do Brasil, Argentina e Uruguai

Biochemical characterization of a microbial glucosyltransferase that converts sucrose to palatinose

Foi isolada de fruta deteriorada uma linhagem de klebsiella sp que transforma sacarose em palatinose. Foi verificado que a linhagem produz uma glicosiltransferase intracelular que transforma sacarose para palatinose. A enzima de Klebsiella sp foi purificada por fracionamento com sulfato de amônio e cromatografia em colunas de DEAE-Sephadex A-50 e CM-celulose. A enzima purificada apresentou ótima atividade a 35ºC e na faixa de pH 6,0 a 6,5. A enzima foi inibida por Hg2+ e Ag+, mas näo foi inibida por p-cloromercuribenzoato. Os valores de km e Vmax da enzima purificada foram respectivamente 120mM e 110µg palatinose formada por minuto por ml de enzima para o substrato sacarose. O peso molecular da enzima foi estimado em 74.000 Da. A enzima converteu 4 por cento (p/w) de sacarose para isomaltulose com uma eficiência de 86 por cento a 25ºC, pH 6,5

Screening of Beta-fructofuranosidase producing microorganisms por production of fructooligosaccharides and studies of some enzyme properties

Uma linhagem de levedura, produtora de Beta-frutofuranosidade de alta atividade, foi isolada em folha tropical denominada Araticum (Anona densicoma) tendo sido identificada como Aureobasidium pellulans. Esta linhagem produziu Beta-frutofruanosidade extra e intracelularmente que catalizam a produçäo de frutooligossacarídeos a partir da sacarose. O pH e temperatura ótimas de atividade de enzima foram 5.2 e 55ºC, respectivamente. Sua termoestabilidade foi de 55ºC. Esta enzima converteu sacarose a 61.2 por cento de frutooligossacarídeos quando se utilizou sacarose como substrato na concentraçäo de 50 por cento

Produçäo de aroma de frutas por espécies de Neurospora isoladas de beiju / Production of fruity aroma by Neurospora species isolated from beiju

Oito linhagens de Neurospora sp. foram isoladas de beiju, em várias regiöes do Estado do Maranhäo, Brasil. As linhagens apresentaram um agradável aroma de frutas no meio da cultura, enquanto que, as linhagens de Neurospora da coleçäo de cultura NRRL e outras linhagens de Neurospora isoladas de solo na regiäo de Säo Paulo, näo produziram aroma de frutas. Foi detectado por Cromatografia Gasosa por "Headspace" fque o composto responsável pelo aroma de frutas era o etil hexanoato. Além disto, as linhagens de Neurospora sp. isoladas do Maranhäo produziram 3-metil-1-butanol, 1-octen-3-ol, acetato de etila e etanol.

Produçäo de ciclodextrina glicosiltransferase por Bacillus lentus alcalofílico / Cyclodextrin glycosyltransferase production by alkalophilic bacillus lentus

Resumo: Uma linhagem de Bacillus lentus mesofílico e alcalofílico contendo alta atividade extracelular de ciclodetrina glicosiltransferese(CGtase, E.C.2.41.19), foi obtida de uma coleçäo de microrganismos isolados de solos da cidade de Campinas(SP), utilizando-se um meio de cultivo alcalino.Uma preparaçäo bruta da CGTase foi obtida cultivando-se o microrganismo por 4 dias a 37ºC, sob aeraçäo em meio líquido contendo 2 por cento amido solúvel, 0, 5 por cento peptona, 0, 5 por cento extrato de levedura, 0, 1 por cento K2HPO4, 0, 02 por cento MgSO4.7H2O e 1 por cento Na2CO3(pH 10, 3).Os ótimos de pH e temperatura da enzima para a formaçäo de ciclodexinas, situaram-se entre 6, 5 a 7, 5 e 45 a 55ºC, respectivamente.As ciclodextrinas , ß e foram produzidas numa proporçäo de 1: 67: 1, 6, respectivamente.

Melhoramento de Bacillus amyloliquefaciens por DNA transformante e fusäo de protoplastos para produçäo de amilase e proteases / Improvement of Bacillus amyloliquefaciens by DNA transformation and protoplasts fusion for amylase and proteases production

Utilizando-se Bacillus amyloliquefaciens ATCC 2342 e o B. natto, isolado do produto comercial "natto", respectivamente, bons produtores de alfa-amilase e proteases, conseguiu-se aumentar os níveis de produçäo destas enzimas através das técnicas de DNA transformante e fusäo de protoplastos. O melhoramento genético por transformaçäo foi mais eficiente do que aquele por fusäo de protoplastos