Down-regulation of human papillomavirus E6 oncogene and antiproliferative effect of Schisandra chinensis and Pueraria lobata natural extracts on Hela cell line.

ETHNOPHARMACOLOGICAL RELEVANCE: Cervical cancer is one of the most common malignancies in women that continues to be a public health problem worldwide. Human papillomavirus (HPV) infection is closely related as the causative agent of almost all cases of cervical cancer. Currently, there is no effective treatment for the persistence of HPV. Although vaccines have shown promising results in recent years, they are still a costly strategy for developing countries and have no therapeutic effect on existing infections, which is why the need arises to search for new strategies that can be used in treatment, suppressing oncogenic HPV and disease progression. Extracts of Schisandra Chinensis and Pueraria lobata have been used in traditional medicine, and it has been shown in recent years that some of their bioactive compounds have pharmacological, antioxidant, antitumor, apoptotic, and proliferation effects in HPV-positive cells. However, its mechanism of action has yet to be fully explored. AIM OF THE STUDY: The following study aimed to determine the chemical composition, antioxidant activity, and potential antiproliferative and viral oncogene effects of natural extracts of S. chinensis and P. lobata on HPV-18 positive cervical cancer cells. MATERIALS AND METHODS: The HPV-18-positive HeLa cells were treated for 24 and 48 h with the ethanolic extracts of S chinensis and P. lobata. Subsequently, cell viability was evaluated using the resazurin method, the effect on the cell cycle of the extracts (1.0, 10, and 100 µg/mL) was measured by flow cytometry, the gene of expression of the E6/E7, P53, BCL-2, and E2F-1 were determined by RT-PCR and the protein expression of p53, Ki-67, x|and Bcl-2 by immunohistochemistry. Additionally, the chemical characterization of the two extracts was carried out using LC-MS, and the total phenolics content (TPC), Total flavonoid content (TFC), and DPPH radical scavenging capacity were determined. Data were analyzed using the Mann-Whitney and Kruskal Wallis U test with GraphPad Prism 6 software. RESULTS: The natural extracts of Schisandra chinensis and Pueraria lobata induced down-regulation of E6 HPV oncogene (p<0.05) and a strong up-regulation of P53 (p<0.05), E2F-1 (p<0.05), and Bcl-2 (p<0.05) gene expression. Simultaneously, the natural extracts tend to increase the p53 protein levels and arrest the cell cycle of HeLa in the G1/S phase (p<0.05). Investigated extracts were characterized by the occurrence of bioactive lignans and isoflavones in S. chinensis and P. lobata, respectively. CONCLUSION: The extracts of S. chinensis and P. lobata within their chemical characterization mainly present lignan and isoflavone-type compounds, which are probably responsible for inhibiting the expression of the HPV E6 oncogene and inducing an increase in the expression of p53, Bcl -2 and E2F-1 producing cell cycle detection in S phase in HeLa cells. Therefore, these extracts are good candidates to continue studying their antiviral and antiproliferative potential in cells transformed by HPV.

Effects of pueraria lobata combined with high-intensity intermittent training on skeletal muscle quality and gene expression / Efeitos da pueraria lobata combinados com treino intermitente de alta intensidade na qualidade músculoesquelética e expressão genética / Efecto de pueraria lobata y entrenamiento intermitente de alta intensidad sobre la calidad del músculo esquelético y la expresión génica

ABSTRACT Extraction of effective components from Pueraria lobata has important value for skeletal muscle quality and gene expression. The improvement effect of traditional high-intensity intermittent training on skeletal muscle has not been obvious, and it is difficult to guarantee the properties of some volatiles. Based on this, this paper analyzes the effect of high-intensity intermittent training on skeletal muscle quality and gene expression in Pueraria lobata. Based on a brief summary of extraction of Pueraria lobata, status of research on the pharmaceutical components of Pueraria lobata was summarized. Different specimens of Pueraria lobata were selected as research objects, and the process of high-intensity intermittent training was designed. High-intensity intermittent training, solvent extraction and water solvent extraction were combined together to design the fixed-bed continuous extraction scheme. According to the influence of Pueraria lobata on skeletal muscle quality, the influence of intermittent training on skeletal muscle quality was analyzed. The extraction results showed that Pueraria lobata combined with high-intensity intermittent training can effectively improve the content of skeletal muscle and ensure the effective expression of skeletal muscle gene.

RESUMO A extração de componentes eficazes da Pueraria lobata tem importante valor para a qualidade músculoesquelética e para a expressão genética. O efeito da melhoria do tradicional treinamento intervalado de alta intensidade na estrutura músculoesquelética não tem sido óbvio, e é difícil garantir as propriedades de alguns voláteis. Com base nisso, este estudo analisa o efeito do treinamento intervalado de alta intensidade na qualidade músculoesquelética e na expressão genética na Pueraria lobata. Com base num breve resumo da extração da Pueraria lobata, resumiu-se o andamento das pesquisas sobre os componentes farmacêuticos da Pueraria lobata. Diferentes amostras de Pueraria lobata foram selecionadas como objeto de pesquisa, e formulou-se o processo do treinamento intervalado de alta intensidade. O treinamento intervalado de alta intensidade, a extração de solventes e a extração de solventes à base de água foram combinadas para conceber o sistema de extração contínua de leito fixo. De acordo com a influência da Pueraria lobata na qualidade músculoesquelética, analisou-se a influência do treino intervalado na qualidade músculoesquelética. Os resultados da extração mostraram que a Pueraria lobata, combinada com treino intervalado de alta intensidade, pode melhorar, de maneira eficaz, o teor músculoesquelético e garantir a expressão eficaz da expressão genética do músculoesquelético.

RESUMEN La extracción de componentes eficaces de la Pueraria lobata tiene un importante valor para la calidad músculoesquelética y para la expresión genética. El efecto de la mejora del tradicional entrenamiento intercalado de alta intensidad en la estructura músculoesquelética no ha sido obvio, y es difícil garantizar las propriedades de algunos volátiles. Basándose en eso, este estudio analiza el efecto del entrenamiento intercalado de alta intensidad en la calidad músculoesquelética y en la expresión genética en la Pueraria lobata. Basándose en un breve resumen de la extracción de la Pueraria lobata, se resumió el andamiento de las investigaciones sobre los componentes farmacéuticos de la Pueraria lobata. Diferentes muestras de Pueraria lobata fueron seleccionadas como objeto de investigación, y se formuló el proceso del entrenamiento intercalado de alta intensidad. El entrenamiento intercalado de alta intensidad, la extracción de solventes y la extracción de solventes a base de agua fueron combinadas para concebir el sistema de extracción continua de lecho fijo. De acuerdo con la influencia de la Pueraria lobata en la calidad músculoesquelética, se analizó la influencia del entrenamiento intercalado en la calidad músculoesquelética. Los resultados de la extracción mostraron que la Pueraria lobata, combinada con entrenamiento intercalado de alta intensidad, puede mejorar, de manera eficaz, el tenor músculoesquelético y garantizar la expresión eficaz de la expresión genética del músculoesquelético.

Micronutrients content in Pueraria phaseoloides L. on soils under oil palm plantations in Tailândia, state of Pará / Acúmulo de micronutrientes em Kudzu tropical (Pueraria phaseoloides L) em solos sob plantações de dendê em Tailândia ­ Pará

The adoption of cover crops in commercial plantations has been shown to be an alternative to promote greater soil protection, enhancement in its structure and aggregation, optimization of accumulation and nutrient cycling to obtain greater production of biomass and higher nitrogen fixation. To determine the accumulation of micronutrients of Pueraria phaseoloides L., according to age, established as a soil cover in industrial oil palm crops, an experiment was carried out at Companhia Agroindustrial (CRAI) in the municipality of Tailândia, State of Pará, in an Oxisol. The experimental design was a randomized block with four replicates. The results allow to conclude that the accumulation of nutrients, except for Mn and Cu in the live Pueraria phaseoloides L. plant cover reduced over the years and those of B, Mn, Zn, and Cu were greater in the dead Pueraria phaseoloides L. plant cover and for Cl it was in the live Pueraria phaseoloides L. plant cover. The decreasing order of total micronutrient extraction was the following: Cl> Mn> B> Zn> Cu.

A adoção de plantas de cobertura em plantios comerciais tem se mostrado uma alternativa para promover ao solo maior proteção, melhorias em sua estrutura e agregação e otimização de acúmulo e ciclagem de nutrientes visando maior produção de biomassa e maior fixação de N2. Com o objetivo de determinar o acúmulo de micronutrientes puerária (Pueraria phaseoloides L.), em função da idade, estabelecida como cobertura de solo em plantações industriais de dendezeiros, instalou-se experimento na Companhia Real Agroindustrial (CRAI) no município de Tailândia, Estado do Pará, em Latossolo Amarelo. O delineamento experimental foi o de blocos ao acaso com quatro repetições. Os resultados permitem concluir que: o acúmulo de nutrientes com exceção do Mn e Cu na cobertura verde de Pueraria phaseoloides L., reduziu com o decorrer dos anos; os de B, Mn, Zn e Cu foram maiores na cobertura morta, enquanto que o de Cl na cobertura verde. A ordem decrescente da extração total de micronutrientes foi: Cl > Mn > B > Zn > Cu.

Tropical kudzu as a protein source in sheep diets / Puerária como fonte proteica em dietas de ovinos

Protein nutrition plays a key role in ruminant performance and represents the largest proportion of costs in animal feed. The present study evaluated whether the roughage portion of Santa Inês sheep diets could be supplemented with different ratios of tropical kudzu as a protein source and assessed the best inclusion level of tropical kudzu based on digestibility, intake, nitrogen balance and feeding behavior. Twenty rams were evaluated during the 20-day experimental period, and they were fed formulated diets with a roughage portion (60%) consisting of Elephant grass and tropical kudzu, the latter supplemented at different concentrations (0, 25, 50 and 75% on a dry matter basis). Forty percent of the concentrate portion consisted primarily of ground corn and soybean meal adjusted to obtain an isoproteic diet. Chemical and bromatological analyses, as well as, analyses to assess intake, digestibility, nitrogen balance and feeding behavior, were conducted. No statistically significant differences were found (P>0.05) for the intake and digestibility of dry matter (DM), crude protein (CP), ether extract (EE), neutral detergent fiber (NDF), hemicellulose (HEMI), non-fiber carbohydrates (NFC) and total digestible nutrients (TDN) or for the feeding behavior (feeding, rumination and idle times in h/day) and nitrogen balance between the diets studied. Thus, tropical kudzu can be considered a feed alternative to supplementation with prime protein foods (in this case, soybean meal) in sheep diets.(AU)

Objetivou-se com o presente estudo avaliar a inclusão na porção volumosa de diferentes porções de puerária como fonte proteica em dietas de ovinos da Raça Santa Inês, determinando a melhor concentração de inclusão de puerária com base na digestibilidade, consumo, balanço de nitrogênio e comportamento ingestivo. Durante os 20 dias do período experimental, foram utilizados 20 ovinos, que receberam dietas formuladas, cuja porção volumosa (60%) foi constituída por capim-Elefante e puerária, incluída em diferentes concentrações (0; 25; 50 e 75% com base na matéria seca). Os 40% da porção concentrada foram constituidos a base de milho moído e farelo de soja ajustados para obtenção de dietas isoprotéicas. Foram realizadas análises químico-bromatológicas, bem como análises para determinação de consumo, digestibilidade, balanço de nitrogênio e análise do comportamento ingestivo. Não foram observadas diferenças (p>0,05) no consumo e digestibilidade de MS; PB, EE, FDN, HEMI, CNF e nos valores de NDT, bem como no comportamento ingestivo (tempos de alimentação, ruminação e ócio em h/dia) e balanço de nitrogênio entre as dietas estudadas. A puerária é uma alternativa de alimento à inclusão de alimentos protéicos nobres (no caso, farelo de soja) em dietas de ovinos.(AU)

Tropical kudzu as a protein source in sheep diets / Puerária como fonte proteica em dietas de ovinos

Protein nutrition plays a key role in ruminant performance and represents the largest proportion of costs in animal feed. The present study evaluated whether the roughage portion of Santa Inês sheep diets could be supplemented with different ratios of tropical kudzu as a protein source and assessed the best inclusion level of tropical kudzu based on digestibility, intake, nitrogen balance and feeding behavior. Twenty rams were evaluated during the 20-day experimental period, and they were fed formulated diets with a roughage portion (60%) consisting of Elephant grass and tropical kudzu, the latter supplemented at different concentrations (0, 25, 50 and 75% on a dry matter basis). Forty percent of the concentrate portion consisted primarily of ground corn and soybean meal adjusted to obtain an isoproteic diet. Chemical and bromatological analyses, as well as, analyses to assess intake, digestibility, nitrogen balance and feeding behavior, were conducted. No statistically significant differences were found (P>0.05) for the intake and digestibility of dry matter (DM), crude protein (CP), ether extract (EE), neutral detergent fiber (NDF), hemicellulose (HEMI), non-fiber carbohydrates (NFC) and total digestible nutrients (TDN) or for the feeding behavior (feeding, rumination and idle times in h/day) and nitrogen balance between the diets studied. Thus, tropical kudzu can be considered a feed alternative to supplementation with prime protein foods (in this case, soybean meal) in sheep diets.

Objetivou-se com o presente estudo avaliar a inclusão na porção volumosa de diferentes porções de puerária como fonte proteica em dietas de ovinos da Raça Santa Inês, determinando a melhor concentração de inclusão de puerária com base na digestibilidade, consumo, balanço de nitrogênio e comportamento ingestivo. Durante os 20 dias do período experimental, foram utilizados 20 ovinos, que receberam dietas formuladas, cuja porção volumosa (60%) foi constituída por capim-Elefante e puerária, incluída em diferentes concentrações (0; 25; 50 e 75% com base na matéria seca). Os 40% da porção concentrada foram constituidos a base de milho moído e farelo de soja ajustados para obtenção de dietas isoprotéicas. Foram realizadas análises químico-bromatológicas, bem como análises para determinação de consumo, digestibilidade, balanço de nitrogênio e análise do comportamento ingestivo. Não foram observadas diferenças (p>0,05) no consumo e digestibilidade de MS; PB, EE, FDN, HEMI, CNF e nos valores de NDT, bem como no comportamento ingestivo (tempos de alimentação, ruminação e ócio em h/dia) e balanço de nitrogênio entre as dietas estudadas. A puerária é uma alternativa de alimento à inclusão de alimentos protéicos nobres (no caso, farelo de soja) em dietas de ovinos.

Frequências de cortes em nove leguminosas forrageiras tropicais herbáceas cultivadas ao sol e sob plantação florestal / Cutting frequencies of nine herbaceous tropical forage legumes cultivated under full sun light and under a forest plantation

O objetivo deste trabalho foi estudar a produção e o valor nutritivo de nove espécies de leguminosas forrageiras herbáceas tropicais, cultivadas sob dois níveis de irradiação luminosa e seis frequências de corte na estação chuvosa, em Anhembi (SP), Brasil. Parcelas puras foram estabelecidas ao sol e sob plantação de Eucalyptus grandis, com 8 anos de idade, com 1.000 árvores/ha, que interceptou mais de 80% da radiação luminosa incidente. Avaliou-se a taxa de acúmulo de matéria seca, a proteína bruta e a digestibilidade in vitro da matéria seca em amostras de plantas colhidas em intervalos de 30, 60, 90 e 180 dias, com cortes adicionais aos 120 e 150 dias. Sob pleno sol, as leguminosas testadas toleraram cortes mais frequentes (intervalos de 30 - 90 dias), exceto o Lablab. Na sombra, o grau de tolerância variou conforme a espécie; em termos de produção de MS e de persistência, a Centrosema e a Puerária se destacaram dentre as demais leguminosas comparadas, quando os cortes foram realizados em intervalos de 60 dias, enquanto o Arachis e o Estilosantes foram os mais produtivos quando os cortes ocorreram em intervalos de 120 dias, mantendo o valor nutricional. O Alysicarpus e a Aeschynomene não persistiram sob o nível de sombreamento que prevaleceu nesse experimento.

The aim of this study was to evaluate the production and nutritive value of nine tropical legumes under two light intensities and six cutting regimens in the wet season, in Anhembi (SP) - Brazil. Pure stands were established in full light and under an eight-year-old eucalyptus plantation with 1.000 plants per hectare. The trees intercepted over80% of the incident radiation. Dry matter herbage accumulation, dry matter in vitro digestibility and crude protein under the cutting intervals 30, 60, 90 and 180 days, with additional cuttings at 120 and 150 days were evaluated. With the exception of Lablab, all other tested legume species tolerated higher cutting frequency under when grown under full sun light then under shade. The degree of shade-tolerance varied according to the species; Centrosema and Pueraria produced well at cutting intervals of 60 days, showing higher dry matter production and persistence while Arachis and Stylosanthes showed better results under longer cutting intervals, keeping the nutritional value, and can be indicated for differential grazing. Alysicarpus and Aeschynomene did not persist under the prevailing conditions.

Frequências de cortes em nove leguminosas forrageiras tropicais herbáceas cultivadas ao sol e sob plantação florestal / Cutting frequencies of nine herbaceous tropical forage legumes cultivated under full sun light and under a forest plantation

O objetivo deste trabalho foi estudar a produção e o valor nutritivo de nove espécies de leguminosas forrageiras herbáceas tropicais, cultivadas sob dois níveis de irradiação luminosa e seis frequências de corte na estação chuvosa, em Anhembi (SP), Brasil. Parcelas puras foram estabelecidas ao sol e sob plantação de Eucalyptus grandis, com 8 anos de idade, com 1.000 árvores/ha, que interceptou mais de 80% da radiação luminosa incidente. Avaliou-se a taxa de acúmulo de matéria seca, a proteína bruta e a digestibilidade in vitro da matéria seca em amostras de plantas colhidas em intervalos de 30, 60, 90 e 180 dias, com cortes adicionais aos 120 e 150 dias. Sob pleno sol, as leguminosas testadas toleraram cortes mais frequentes (intervalos de 30 - 90 dias), exceto o Lablab. Na sombra, o grau de tolerância variou conforme a espécie; em termos de produção de MS e de persistência, a Centrosema e a Puerária se destacaram dentre as demais leguminosas comparadas, quando os cortes foram realizados em intervalos de 60 dias, enquanto o Arachis e o Estilosantes foram os mais produtivos quando os cortes ocorreram em intervalos de 120 dias, mantendo o valor nutricional. O Alysicarpus e a Aeschynomene não persistiram sob o nível de sombreamento que prevaleceu nesse experimento(AU)

The aim of this study was to evaluate the production and nutritive value of nine tropical legumes under two light intensities and six cutting regimens in the wet season, in Anhembi (SP) - Brazil. Pure stands were established in full light and under an eight-year-old eucalyptus plantation with 1.000 plants per hectare. The trees intercepted over80% of the incident radiation. Dry matter herbage accumulation, dry matter in vitro digestibility and crude protein under the cutting intervals 30, 60, 90 and 180 days, with additional cuttings at 120 and 150 days were evaluated. With the exception of Lablab, all other tested legume species tolerated higher cutting frequency under when grown under full sun light then under shade. The degree of shade-tolerance varied according to the species; Centrosema and Pueraria produced well at cutting intervals of 60 days, showing higher dry matter production and persistence while Arachis and Stylosanthes showed better results under longer cutting intervals, keeping the nutritional value, and can be indicated for differential grazing. Alysicarpus and Aeschynomene did not persist under the prevailing conditions(AU)

Dispersão e persistência de leguminosas forrageiras tropicais após ingestão por bovinos / Dispersion and persistence of leguminous tropical forage after ingestion by cattle

Entre os fatores determinantes da baixa utilização do consórcio gramínea-leguminosa, pode ser citado a implantação das leguminosas em pastagens já estabelecida. No entanto, animais podem servir como mecanismo de dispersão das sementes. Assim, objetivou-se neste estudo avaliar a introdução das leguminosas calopogônio, macrotiloma, kudzu tropical e soja perene em pastagem estabelecida de Brachiaria decumbens, por meio da dispersão em fezes de bovinos. Para esta finalidade, foi utilizado cinco novilhas como veiculadoras das sementes, as quais foram ofertadas 180g de sementes de cada leguminosa misturado ao concentrado de forma alternada, esperando um período de 48h para completa defecação das sementes. Todas as leguminosas foram encontradas nas placas fecais, porém o kudzu tropical apresentou maior número de exemplares encontrados em relação às demais (P<0,05). Quando avaliada a persistência após estabelecimento e pastejo, foi encontrado menor número de exemplares para o kudzu tropical em relação às outras três leguminosas (P<0,05), que não diferiram entre si. No entanto, a porcentagem de sementes germinadas nas fezes em relação à quantidade de sementes ofertadas em 180g de sementes para este mecanismo de propagação pode ser considerado baixo para as quatro leguminosas estudadas.

Among the factors to the low utilization of grass-leguminous intercropping, may be cited the deployment of leguminous in pastures already established. However, animals can serve as a seed dispersal mechanism. Thus, this study aimed to evaluate the introduction of leguminous calopogonium, macrotiloma, tropical kudzu and soybeans, in perennial pastures of Brachiaria decumbens, through dispersal in cattle dung. For this purpose we used five heifers as seeds disseminator, which were offered 180g seeds of each leguminous, alternately mixed with the concentrated, waiting a period of 48 hours to complete the seed defecation. All leguminous were found in fecal plates, but kudzu had the greatest number of specimens found in relation to others (P <.05). However, when evaluated persistence after establishment and grazing found fewer copies to kudzu in the other three leguminous (P <0.05) that did not differ among themselves. When, the percentage of seeds germinated in the stool, relative to the amount offered in 180g of seed for this propagation mechanism, can be considered low for the four leguminous studied.

Phenotypic plasticity of Neonotonia wightii and Pueraria phaseoloides grown under different light intensities.

Plants have the ability to undergo morphophysiological changes based on availability of light. The present study evaluated biomass accumulation, leaf morphoanatomy and physiology of Neonotonia wightii and Pueraria phaseoloides grown in full sunlight, as well as in 30% and 50% shade. Two assays were performed, one for each species, using a randomized block design with 10 replicates. A higher accumulation of fresh mass in the shoot of the plants was observed for both species under cultivation in 50% shade, while no differences were detected between the full sunlight and 30% shade. N. wightii and P. phaseoloides showed increase in area and reduction in thickness leaf when cultivated in 50% shade. There were no changes in photosynthetic rate, stomatal conductance, water use efficiency and evapotranspiration of P. phaseoloides plants because growth environment. However, the shade treatments caused alterations in physiological parameters of N. wightii. In both species, structural changes in the mesophyll occurred depending on the availability of light; however, the amount of leaf blade tissue remained unaltered. Despite the influence of light intensity variation on the morphophysiological plasticity of N. wightii and P. phaseoloides, no effects on biomass accumulation were observed in response to light.

Adubação verde e alterações nas características químicas de um Cambissolo na região de Ji-Paraná-RO / Green manure and changes on chemical characteristics of a soil in the Ji-Paraná-RO region

A adubação verde promove benefícios nas características químicas, físicas e biológicas do solo. O trabalho avaliou o efeito da incorporação de diferentes adubos verdes nas características químicas de um Cambissolo háplico eutrófico. Foi utilizado o delineamento em blocos ao acaso, com oito tratamentos e três repetições. Os tratamentos consistiram das seguintes leguminosas: Calopogonium mucunoides, Crotalaria juncea, C. spectabilis, Cajanus cajan, Macrotyloma, Mucuna pruriens, Pueraria phaseoloides e a testemunha (sem leguminosa). As avaliações foram realizadas 60 dias após o corte e incorporação. A utilização de leguminosas como adubação verde proporcionou aumento nos teores de matéria orgânica, soma de bases e percentagem de saturação por bases, destacando-se a Pueraria phaseoloides, C. juncea e C. spectabilis.

The green manure promotes benefits on the chemical, physical and biological properties of soil. The experiment evaluated the effect of incorporation of differents green manure on the chemistry characteristics of a soil. Experimental design used was a randomized block, with eight treatments and three replications. Treatments were: Calopogonium mucunoides, Crotalaria juncea, Crotalaria spectabilis, Cajanus cajan, Macrotyloma axillare, Mucuna pruriens and Pueraria phaseoloides and the treatment control (without leguminous)). Evaluations of soil samplings were performed in 60 days after handling were performed. The results showed significant effects of the leguminous crop Pueraria phaseoloides, C. juncea and C. spectabilis on the fertility of the soil, with significant increments of organic matter, exchangeable bases and in the base saturation.

Differential binding with ERá and ERâ of the phytoestrogen-rich plant Pueraria mirifica

Variations in the estrogenic activity of the phytoestrogen-rich plant, Pueraria mirifica, were determined with yeast estrogen screen (YES) consisting of human estrogen receptors (hER) hERá and hERâ and human transcriptional intermediary factor 2 (hTIF2) or human steroid receptor coactivator 1 (hSRC1), respectively, together with the â-galactosidase expression cassette. Relative estrogenic potency was expressed by determining the â-galactosidase activity (EC50) of the tuber extracts in relation to 17â-estradiol. Twenty-four and 22 of the plant tuber ethanolic extracts interacted with hERá and hERâ, respectively, with a higher relative estrogenic potency with hERâ than with hERá. Antiestrogenic activity of the plant extracts was also determined by incubation of plant extracts with 17â-estradiol prior to YES assay. The plant extracts tested exhibited antiestrogenic activity. Both the estrogenic and the antiestrogenic activity of the tuber extracts were metabolically activated with the rat liver S9-fraction prior to the assay indicating the positive influence of liver enzymes. Correlation analysis between estrogenic potency and the five major isoflavonoid contents within the previously HPLC-analyzed tuberous samples namely puerarin, daidzin, genistin, daidzein, and genistein revealed a negative result.

Differential binding with ERalpha and ERbeta of the phytoestrogen-rich plant Pueraria mirifica.

Variations in the estrogenic activity of the phytoestrogen-rich plant, Pueraria mirifica, were determined with yeast estrogen screen (YES) consisting of human estrogen receptors (hER) hERalpha and hERbeta and human transcriptional intermediary factor 2 (hTIF2) or human steroid receptor coactivator 1 (hSRC1), respectively, together with the beta-galactosidase expression cassette. Relative estrogenic potency was expressed by determining the beta-galactosidase activity (EC(50)) of the tuber extracts in relation to 17beta-estradiol. Twenty-four and 22 of the plant tuber ethanolic extracts interacted with hERalpha and hERbeta, respectively, with a higher relative estrogenic potency with hERbeta than with hERalpha. Antiestrogenic activity of the plant extracts was also determined by incubation of plant extracts with 17beta-estradiol prior to YES assay. The plant extracts tested exhibited antiestrogenic activity. Both the estrogenic and the antiestrogenic activity of the tuber extracts were metabolically activated with the rat liver S9-fraction prior to the assay indicating the positive influence of liver enzymes. Correlation analysis between estrogenic potency and the five major isoflavonoid contents within the previously HPLC-analyzed tuberous samples namely puerarin, daidzin, genistin, daidzein, and genistein revealed a negative result.

The mutagenic and antimutagenic effects of the traditional phytoestrogen-rich herbs, Pueraria mirifica and Pueraria lobata.

Pueraria mirifica is a Thai phytoestrogen-rich herb traditionally used for the treatment of menopausal symptoms. Pueraria lobata is also a phytoestrogen-rich herb traditionally used in Japan, Korea and China for the treatment of hypertension and alcoholism. We evaluated the mutagenic and antimutagenic activity of the two plant extracts using the Ames test preincubation method plus or minus the rat liver mixture S9 for metabolic activation using Salmonella typhimurium strains TA98 and TA100 as indicator strains. The cytotoxicity of the two extracts to the two S. typhimurium indicators was evaluated before the mutagenic and antimutagenic tests. Both extracts at a final concentration of 2.5, 5, 10, or 20 mg/plate exhibited only mild cytotoxic effects. The plant extracts at the concentrations of 2.5, 5 and 10 mg/plate in the presence and absence of the S9 mixture were negative in the mutagenic Ames test. In contrast, both extracts were positive in the antimutagenic Ames test towards either one or both of the tested mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and benzo(a)pyrene. The absence of mutagenic and the presence of anti-mutagenic activities of the two plant extracts were confirmed in rec-assays and further supported by a micronucleus test where both plant extracts at doses up to 300 mg/kg body weight (equivalent to 16 g/kg body weight plant tuberous powder) failed to exhibit significant micronucleus formation in rats. The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.

The mutagenic and antimutagenic effects of the traditional phytoestrogen-rich herbs, Pueraria mirifica and Pueraria lobata

Pueraria mirifica is a Thai phytoestrogen-rich herb traditionally used for the treatment of menopausal symptoms. Pueraria lobata is also a phytoestrogen-rich herb traditionally used in Japan, Korea and China for the treatment of hypertension and alcoholism. We evaluated the mutagenic and antimutagenic activity of the two plant extracts using the Ames test preincubation method plus or minus the rat liver mixture S9 for metabolic activation using Salmonella typhimurium strains TA98 and TA100 as indicator strains. The cytotoxicity of the two extracts to the two S. typhimurium indicators was evaluated before the mutagenic and antimutagenic tests. Both extracts at a final concentration of 2.5, 5, 10, or 20 mg/plate exhibited only mild cytotoxic effects. The plant extracts at the concentrations of 2.5, 5 and 10 mg/plate in the presence and absence of the S9 mixture were negative in the mutagenic Ames test. In contrast, both extracts were positive in the antimutagenic Ames test towards either one or both of the tested mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and benzo(a)pyrene. The absence of mutagenic and the presence of anti-mutagenic activities of the two plant extracts were confirmed in rec-assays and further supported by a micronucleus test where both plant extracts at doses up to 300 mg/kg body weight (equivalent to 16 g/kg body weight plant tuberous powder) failed to exhibit significant micronucleus formation in rats. The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.

Production and economic potentials of cattle in pasture-based systems of the western Amazon region of Brazil.

Our objectives were to evaluate strategies to improve productivity and economic returns from beef and dual-purpose cattle systems based on data collected on one dual-purpose (Bos taurus x Bos indicus) and two beef (Nellore) cattle farms in the western Amazon region of Brazil. Forage chemical composition and digestion rates of carbohydrate fractions of grazed Brachiaria decumbens and Brachiaria brizantha cv. Marandu grasses and Pueraria phaseoloides (tropical kudzu) legume were measured monthly during a 9-mo period from the end of one dry season to the end of the subsequent rainy season. Measurements of milk and growth responses to grazing these forages were used to predict animal productivity responses to dietary nutrient availability throughout an annual cycle. The ME available for gain in our simulations was always more limiting than metabolizable protein. The predicted ME available for gain was 0.50 kg/d for steers grazing B. brizantha and 0.40 kg/d for finishing steers grazing B. decumbens. Grasses contained more NDF and neutral detergent insoluble protein and less ME (P < 0.05) in the rainiest months than in the less rainy season, which resulted in 20% less predicted weight gain by growing steers (P < 0.05). Supplementation with sorghum grain was required to increase milk production and growth by 25 or 50% per animal, respectively, but this strategy was less profitable than current forage-only diets. Greater productivity of land and labor from higher stocking indicated greater net margins for beef production, but not for milk. This study suggested that more intensive beef production by judicious fertilization of grass-legume pastures and greater stocking density is the preferable strategy for owners of these cattle systems to improve economic returns under current conditions. It also might help decrease the motivation for additional forest clearing.