Green tea extract rich in epigallocatechin-3-gallate prevents fatty liver by ampk activation via lkb1 in mice fed a high-fat diet

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Changes in the oxidative stress biomarkers in liver of streptozotocin-diabetic rats treated with combretum lanceolatum flowers extract

Aims: The study investigated the in vivo antioxidant activity and the in vitro radical scavenging capacity of the Combretum lanceolatum Pohl (Combretaceae) flowers ethanolic extract (ClEtOH) in streptozotocin-diabetic rats. Place and Duration of Study: Department of Chemistry, Federal University of Mato Grosso, Cuiabá, Brazil; between February 2012 and December 2012. Methodology: Male Wistar rats were divided into four groups: Normal rats treated with water/vehicle (N); diabetic rats treated with water (DC); diabetic rats treated with 250 mg/kg (DT250) or with 500mg/kg (DT500) of ClEtOH. After 21 days of treatment, liver samples were used for the analysis of the oxidative stress biomarkers and activity of antioxidant enzymes. In vitro radical scavenger capacity was investigated by the following methods: DPPH radical scavenging, ABTS radical cation decolorization and crocin bleaching assays. Results: Significant oxidative stress was observed in liver of DC, since the malondialdehyde (MDA, biomarker of lipoperoxidation) levels were increased in comparison with N. Increased activities of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also observed in DC, which could represent a compensatory mechanism against oxidative stress. Glutathione (GSH) levels were lower and similar between N and DC. The MDA levels were significantly decreased in liver of rats from DT250 and DT500, reaching levels similar those of N, suggesting that ClEtOH prevented lipoperoxidation. The treatment of diabetic rats with ClEtOH also increased the GSH levels, as well as increased the GSH-Px activity, and did not change the SOD activity. The results of in vitro radical scavenging capacity indicated that ClEtOH is highly active. Conclusion: These findings indicate that ClEtOH has antioxidant properties in liver of diabetic rats, decreasing lipoperoxidation and increasing the endogenous antioxidant responses. Both the antihyperglycemic effect and the capacity to scavenge free radicals may be related to the antioxidant activity of ClEtOH in diabetes.

Cinética micelial dos fungos comestíveis Pleurotus ostreatus e Lentinula edodes em resíduos lignocelulósicos

The objective of this work was to evaluate the mycelial growth of 2 edible fungi (Pleurotus ostreatus and Lentinula edodes) in 6 culture media [(malt-agar, sawdustdextrose-agar-marupá (SDA-MA), sawdust-dextrose-agar-cajuí (SDA-CA), sawdust-dextrose-agaraçaí (SDA-AÇA), sawdust-dextrose-agar-banana 50% (BAN 50%) and sawdust-dextrose-agar-banana 100% (BAN 100%)], in Petri dishes. The experimental design was totally randomized, in a 6x2 factorial scheme. Each treatment consisted of six repetitions in 1 Petri dish, totaling 72 experimental units. It was verified that P. ostreatus presented better mycelial development (81.00; 64.66; 81.00; 50.16 and 33.33mm for SDA-MA, SDA-CA, SDA-AÇA, BAN 50% and BAN 100%, respectively) than L. edodes (32.00; 31.66; 27.66; 37.33 and 21.83mm for SDA-MA, SDA-CA, SDA-AÇA, BAN 50% and BAN 100%, respectively). It was also verified that there was no advantage for L. edodes in relation to mycelial growth, when media based on residues were used, compared to malt-agar medium (control), which obtained the best performance (62.17mm). As for P. ostreatus, SDA-MA and SDA-AÇA medium presented the highest growth averages (81 mm), representing a growth increase of 34% in relation to the control medium (malt-agar), whose growth average was 60.33mm. Thus, the residues tested present potential to be used in fungiculture, especially for the cultivation of P. ostreatus.

Cultivo de Lentinula edodes e Pleurotus ostreatus em bagaço de cana-de-açúcar

Pós-graduação em Microbiologia - IBILCE

Potencial antibacteriano de cogumelos das espécies Lentinus edodes, Agaricus bisporus e Agaricus blazei com enfoque no Helicobacter pylori

A busca por novos agentes antimicrobianos é muito importante, uma vez que as bactérias possuem um alto potencial de recombinação genética, levando a um aumento de cepas resistentes aos fármacos antimicrobianos já existentes. Nesse contexto, os cogumelos podem ser uma alternativa, uma vez que extratos e constituintes químicos destes apresentam atividade frente a diversos isolados patogênicos e utilizados na medicina popular. Dentre as espécies de maior interesse estão o Agaricus blazei (cogumelo do sol), Agaricus bisporus (champignon de Paris) e Lentinus edodes (shitake). O Helicobacter pylori é responsável por doenças gastrointestinais de extrema importância para a saúde pública, como úlceras, gastrites até tumores malignos. O tratamento mais utilizado nos controles deste micro-organismo não é eficaz. Os cogumelos aparecem como alternativa para novas pesquisas neste ramo podendo vir a ser utilizados no desenvolvimento de novo fitoterápico ou mesmo como adjuvante deste tratamento. Existem diversos métodos para determinar a atividade antimicrobiana, entre eles a microdiluição. Esta técnica vem sendo muito utilizada principalmente devido à sua sensibilidade e pequena quantidade de reagentes e amostras, o que possibilita um maior número de réplicas, aumentando a confiabilidade dos resultados. Em continuidade ao estudo do nosso grupo de pesquisa este trabalho objetiva determinar o potencial anti-H. pylori de espécies de cogumelos comestíveis instigados pela possibilidade de encontrar uma alternativa para o tratamento desse micro-organismos dada a importância do mesmo na saúde pública. A atividade antibacteriana dos diferentes extratos de Lentinus edodes, Agaricus bisporus, obtidos por infusão, ultrassom e maceração e de extratos purificados de Agaricus blazei e da substância isolada β-glucana foi determinada pela técnica de microdiluição a fim de obter a concentração inibitória mínima (CIM) frente às...

Potencial antibacteriano de cogumelos das espécies Lentinus edodes, Agaricus bisporus e Agaricus blazei com enfoque no Helicobacter pylori

A busca por novos agentes antimicrobianos é muito importante, uma vez que as bactérias possuem um alto potencial de recombinação genética, levando a um aumento de cepas resistentes aos fármacos antimicrobianos já existentes. Nesse contexto, os cogumelos podem ser uma alternativa, uma vez que extratos e constituintes químicos destes apresentam atividade frente a diversos isolados patogênicos e utilizados na medicina popular. Dentre as espécies de maior interesse estão o Agaricus blazei (cogumelo do sol), Agaricus bisporus (champignon de Paris) e Lentinus edodes (shitake). O Helicobacter pylori é responsável por doenças gastrointestinais de extrema importância para a saúde pública, como úlceras, gastrites até tumores malignos. O tratamento mais utilizado nos controles deste micro-organismo não é eficaz. Os cogumelos aparecem como alternativa para novas pesquisas neste ramo podendo vir a ser utilizados no desenvolvimento de novo fitoterápico ou mesmo como adjuvante deste tratamento. Existem diversos métodos para determinar a atividade antimicrobiana, entre eles a microdiluição. Esta técnica vem sendo muito utilizada principalmente devido à sua sensibilidade e pequena quantidade de reagentes e amostras, o que possibilita um maior número de réplicas, aumentando a confiabilidade dos resultados. Em continuidade ao estudo do nosso grupo de pesquisa este trabalho objetiva determinar o potencial anti-H. pylori de espécies de cogumelos comestíveis instigados pela possibilidade de encontrar uma alternativa para o tratamento desse micro-organismos dada a importância do mesmo na saúde pública. A atividade antibacteriana dos diferentes extratos de Lentinus edodes, Agaricus bisporus, obtidos por infusão, ultrassom e maceração e de extratos purificados de Agaricus blazei e da substância isolada β-glucana foi determinada pela técnica de microdiluição a fim de obter a concentração inibitória mínima (CIM) frente às...

Pro-inflammatory effects of the aqueous extract of Echinometra lucunter sea urchin spines

The sea urchin, Echinometra lucunter, can be found along the Western Central Atlantic shores. In Brazil, it is responsible by circa 50% of the accidents caused by marine animals. The symptoms usually surpass trauma and may be pathologically varied and last differently, ranging from spontaneous healing in a few days, to painful consequences lasting for weeks. In this work, we have mimicked the sea urchin accident by administering an aqueous extract of the spine into mice and rats and evaluated the pathophysiological developments. Our data clearly indicate that the sea urchin accident is indeed a pro-inflammatory event, triggered by toxins present in the spine that can cause edema and alteration in the leukocyte-endothelial interaction. Moreover, the spine extract was shown to exhibit a hyperalgesic effect. The extract is rich in proteins, as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but also contains other molecules that can be analyzed by reversed phase high-performance liquid chromatography. Altogether, these effects corroborate that an E. lucunter encounter is an accident and not an incident, as frequently reported by the victims.

The effects of conjugated estrogen, raloxifene and soy extract on collagen in rat bones

Objective To evaluate the action of conjugated equine estrogen, raloxifene and isolated or combined genistein-rich soy extracts on collagen fibers in the bones of oophorectomized rats. Materials and methods Seventy female rats received testosterone propionate (0.1 mu g/g) on the 9th day after birth. At 6 months of age, the rats were administered the vehicle (propylene glycol, 0.5 ml/day), and ten of the rats were randomly chosen to comprise the non-oophorectomized control group (GI). The other 60 rats were ovariectomized and randomized into six groups of ten as follows: GII, vehicle; GIII, conjugated equine estrogen (CEE), 50 mu g/kg/day; GIV, raloxifene (RAL), 0.75 mg/kg/day; GV, genistein-rich soy extract (GSE), 300 mg/kg/day; GVI, CEE + GSE, 50 mu g/kg/day + 300 mg/kg/day; and GVII, CEE + RAL, 50 mu g/kg/day + 0.75 mg/kg/day. Three months after surgery, the drugs were administered for 60 consecutive days. All rats were euthanized, and their left tibiae were removed for histological routine. The histological sections were stained with hematoxylin-eosin, and picrosirius for evaluating bone microarchitecture. Types I and II collagen fibers were analyzed by immunofluorescence. Data analysis was carried out with ANOVA and Tukey's test. Results Collagen reduction was significant in the GIII animals when compared to the other groups (p < 0.05). There was no significant difference in the thickness of collagen fibers among the groups. There was a greater quantity of type III collagen in GVI than in the other groups. Conclusion Our data indicate that conjugated equine estrogen improves bone quality because it increases the quantity of type I collagen while reducing the quantity of thin collagen fibers. In addition, the combination of CEE and raloxifene or genistein-rich soy extract is not as efficient as CEE itself to improve bone quality.

Increased Antioxidant Content in Juice Enriched with Dried Extract of Pomegranate (Punica granatum) Peel

Antioxidants are compounds responsible for free radical scavenging in the body. They protect the organism from oxidative modification of cells and tissues. These modifications have been associated with degenerative diseases, atherosclerosis and carcinogenesis. Punica granatum displays high antioxidant potential due to the presence of phenolic compounds, which are capable of disease prevention. The present study showed the highest antioxidant activity in pomegranate peel than in seeds and pulp. Based on these results, pomegranate peel was used to produce dried extract that was added to commercial tomato juice and orange juice with strawberries. Analysis to determine the content of phenolic compounds and antioxidant activity was performed on pomegranate pulp, seeds and peel and in juices enriched with dried extract of pomegranate peel. The dried extract was responsible for a significant increase in antioxidant activity of the juices, proportional to the concentrations added. However, although both flavors of enriched juices displayed high antioxidant levels, the samples with higher dried extract concentrations received the lowest scores from sensory analysis participants due to the characteristic astringent flavor of pomegranate peels. Therefore, to obtain greater acceptance in the consumer market, we concluded that the maximum addition of dried pomegranate peel extract is 0.5% in tomato juice and orange juice with strawberries.

A new HPLC-DAD-MS/MS method for the simultaneous determination of major compounds in the crude extract of Lychnophora salicifolia Mart. (Brazilian arnicao) leaves: Application to chemical variability evaluation

Lychnophora salicifolia Mart., which occurs in the Brazilian Cerrado in the states of Bahia and Minas Gerais as well as in the southeast of the state of Goias, is the most widely distributed and also the most polymorphic species of the genus. This plant is popularly known to have anti-inflammatory and analgesic activities. In this work, we have studied the variation in terms of polar metabolites of ninety-three Lychnophora salicifolia Mart, specimens collected from different regions of the Brazilian Cerrado. Identification of the constituents of this mixture was carried out by analysis of the UV spectra and MS data after chromatographic separation. Twenty substances were identified, including chlorogenic acid derivatives, a flavonoid C-glucoside, and other sesquiterpenes. The analytical method was validated, and the reliability and credibility of the results was ensured for the purposes of this study. The concentration range required for analysis of content variability within the analyzed group of specimens was covered with appropriate values of limits of detection and quantitation, as well as satisfactory precision and recovery. A quantitative variability was observed among specimens collected from the same location, but on average they were similar from a chemical viewpoint. In relation to the study involving specimens from different locations, there were both qualitative and quantitative differences among plants collected from different regions of Brazil. Statistical analysis revealed that there is a correlation between geographical localization and polar metabolites profile for specimens collected from different locations. This is evidence that the pattern of metabolites concentration depends on the geographical distribution of the specimens. (C) 2012 Elsevier B.V. All rights reserved.

PHYSIOLOGICAL CHANGES IN SEEDS AND ANTIOXIDANT METABOLISM OF LETTUCE SEEDLING EXPOSED TO THE ACTION OF Zantedeschia aethiopica Spreng. LEAF EXTRACT

This work aimed to evaluate the influence of different concentrations of Zantedeschia aethiopica Spreng. extract on the physiological performance of the seed and on the response of the antioxidant metabolism of lettuce seedlings. The treatments consisted of leaves extracts from Z. aethiopica at concentrations of 0, 6, 12, 25 and 50%. Germination, first germination count, germination speed and index, length of shoot and radicle, seedling total dry mass, chlorophyll content, activity of superoxide dismutase, catalase and ascorbarte peroxidase enzymes, lipid peroxidation, hydrogen peroxide quantification and seedling emergence, length of organs, and total dry mass of seedlings were evaluated. The percentage of germination, the length of the shoot and radicle of seedlings and the total dry mass of seedlings grown in the greenhouse were reduced as the concentration of the extract increased. There were increases of electrical conductivity, of superoxide dismutase, catalase and ascorbate peroxidadase enzymes and the amount of hydrogen peroxide and lipid peroxidation in seedlings with increasing extract concentration. The extract reduced the physiological quality of lettuce seeds and induced an increased production of hydrogen peroxide in seedlings, which increased the activity of antioxidant enzymes that were not effective in tissue detoxification, resulting in cellular damage and increased numbers of abnormal seedlings.

Processing of Apeiba tibourbou Aubl. Extract Via Spray Drying

The effects of drying air inlet temperature (IT) and concentration of Aerosil 200 (C-A) on several properties of spray-dried Apeiba tibourbou extracts were investigated following a 3(2) full factorial design. Powder recovery varied from 9.83 to 46.95% and dried products showed moisture contents below 7%. Although the spray-dried products lost some of their polyphenols, they still present excellent antioxidant activity, opening perspectives for its use to medicinal purpose. C-A exerted a key role on the properties of spray-dried extracts, while IT did not present a significative influence. Aerosil (R) 200 proved to be an interesting alternative as an excipient for the drying of the herbal extract, even at intermediate concentrations such as 15%. The best combination of conditions to use for obtaining dry A. tibourbou extracts with adequate physicochemical and functional properties involves an IT of 100 degrees C and a C-A of 15%.

Cytotoxicity evaluation of Curcuma zedoaria (Christm.) Roscoe fluid extract used in oral hygiene products

Objective. This in vitro study evaluated the cytotoxic effects of the Curcuma zedoaria (Christm.) Roscoe (popular name: zedoary) fluid extract, as used in preparations for oral hygiene, mostly for anti-septic purposes. Materials and methods. The cell viability and cell growth were assessed by Trypan blue dye exclusion assay using the LMF cell line derived from oral mucosa. Cell viability (short-term assay) was measured 0, 6, 12 and 24 h after contact with the fluid extract. Cell growth (long-term assay) was analyzed in 1, 3, 5 and 7 days. The experimental groups were those testing the fluid extract obtained from the zedoary rhizome and the extractor liquid (ethanol 70 degrees GL) in the concentrations of 0.01-0.0001% v/v. Fresh DMEM were used in the control cultures. Results. Short-term assay-all studied cultures maintained stable cell viability; Long-term assay-there was progressive cell growth in all studied cultures. Conclusion. According to the results, the zedoary fluid extract presents low cytotoxicity and probably can be used in the oral hygiene products.

Biochemical Analysis of a Castor Bean Leaf Extract and its Insecticidal Effects Against Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)

Plant extracts represent a great source of molecules, with insecticidal activity, which are used for pest control in several crop production systems. This work aimed to evaluate the toxicity of an aqueous extract of leaves of castor bean against larvae of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) in search for different classes of molecules with insecticidal activities by using in vitro assays. The effects of the castor bean leaf extract on the food utilization, development, and survival of S. frugiperda larvae was evaluated by feeding the larvae an artificial diet supplemented with different concentrations of the extract (0%, 1%, 2.5%, 5%, and 10% w/v). The effects observed were dose-dependent, and the highest concentration evaluated (10% w/v) was the one the most affected food utilization by altering the nutritional indices, as well as larval weight gain, development time, and survivorship. In vitro assays to detect saponins, lectins, and trypsin inhibitors in the castor bean leaf extract were performed, but only trypsin inhibitors were detected. No preference for the diet source was detected in S. frugiperda by feeding the larvae in choice experiments with diets containing different concentrations of the castor bean extract tested. The data obtained indicate the existence of a potential molecule in the tested extract of castor bean to be used as an alternative insecticide to be integrated in the management of S. frugiperda.

THERMOSTABILITY OF XYLANOLYTIC ENZYMES PRODUCED BY LENTINULA EDODES UFV70

Xylanolytic enzymes produced by Lentinula edodes UFV70, cultivated in eucalyptus sawdust/rice bran medium, were stable at 50, 60 and 65 degrees C for 21 hours, losing only 15-25% activity. Fungus incubation at 50 degrees C for 12 hours and at 65 degrees C for 24 hours increased the amount of xylose produced.