Efeito acaricida in vitro de extratos de plantas do Pantanal no carrapato de bovinos, Rhipicephalus (Boophilus) microplus.

Estudou-se a ação acaricida dos extratos brutos etanólicos de 14 plantas do Pantanal sobre fêmeas ingurgitadas do carrapato-do-boi. Os bioensaios foram realizados em triplicatas pelo método de imersão das teleóginas e estimadas a produção de ovos, a eclodibilidade e a eficácia dos extratos. Extratos do lenho da raiz e da casca da raiz de Annona dioica, da raiz e da casca do caule de Simarouba versicolor, da raiz de Annona cornifolia e de Duguetia furfuracea tiveram atividade acaricida entre 50% e 100% e resposta dose-dependente. Extratos de Dimorphandra mollis, Magonia pubescens, Protium heptaphyllum, Hyptis crenata, Sebastiana hispida, Aspidosperma australe, Senna occidentalis e de Elyonurus muticus mostraram atividade acaricida baixa ou ausente (0% a 10%). Extratos da parte aérea de A. cornifolia, da planta inteira de Croton glandulosus e da casca do caule de Stryphnodendron obovatum mostraram atividade acaricida intermediária (10% a 20%). A alta atividade acaricida observada nos extratos de A. dioica, A. cornifolia e D. furfuracea indica ação de acetogeninas, moléculas naturais com várias atividades farmacológicas já descritas e presentes, principalmente, nas plantas da família Annonaceae.

Preservação de germoplasma vegetal, com ênfase na conservação in vitro de variedades de mandioca.

O melhoramento genético de plantas tem contribuído sobremaneira para o aumento da produção em diversas espécies de grande importância econômica e/ou social, beneficiando bilhões de pessoas, especialmente de menor poder aquisitivo, que vivem em países em desenvolvimento distribuídos por todo o mundo. Esses aumentos na produção resultam da obtenção de novos genótipos, que apresentam rendimentos mais elevados, adaptados a diversas condições ecológicas, muitas das vezes adversas, e resistentes a pragas e doenças. No entanto, para a geração desses materiais melhorados, torna-se necessário que características de interesse sejam incorporadas às novas cultivares, dentro de programas de melhoramento genético bem definidos, de forma que possam ser, ao final de todo o processo, exploradas comercialmente.

Extracao de DNA Spiroplasma kunkelii Whitcomb cultivado in vitro.

2003

Development of an in vitro multicellular tumor spheroid model using microencapsulation and its application in anticancer drug screening and testing

In this study, an in vitro multicellular tumor spheroid model was developed using microencapsulation, and the feasibility of using the microencapsulated. multicellular tumor spheroid (MMTS) to test the effect of chemotherapeutic drugs was investigated. Human MCF-7 breast cancer cells were encapsulated in alginate-poly-L-lysine-alginate (APA) microcapsules, and a single multicellular spheroid 150 mu m in diameter was formed in the microcapsule after 5 days of cultivation. The cell morphology, proliferation, and viability of the MMTS were characterized using phase contrast microscopy, BrdU-Iabeling, MTT stain, calcein AM/ED-2 stain, and H&E stain. It demonstrated that the MMTS was viable and that the proliferating cells were mainly localized to the periphery of the cell spheroid and the apoptotic cells were in the core. The MCF-7 MMTS was treated with mitomycin C (MC) at a concentration of 0.1, 1, or 10 times that of peak plasma concentration (ppc) for up to 72 h. The cytotoxicity was demonstrated. clearly by the reduction in cell spheroid size and the decrease in cell viability. The MMTS was further used to screen the anticancer effect of chemotherapeutic drugs, treated with MC, adriamycin (ADM) and 5-fluorouracil (5-FU) at concentrations of 0.1, 1, and 10 ppc for 24, 48, and 72 h. MCF-7 monolayer culture was used as control. Similar to monolayer culture, the cell viability of MMTS was reduced after treatment with anticancer drugs. However, the inhibition rate of cell viability in MMTS was much lower than that in monolayer culture. The MMTS was more resistant to anticancer drugs than monolayer culture. The inhibition rates of cell viability were 68.1%, 45.1%, and 46.8% in MMTS and 95.1%, 86.8%, and 91.6% in monolayer culture treated with MC, ADM, and 5-FU at 10 ppc for 72 h, respectively. MC showed the strongest cytotoxicity in both MMTS and monolayer, followed by 5-FU and ADM. It demonstrated that the MMTS has the potential to be a rapid and valid in vitro model to screen chemotherapeutic drugs with a feature to mimic in vivo three-dimensional (3-D) cell growth pattern.

Germinação in vitro do pólen de cupuaçuzeiro.

Estudo de vários parâmetros, como a caracterização de cinco estádios da antese da flor de cupuaçuzeiro e a determinação do tamanho da amostra para a contagem dos grãos de pólen germinados, com o objetivo de desenvolver uma metodologia para germinação in vitro do pólen de cupuaçuzeiro.

Protocolo para a propagação in vitro de bananeira cultivar prata zulu.

Este trabalho tem por objetivo relatar, de forma simplificada, a partir de resultados de pesquisas realizadas no Laboratório de Biotecnologia da Embrapa Amazônia Ocidental, Manaus, as melhores condições de produção in vitro de mudas da banana cv. Prata Zulu, cultivar que tem sobressaído às demais pela alta resistência à sigatoka-negra e pelos bons resultados fitotécnicos apresentados.

Adaptação de uma técnica "in vitro" para descrição da cinética de degradação ruminal da matéria seca de volumosos.

Este trabalho teve como objetivo adaptar a fase fermentativa da técnica de digestibilidade in vitro proposta por Tilley e Terry (1963) para a descrição da cinética de degradação ruminal da matéria seca (MS) de volumosos. Para tal, comparou os resultados da cinética de degradação ruminal obtidos pela técnica in situ aos resultados obtidos pela fase fermentativa da técnica de Tilley e Terry (1963) empregando diferentes tempos de incubação das amostras de forrageiras.

Efeito da proporcao NH4+/NO3- na distribuicao de nitrato e na atividade in vitro da redutase do nitrato de plantas de Panicum maximum.

O objetivo deste trabalho foi estudar os efeitos da nutricao nitrica e amoniacal na distribuicao de nitrato e na atividade in vitro da redutase do nitrato em plantas de capim coloniao ( Panicum maximum Jacq.). as plantas receberam 7 mM de nitrogenio nas forma N- NH4 +, N-NO3- ou combinadas nas seguintes proporcoes 0/100, 25/75, 50/50, 75/25 e 100/0. A extracao enzigmatica foi feita em Tris-HCI 50mM pH 7,5, EDTA 0,5 mM e b-mercaptoetanol 10mM, e para o ensaio utilizou-se 200 mmoles de Tris-HCI pH 7,5, 20 mmoles KNO3, 0,1 mmol NADH e 0,1 mL de Triton X-100 1% no volume final de 2,0 mL. A concentracao de nitrato foi levemente superior na parte aerea quando as plantas receberam ambas as formas de nitrogenio, e na raiz seta decresceu com o aumento da proporcao de aminio. A atividade in vitro de redutase do nitrato so foi observada nas folhas e foi maior quando as plantas receberam ambas as formas de nitrogenio. Os resultados sugerem um efeito estimulador do amonio no tranporte de nitrato e na atividade de redutase do nitrato quando NO3- esta presente.

Relação entre pressão e volume para implantação da técnica in vitro semi-automática de produção de gases na Embrapa Cerrados.

2008

Biologia da lagarta-do-paricá Syssphinx molina (Cramer, 1780) (Lepidoptera: Saturniidae) em dietas artificiais e naturais in vitro.

2015

Comparative proteomics of excretory-secretory proteins released by the liver fluke Fasciola hepatica in sheep host bile and during in vitro culture ex host

Russell M. Morphew, Hazel A. Wright, E. James LaCourse, Debra J. Woods and Peter M. Brophy (2007). Comparative proteomics of excretory-secretory proteins released by the liver fluke Fasciola hepatica in sheep host bile and during in vitro culture ex host. Molecular and Cellular Proteomics, 6 (6), 963-972. Sponsorship: BBSRC / EU RAE2008

Cellular and in-vitro models to assess antioxidant activities of seaweed extracts and the potential use of the extracts as ingredients

Seaweeds contain a range of antioxidant compounds such as polyphenols, carotenoids, sulphated polysaccharides and vitamins and have the potential to be used as ingredients in neutraceuticals. The antioxidant activity of crude 60% methanol extracts prepared from five Irish seaweeds, Ascophyllum nodosum, Laminaria hyperborea, Pelvetia canaliculata, Fucus vesiculosus and Fucus serratus were examined using in-vitro assays and a cell model system to determine the antioxidant activity of the extracts and their ability to protect against H2O2 and tert-BOOH-induced DNA damage and alterations in cellular antioxidant status in the human adenocarcinoma, Caco-2 cell line. To optimise the extraction of antioxidant compounds from seaweeds, an accelerated solvent extraction (ASE®) was used in combination with food grade solvents. The antioxidant activity of these extracts against H2O2 and tert-BOOH-induced DNA damage and alterations in cellular antioxidant status was also assessed. Extracts that exhibited the highest antioxidant activity, A. nodosum (100% water and 80% ethanol extracts) and F. vesiculosus (60% ethanol extract) were selected as ingredients for incorporation into fluid milk and yogurt at concentrations of 0.25% and 0.5%. The addition of the seaweed extracts to milk and yogurt did not affect the pH or shelf-life properties of the products. Seaweed addition did however significantly influence the colour properties of the milk and yogurt. Yellowness values were significantly higher in yogurts containing F. vesiculosus at both concentrations and A. nodosum (80% ethanol) at the 0.5% concentration. In milk, the F. vesiculosus (60% ethanol) and A. nodosum (80% ethanol) at both the 0.25% and the 0.5% concentrations had higher greenness and yellowness values than the milk containing A. nodosum (100% water). Sensory analysis revealed that appearance and flavour governed the overall acceptability of yogurts with the control yogurt, and yogurts containing A. nodosum (100% water) were the most preferred samples by panellists. However, in the milk trial the perception of a fishy taste was the determining factor in the negative perception of milk. The unsupplemented control and the milk containing A. nodosum (100% water) at a concentration of 0.5% were the most overall accepted milk samples by the sensory panellists. The antioxidant activity of the extracts in milk and yogurt remained stable during storage as determined by the in-vitro assays. Seaweed supplemented milk and yogurt were also subjected to an in-vitro digestion procedure which mimics the human digestive system. The milk and yogurt samples and their digestates were added to Caco-2 cells to investigate their antioxidant potential however neither the undigested or digested samples protected against H2O2-induced DNA damage in Caco-2 cells.

Fundamental studies of sourdoughs fermented with Weissella cibaria and Lactobacillus plantarum: influence on baking characteristics, sensory profiles and in vitro starch digestibility of gluten-free breads

The application of sourdough can improve texture, structure, nutritional value, staling rate and shelf life of wheat and gluten-free breads. These quality improvements are associated with the formation of organic acids, exopolysaccharides (EPS), aroma or antifungal compounds. Initially, the suitability of two lactic acid bacteria strains to serve as sourdough starters for buckwheat, oat, quinoa, sorghum and flours was investigated. Wheat flour was chosen as a reference. The obligate heterofermentative lactic acid bacterium (LAB) Weissella cibaria MG1 (Wc) formed the EPS dextran (a α-1,6-glucan) from sucrose in situ with a molecular size of 106 to 107 kDa. EPS formation in all breads was analysed using size exclusion chromatography and highest amounts were formed in buckwheat (4 g/ kg) and quinoa sourdough (3 g/ kg). The facultative heterofermentative Lactobacillus plantarum FST1.7 (Lp) was identified as strong acidifier and was chosen due to its ubiquitous presence in gluten-free as well as wheat sourdoughs (Vogelmann et al. 2009). Both Wc and Lp, showed highest total titratable acids in buckwheat (16.8 ml; 26.0 ml), teff (16.2 ml; 24.5 ml) and quinoa sourdoughs (26.4 ml; 35.3 ml) correlating with higher amounts of fermentable sugars and higher buffering capacities. Sourdough incorporation reduced the crumb hardness after five days of storage in buckwheat (Wc -111%), teff (Wc -39%) and wheat (Wc -206%; Lp -118%) sourdough breads. The rate of staling (N/ day) was reduced in buckwheat (Ctrl 8 N; Wc 3 N; Lp 6 N), teff (Ctrl 13 N; Wc 9 N; Lp 10 N) and wheat (Ctrl 5 N; Wc 1 N; Lp 2 N) sourdough breads. Bread dough softening upon Wc and Lp sourdough incorporation accounted for increased crumb porosity in buckwheat (+10.4%; +4.7), teff (+8.1%; +8.3%) and wheat sourdough breads (+8.7%; +6.4%). Weissella cibaria MG1 sourdough improved the aroma quality of wheat bread but had no impact on aroma of gluten-free breads. Microbial shelf life however, was not prolonged in any of the breads regardless of the starter culture used. Due to the high prevalence of insulin-dependent diabetes mellitus particular amongst coeliac patients, glycaemic control is of great (Berti et al. 2004). The in vitro starch digestibility of gluten-free breads with and without sourdough addition was analysed to predict the GI (pGI). Sourdough can decrease starch hydrolysis in vitro, due to formation of resistant starch and organic acids. Predicted GI of gluten-free control breads were significantly lower than for the reference white wheat bread (GI=100). Starch granule size was investigated with scanning electron microscopy and was significantly smaller in quinoa flour (<2 μm). This resulted in higher enzymatic susceptibility and hence higher pGI for quinoa bread (95). Lowest hydrolysis indexes for sorghum and teff control breads (72 and 74, respectively) correlate with higher gelatinisation peak temperatures (69°C and 71°C, respectively). Levels of resistant starch were not increased by addition of Weissella cibaria MG1 (weak acidifier) or Lactobacillus plantarum FST1.7 (strong acidifier). The pGI was significantly decreased for both wheat sourdough breads (Wc 85; Lp 76). Lactic acid can promote starch interactions with gluten hence decreasing starch susceptibility (Östman et al. 2002). For most gluten-free breads, the pGI was increased upon sourdough addition. Only sorghum and teff Lp sourdough breads (69 and 68, respectively) had significantly decreased pGI. Results suggest that the increase of starch hydrolysis in gluten-free breads was related to mechanism other than presence of organic acids and formation of resistant starch.