Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum / Composição química e efeito inibitório in vitro dos óleos essenciais das cascas dos frutos de três espécies de Citrus e do limoneno sobre o crescimento micelial de Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 μL. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 μL) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 μL doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum.

Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 µL. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 µL) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 µL doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Determinação da energia metabolizável do farelo residual do milho com e sem enzima em dietas para frangos de corte / Determination of the metabolizable energy of residual corn bran with and without enzyme in diets for broilers

O objetivo deste trabalho foi avaliar o valor nutricional e determinar a energia metabolizável do farelo residual de milho (FRM) sem e com o uso da enzima alfa- amilase. Foi realizado um experimento de metabolismo com 180 pintos machos Cobb com 14 dias, distribuídos em um delineamento inteiramente ao acaso, com seis tratamentos, cinco repetições e seis aves por parcela. As dietas experimentais foram: T1: ração referência (RR), T2: 60% T1 + 40% de FRM, T3: RR + enzima, T4: 60% T1 + 40% de FRM com adição de enzima, T5: RR com substituição de 100% do milho pelo FRM e T6: RR com substituição de 100% do milho pelo FRM com adição de enzima. A composição química do FRM foi: 88,33% de matéria seca (MS), 10,23% de proteína bruta (PB), 15,44% de extrato etéreo (EE), 4,33% de cinzas (CZ) e 4555kcal/kg de energia bruta (EB). Os valores dos coeficientes de metabolizabilidade aparente para o FRM sem e com adição de enzima foram, respectivamente, de 73,37% e 76,33% para MS (p=0,0136), 70,44% e 70,39% para PB (p=0,9595) e de 74,79% e 76,77% para EB (p=0,0128). Os valores da energia metabolizável aparente (EMA) e da EMA corrigida para retenção de nitrogênio (EMAn) para o FRM (na base natural) foram de 3322±19 e 3241±18kcal/kg e de 3334±16 e 3261±17kcal/kg, respectivamente, sem e com adição de enzima. A adição da enzima não teve efeito estatístico significativo sobre os valores de EMA e EMAn, entretanto houve efeito positivo no coeficiente de metabolização da energia.(AU)

The aim of this study was to evaluate the nutritional value and the metabolizable energy of residual corn bran (FRM) with and without the use of alpha amylase enzyme. One metabolism assay was done using 180 Ross male broilers at 14 days of age distributed in an entirely randomized design with six treatments, five replications and six broilers per plot. The assay treatments established were: T1: reference diet (RR), T2: 60% T1 + 40% of FRM, T3: T1 (RR) + enzyme addition, T4: 60% T1 + 40% of FRM with enzyme addition, T5: T1 (RR) with 100% replacement of corn by FRM and T6: T1 (RR) with 100% replacement of corn by FRM with enzyme addition. FRM chemical composition was: 88.33% of dry matter (MS), 10.23% of crude protein (PB), 15.44% of crude fat (EE), 4.33% of ashes (CZ) and 4555Kcal/kg of Gross Energy (EB). The FRM apparent metabolizability coefficients with and without the enzyme addition were, respectively, 73.37% and 76.33% for MS (p=0.0136), 70.44% and 70.39% for PB (p=0.9595) and 74.79% and 76.77% for EB (p=0.0128). The apparent metabolizable energy (EMA) and EMA corrected for nitrogen retention values (as is basis) were 3322±19 and 3241±18Kcal/kg and 3334±16 and 3261±17Kcal/kg, respectively, for with and without the enzyme addition to FRM. The enzyme addition had no statistical significance on the EMA and EMAn values, however, a significant effect on energy metabolizability coefficient was observed.(AU)

Avaliação das atividades antibacteriana, tripanocida e citotóxica do extrato hidroalcóolico das raízes de Tradescantia sillamontana Matuda (Veludo Branco) (Commelinaceae) / Evaluation of antibacterial, trypanocidal and cytotoxical activities of the hydroalcoholic extract from the roots of Tradescantia sillamontana Matuda (Veludo Branco) (Commelinaceae)

RESUMO O objetivo deste estudo foi avaliar o efeito antibacteriano e tripanocida in vitro do extrato hidroalcóolico das raízes de Tradescantia sillamontana Matuda (Commelinaceae), conhecida popularmente como veludo branco. Foi avaliada a atividade antibacteriana in vitro frente às bactérias Streptococcus mitis (CIM = 100 µg/mL; CMB = 150 µg/mL), Streptococcus mutans (CIM = 200 µg/mL; CMB = 220 µg/mL), Streptococcus sanguinis (CIM = 400 µg/mL; CMB = 425 µg/mL), Streptococcus sobrinus (CIM = 400 µg/mL; CMB = 420 µg/mL) e Bacteroides fragilis (CIM = 400 µg/mL; CMB = 430 µg/mL) pelo método de diluição em caldo. Os protozoários da família tripanossomatídeo causam doenças tropicais que costumam ser negligenciadas que costumam ser como a tripanossomíase, para a qual estão disponíveis poucos medicamentos. Neste contexto, o extrato hidroalcóolico das raízes de T. sillamontana também foi avaliado frente às formas tripomastigotas da cepa Y de Trypanosoma cruzi, com promissora atividade frente a este protozoário (IC50 = 2,4 µg/mL). Quando avaliada a atividade citotóxica frente a fibroblastos da linhagem LLCMK2, o extrato apresentou moderada citotoxicidade (CC50 = 480,37 µg/mL). Os resultados ora apresentados para o extrato hidroalcóolico das raízes de Tradescantia sillamontana Matuda demonstraram promissoras atividades antibacteriana e tripanocida, sendo uma fonte alternativa de produtos naturais com atividades contra T. cruzi e algumas bactérias do gênero Streptococcus e Bacteroides.

ABSTRACT The aim of this study was to investigate the in vitro, antibacterial and trypanocidal effect of the hydroalcoholic extract from the roots of Tradescantia sillamontana Matuda (Commelinaceae), commonly known as Veludo branco. The in vitro antibacterial activity against the standard bacteria Streptococcus mitis (MIC = 100µg/mL; MBC = 150 µg/mL), Streptococcus mutans (MIC = 200µg/mL; MBC = 220 µg/mL), Streptococcus sanguinis (MIC = 400µg/mL; MBC = 425 µg/mL), Streptococcus sobrinus (MIC = 400µg/mL; MBC = 420 µg/mL) andBacteroides fragilis (MIC = 400µg/mL; MBC = 430 µg/mL), using microdilution broth methods. Protozoans from the trypanosomatid family cause neglected tropical diseases such as trypanosomiasis, for which few drugs are available. In this context, the hydroalcoholic extract of the Tradescantia sillamontana roots was also investigated with regards to the in vitro effects against the trypomastigote forms of theY strain of Trypanosoma cruzi, showing strong activity against this parasite (IC50 = 2.4 µg/mL). When performing cytotoxic activity against fibroblasts LLCMK2 line, the extract showed moderate cytotoxicity (CC50 = 480.37 mg/mL). The results presented for the hydroalcoholic extract of the roots of Tradescantia sillamontana Matuda demonstrated effective antibacterial and trypanocidal activities and were shown to be an alternative source of natural products with activity against T. cruzi and some bacteria of the genus Streptococcus and Bacteroides.

Value of sentinel lymph node biopsy in papillary thyroid cancer: initial results of a prospective trial.

The objectives of the study were to evaluate the performance of sentinel lymph node biopsy (SLNB) in detecting occult metastases in papillary thyroid carcinoma (PTC) and to correlate their presence to tumor and patient characteristics. Twenty-three clinically node-negative PTC patients (21 females, mean age 48.4 years) were prospectively enrolled. Patients were submitted to sentinel lymph node (SLN) lymphoscintigraphy prior to total thyroidectomy. Ultrasound-guided peritumoral injections of (99m)Tc-phytate (7.4 MBq) were performed. Cervical single-photon emission computed tomography and computed tomography (SPECT/CT) images were acquired 15 min after radiotracer injection and 2 h prior to surgery. Intra-operatively, SLNs were located with a gamma probe and removed along with non-SLNs located in the same neck compartment. Papillary thyroid carcinoma, SLNs and non-SLNs were submitted to histopathology analysis. Sentinel lymph nodes were located in levels: II in 34.7 % of patients; III in 26 %; IV in 30.4 %; V in 4.3 %; VI in 82.6 % and VII in 4.3 %. Metastases in the SLN were noted in seven patients (30.4 %), in non-SLN in three patients (13.1 %), and in the lateral compartments in 20 % of patients. There were significant associations between lymph node (LN) metastases and the presence of angio-lymphatic invasion (p = 0.04), extra-thyroid extension (p = 0.03) and tumor size (p = 0.003). No correlations were noted among LN metastases and patient age, gender, stimulated thyroglobulin levels, positive surgical margins, aggressive histology and multifocal lesions. Sentinel lymph node biopsy can detect occult metastases in PTC. The risk of a metastatic SLN was associated with extra-thyroid extension, larger tumors and angio-lymphatic invasion. This may help guide future neck dissection, patient surveillance and radioiodine therapy doses.

Composição físico-química e valores energéticos de farinhas de silagem de peixe para frangos de corte / Physicochemical composition and energy values of fish silage meal for broilers

Objetivou-se determinar a composição físico-química, os valores energéticos e os coeficientes de digestibilidade de quatro farinhas de silagem de peixe para frangos de corte. Foram produzidas quatro farinhas de silagem de peixe, utilizando-se o resíduo da filetagem de tilápias ensilado com diferentes fontes de carboidratos fermentáveis. Analisou-se a composição físico-química das silagens, e, em seguida, um ensaio de metabolismo com 180 pintos machos da linhagem Cobb de 14 a 25 dias de idade. Também foram avaliados o tempo de trânsito gastrintestinal das rações e o desempenho das aves nas gaiolas. Os animais foram distribuídos em delineamento inteiramente ao acaso, com cinco tratamentos, seis repetições e seis aves por unidade experimental. Os tratamentos consistiram de uma dieta referência e de quatro dietas teste compostas de 60 porcento da ração referência e 40 porcento do resíduo da filetagem de tilápia ensilado com diferentes fontes de carboidratos, sendo a farinha de silagem de peixe com o farelo de algaroba (SFA), com a farinha de varredura de mandioca (SFVM), com o farelo de milho (SFM) e com a casca da mandioca (SCM). A SFM obteve o maior teor de PB, 22,38 porcento, de EE, 27,35 porcento, e o maior tempo de trânsito, com 195,0min; a SCM apresentou o maior valor de MM, 11,12 porcento. Os valores de EMA e EMAn das farinhas de silagem de peixe não diferiram significativamente entre eles. O maior GP e a melhor CA foram apresentados pelos animais do tratamento SFM, e os piores GP e CA pelos frangos de corte alimentados com dietas contendo a SFVM. Com base na composição obtida, estas silagens de peixe têm potencialidade para serem utilizadas em dietas para frangos de corte...

The objective of this research was to determine the physical and chemical composition energy value and digestibility of four fish silage meal for broilers. Four flours were produced from fish silage using the residue of tilapia filleting ensiled together with different sources of fermentable carbohydrates. We analyzed the physical and chemical composition of silages and then a metabolism trial with 180 male Cobb chicks 14-25 days old, and also evaluated gastrintestinal transit time of feed and performance of birds in cages. The animals were distributed in a completely randomized design with five treatments and six replicates of six birds each. Treatments consisted of a reference diet and four test diets composed of 60 percent of the reference diet with the inclusion of 40 percent of the residue of tilapia silage with different sources of carbohydrates, and the fish silage meal with bran mesquite (SFA) with the scan cassava flour (SFVM) with corn meal (SFM), and peel cassava (SCM). The SFM had the highest content of CP, 22.38 percent, EE, 27.35 percent, and the largest transit time with 195.0 min, the SCM showed the highest MM, 11.12 percent. The AME and AMEn of flour fish silage did not differ significantly between them. The biggest and best GP CA was presented by animals in the SFM treatment and the worse CA GP was for broilers fed diets containing SFVM. Based on the composition obtained, these fish silage have the potential to be used in diets for broilers...

The epimer of kaurenoic acid from Croton antisyphiliticus is cytotoxic toward B-16 and HeLa tumor cells through apoptosis induction.

Cancer has become the leading cause of death in developing countries due to increased life expectancy of the population and changes in lifestyle. Studies on active principles of plant have motivated researchers to develop new antitumor agents that are specific and effective for treatment of neoplasms. Kaurane diterpenes are considered important compounds in the development of new and highly effective anticancer chemotherapeutic agents due to their cytotoxic properties in the induction of apoptosis. We evaluated the cytotoxic and apoptotic activity of the epimer of kaurenoic acid (EKA) isolated from the medicinal plant Croton antisyphiliticus (Euphorbiaceae) toward tumor cell lines HeLa and B-16 and normal fibroblasts 3T3. Based on analyses with the MTT test, EKA showed cytotoxic activity, with half maximal inhibitory concentration values of 59.41, 68.18 and 60.30 µg/mL for the B-16, HeLa and 3T3 cell lines, respectively. The assay for necrotic or apoptotic cells by differential staining showed induction of apoptosis in all three cell lines. We conclude that EKA is not selective between tumor and normal cell lines; the mechanism of action of EKA is induction of apoptosis, which is part of the innate mechanism of cell defense against neoplasia.

Neutralization of pharmacological and toxic activities of Bothrops jararacussu snake venom and isolated myotoxins by Serjania erecta methanolic extract and its fractions

Most of the snakebites recorded in Brazil are caused by the Bothrops genus. Given that the local tissue damage caused by this genus cannot be treated by antivenom therapy, numerous studies are focusing on supplementary alternatives, such as the use of medicinal plants. Serjania erecta has already demonstrated anti-inflammatory, antiseptic and healing properties. In the current study, the aerial parts of S. erecta were extracted with methanol, then submitted to chromatographic fractionation on a Sephadex LH20 column and eluted with methanol, which resulted in four main fractions. The crude extract and fractions neutralized the toxic activities of Bothrops jararacussu snake venom and isolated myotoxins (BthTX-I and II). Results showed that phospholipase A2, fibrinogenolytic, myotoxic and hemorrhagic activities were inhibited by the extract. Moreover, the myotoxic and edematous activities induced by BthTX-I, and phospholipase A2 activity induced by BthTX-II, were inhibited by the extract of S. erecta and its fraction. The clotting time on bovine plasma was significantly prolonged by the inhibitory action of fractions SF3 and SF4. This extract is a promising source of natural inhibitors, such as flavonoids and tannins, which act by forming complexes with metal ions and proteins, inhibiting the action of serineproteases, metalloproteases and phospholipases A2.

Atividade alelopática de extratos aquosos de plantas medicinais na germinação de Lactuca sativa L. e Bidens pilosa L. / Allelopathic activity of aqueous extracts of medicinal plants on the germination of Lactuca sativa L. and Bidens pilosa L.

O potencial alelopático de quatro diferentes extratos de plantas medicinais (Eclipta alba, Gomphrena globosa, Tabernaemontana catharinensis e Tithonia diversifolia) foi avaliado na germinação de Bidens pilosa e Lactuca sativa. Dentre os resultados obtidos no teste de germinação, notou-se um maior poder inibitório do extrato de G. globosa sobre sementes e plântulas de Bidens pilosa e Lactuca sativa independente da concentração utilizada e aumento da atividade inibitória de extratos T. catharinensis e de T. diversifolia com aumento da concentração destes. Quanto ao Índice de Velocidade de Germinação, os dados demonstraram redução do índice devido a elevada mortalidade das plântulas submetidas aos extratos de G. globosa e de T. catharinensis e a germinação tardia das sementes submetidas ao extrato de T. diversifolia. Quanto ao vigor das sementes, as menores porcentagens de sobrevivência ocorreram extratos à 2,5 por cento para B. pilosa e 10 por cento para L. sativa.

The allelopathic potential of four different extracts of medicinal plants (Eclipta alba, Gomphrena globosa, Tabernaemontana catharinensis and Tithonia diversifolia) was evaluated on the germination of Bidens pilosa and Lactuca sativa. The results obtained in the germination test indicated a greater inhibitory activity of G. globosa extract on Bidens pilosa and Lactuca sativa seeds and seedlings, regardlessof the used concentration, and an increase in the inhibitory activity of T. catharinensis and T. diversifolia extracts with an increase in their concentrations. As to Germination Speed Index, the data demonstrated a reduction due to the high mortality of seedlings subjected to G. globosa and T. catharinensis extracts and a late germination of seeds subjected to T. diversifolia extracts. As to the vigor of seeds, the smallest survival percentages occur at 2.5 percent for B. pilosa and at 10 percent for L.sativa.

Antiophidian properties of plant extracts against Lachesis muta venom

Snakebites comprise a serious health problem in several countries due to their global incidence, which exceeds 2.5 million per year, and the elevated number of victim fatalities. To counteract envenomations, antivenoms have been used regularly for more than a century. Apart from side effects including anaphylactic reactions, antivenoms are not able to efficiently neutralize local tissue damage, which contributes to increasing the severity and morbidity observed in patients. This fact, in turn, may be responsible for economic hardship, particularly in rural populations of developing countries. In the present work, we evaluated the antiophidian properties of 12 Brazilian plant extracts against the hemolytic, coagulant, hemorrhagic and proteolytic effects of Lachesis muta venom. Taken together, our data revealed that most of these aqueous products were capable of inhibiting those activities at different levels, except for Sapindus saponaria extract. In contrast, Stryphnodendron barbatiman extract completely neutralized all the analyzed biological activities. Thus, we may conclude that Brazilian flora may also be useful against L. muta accidents.

Anti-inflammatory and anti-nociceptive effects of Zeyheria montana (Bignoniaceae) ethanol extract

In this work, the analgesic and anti-inflammatory activities of Zeyheria montana Mart. ethanol leaf extract were investigated at doses of 75, 150 and 300 mg/kg body weight. In the analgesic assay, against a chemical stimulus in mice, acetic acid-induced writhes were significantly inhibited by the extract at doses of 75 mg/kg (67.27 percent), 150 mg/kg (49.38 percent) and 300 mg/kg (82.87 percent). Also, a vigorous decrease in hyperalgesia was observed when measured after 2 h and 6 h of lipopolysaccharide stimulation of rats for all doses of extract tested. Z. montana extract, at doses of 75 and 300 mg/kg, caused very slight central analgesia in rats submitted to thermal stimulus, particularly noticeable at 30 min following treatment. The anti-inflammatory activity of Z. montana extract on carrageenan-induced oedema in rats was evaluated. The oedema development, measured at 180 min following carrageenan intraplantar injection, was significantly reduced by all tested doses: 75 mg/kg (33.30 percent), 150 mg/kg (45.80 percent) and 300 mg/kg (75.00 percent). The LD50 value was greater than 2000 mg/kg. These results demonstrated that the ethanol extract from Z. montana leaf possesses anti-nociceptive and anti-inflammatory activities, which could be of relevance for the pharmacological control of pain and inflammatory processes.

Anti-inflammatory and anti-nociceptive effects of Zeyheria montana (Bignoniaceae) ethanol extract.

In this work, the analgesic and anti-inflammatory activities of Zeyheria montana Mart. ethanol leaf extract were investigated at doses of 75, 150 and 300 mg/kg body weight. In the analgesic assay, against a chemical stimulus in mice, acetic acid-induced writhes were significantly inhibited by the extract at doses of 75 mg/kg (67.27%), 150 mg/kg (49.38%) and 300 mg/kg (82.87%). Also, a vigorous decrease in hyperalgesia was observed when measured after 2 h and 6 h of lipopolysaccharide stimulation of rats for all doses of extract tested. Z. montana extract, at doses of 75 and 300 mg/kg, caused very slight central analgesia in rats submitted to thermal stimulus, particularly noticeable at 30 min following treatment. The anti-inflammatory activity of Z. montana extract on carrageenan-induced oedema in rats was evaluated. The oedema development, measured at 180 min following carrageenan intraplantar injection, was significantly reduced by all tested doses: 75 mg/kg (33.30%), 150 mg/kg (45.80%) and 300 mg/kg (75.00%). The LD50 value was greater than 2000 mg/kg. These results demonstrated that the ethanol extract from Z. montana leaf possesses anti-nociceptive and anti-inflammatory activities, which could be of relevance for the pharmacological control of pain and inflammatory processes.

In vitro evaluation of the cytotoxic and trypanocidal activities of Ampelozizyphus amazonicus (Rhamnaceae)

Ampelozizyphus amazonicus Ducke is a tree commonly found in the Amazon region and an extract of its stem bark is popularly used as an antimalarial and anti-inflammatory agent and as an antidote to snake venom. Ursolic acid; five lupane type triterpenes: betulin, betulinic acid, lupenone, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid, and 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid, and three phytosteroids: stigmasterol, sitosterol and campesterol, have been isolated from stem extracts of A. amazonicus Ducke. Their structures were characterized by spectral data including COSY and HMQC. In an in vitro biological screening of the isolated compounds, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid was cytotoxic against the SKBR-3 human adenocarcinoma cell line (1 to 10 mg/mL), while 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid exhibited cytotoxicity against both SKBR-3 human adenocarcinoma and C-8161 human melanoma tumor cell lines (>0.1 mg/mL). In the present study, different extracts and some fractions of this plant were also investigated for trypanocidal activity due to the presence of pentacyclic triterpenes. The triterpene classes are potent against Trypanosoma cruzi. The bioassays were carried out using blood collected from Swiss albino mice by cardiac puncture during the parasitemic peak (7th day) after infection with the Y strain of T. cruzi. The results obtained showed that A. amazonicus is a potential source of bioactive compounds since its extracts and fractions isolated from it exhibited in vitro parasite lysis against trypomastigote forms of T. cruzi at concentrations >100 æg/mL. Fractions containing mainly betulin, lupenone, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid, and 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid showed more activity than crude extracts.

In vitro evaluation of the cytotoxic and trypanocidal activities of Ampelozizyphus amazonicus (Rhamnaceae).

Ampelozizyphus amazonicus Ducke is a tree commonly found in the Amazon region and an extract of its stem bark is popularly used as an antimalarial and anti-inflammatory agent and as an antidote to snake venom. Ursolic acid; five lupane type triterpenes: betulin, betulinic acid, lupenone, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid, and 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid, and three phytosteroids: stigmasterol, sitosterol and campesterol, have been isolated from stem extracts of A. amazonicus Ducke. Their structures were characterized by spectral data including COSY and HMQC. In an in vitro biological screening of the isolated compounds, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid was cytotoxic against the SKBR-3 human adenocarcinoma cell line (1 to 10 mg/mL), while 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid exhibited cytotoxicity against both SKBR-3 human adenocarcinoma and C-8161 human melanoma tumor cell lines (>0.1 mg/mL). In the present study, different extracts and some fractions of this plant were also investigated for trypanocidal activity due to the presence of pentacyclic triterpenes. The triterpene classes are potent against Trypanosoma cruzi. The bioassays were carried out using blood collected from Swiss albino mice by cardiac puncture during the parasitemic peak (7th day) after infection with the Y strain of T. cruzi. The results obtained showed that A. amazonicus is a potential source of bioactive compounds since its extracts and fractions isolated from it exhibited in vitro parasite lysis against trypomastigote forms of T. cruzi at concentrations >100 microg/mL. Fractions containing mainly betulin, lupenone, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid, and 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid showed more activity than crude extracts.

A 3' cis-regulatory region controls wingless expression in the Drosophila eye and leg primordia.

The precise regulation of wingless (wg) expression in the Drosophila eye disc is key to control the anteroposterior and dorsoventral patterning of this disc. Here, we identify an eye disc-specific wg cis-regulatory element that functions as a regulatory rheostat. Pannier (Pnr), a transcription factor previously proposed to act as an upstream activator of wg, is sufficient to activate the eye disc enhancer but required for wg expression only in the peripodial epithelium of the disc. We propose that this regulation of wg by Pnr appeared associated to the development of the peripodial epithelium in higher dipterans and was added to an existing mechanism regulating the deployment of wingless in the dorsal region of the eye primordium. In addition, our analysis identifies a separate ventral disc enhancer that lies adjacent to the eye-specific one, and thus altogether, they define a 1-kb genomic region where disc-specific enhancers of the wg gene are located.

Dynamics and function of intron sequences of the wingless gene during the evolution of the Drosophila genus.

To understand the function and evolution of genes with complex patterns of expression, such as the Drosophila wingless gene, it is essential to know how their transcription is regulated. However, extracting the relevant regulatory information from a genome is still a complex task. We used a combination of comparative genomics and functional approaches to identify putative regulatory sequences in two introns (1 and 3) of the wingless gene and to infer their evolution. Comparison of the sequences obtained from several Drosophila species revealed colinear and well-conserved sequence blocks in both introns. Drosophila willistoni showed a rate of evolution, in both introns, faster than expected from its phylogenetic position. Intron 3 appeared to be composed of two separate modules, one of them lost in the willistoni group. We tested whether sequence conservation in noncoding regions is a reliable indicator of regulatory function and, if this function is conserved, by analyzing D. melanogaster transgenic reporter lines harboring intron 3 sequences from D. melanogaster (Sophophora subgenus) and the species from the Drosophila subgenus presenting the most divergent sequence, D. americana. The analysis indicated that intron 3 contains pupal enhancers conserved during the evolution of the genus, despite the fact that only 30% of the D. melanogaster intron 3 sequences lie in conserved blocks. Additional analysis of D. melanogaster transgenic reporter lines harboring intron 3 sequences from D. willistoni revealed the absence of an abdomen-specific expression pattern, probably due to the above-mentioned loss of a regulatory module in this species.

Estudo comparativo da produção de metabólitos secundários em cultura de células e na planta in natura de Gomphrena globosa (Amaranthaceae)

A cultura de calos de Gomphrena globosa a partir de segmentos foliares foi estabelecida em meio contendo combinações dos fitoreguladores auxina/citocinina. Metabólitos secundários identificados através da fitoquímica da planta in natura, foram utilizados como padrões em análises por cromatografia líquida de alta eficiência e cromatografia de gás de extratos da cultura. O estudo comparativo revelou que os calos foram capazes de produzir alguns metabólitos sintetizados pela planta in natura.

The culture of calluses of Gomphrena globosa starting from segments foliate was established in half containing combinations of the phytoregulators auxina/citocinina. Identified secondary metabolites through the phytochemistry of the plant in natura were used as patterns in analyses for high performance liquid cromatography and gas cromatography of extracts of the culture. The comparative study revealed that the calluses were capable to produce some metabolite synthesized by the plant in natura.

The RGS domain-containing fission yeast protein, Rgs1p, regulates pheromone signalling and is required for mating.

BACKGROUND: When nutritionally starved, the fission yeast Schizosaccharomyces pombe enters a cell differentiation process which leads to mating and meiosis. The Ste11 protein is a key regulator of this differentiation pathway, activating the transcription of mating and meiotic genes upon starvation. RESULTS: Here, we describe rgs1, a member of the Regulator of G-protein Signalling (RGS) family, as a novel Ste11 target gene. rgs1 expression requires both an Ste11-mediated nitrogen starvation signal and the pheromone-induced activation of the Byr2/Byr1/Spk1 MAPK pathway. We show that rgs1 deletion results in a sensitivity to pheromone and in a mating defect. Deltargs1 cells initiate the mating pathway normally, undergoing sexual agglutination and G1 arrest, while inducing pheromone-dependent transcription, but then fail to fuse with a mating partner while elongating abnormal conjugation tubes. Endogenous Rgs1 tagged with GFP localizes to the nucleus and cytoplasm, and this localization pattern is not altered during pheromone treatment. Importantly, Rgs1 function requires its C-terminal RGS domain, as well as a central DEP domain and a novel homology domain present in its N-terminal region (Fungal-DR domain). CONCLUSIONS: These results demonstrate that rgs1 expression requires nutritional starvation and pheromone signalling. Rgs1 negatively regulates pheromone signalling during mating, acting in a negative feedback loop that is essential for the mating process.

Inhibition of the lethal and myotoxic activities of Crotalus durissus terrificus venom by Tabernaemontana catharinensis: identification of one of the active components.

In Brazilian folk medicine, victims of bites by poisonous animals are usually treated with plant extracts derived from the diverse national flora. The chemical and pharmacological properties of most extracts were yet not investigated. In the rural community of Assis-SP, the root bark of Tabernaemontana catharinensis ("leiteiro", "cow milk") is applied to the site of the snake bite and believed to neutralize the effect of the venom. We report here the ability of the lyophilized aqueous extract (AE) and of a pure compound obtained from the ethanolic extract of T. catharinensis to inhibit the lethal and myotoxic activities of C. d. terrificus (South American rattlesnake) venom. Doses of 10 mg AE/100 g, injected (i.m., rat) 20 s after injecting (i.m.) the venom and that of 2.5 mg AE/100 g, incubated for 1 h at 25 degrees C with the venom before injection (i.m.) were able to neutralize the lethal activity of 2LD50. These data indicate that T. catharinensis could be used as a source of a model molecule able to neutralize the lethality and myotoxicity induced by C. d. terrificus venom. Its ethanolic extract was then fractionated on a silica gel 60 chromatography column affording fractions A to F. Fraction A consisted basically of non-polar compounds, terpenes and sterols. Fraction D showed a pronounced antiophidian activity which was later correlated with the presence of the quaternary alkaloid 12-methoxy-4-methylvoachalotine in this fraction. This alkaloid was isolated and inhibited 100% lethality when injected 20 s after 2 LD50 at 1.7 mg/100 g.