Silenciamento da expressão da enzima óxido nítrico sintase neuronal (nNOS) em linhagem de neuroblastoma via siRNAs sintéticos em dupla fita

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, Programa de Pós-Graduação em Saúde Animal, 2011.

Especificidad del hongo micorrizico (Rhizoctonia sp.) en Phalaenopsis sp., Cymbidium sp., Trichoceros antenifer, Oncidium excavatum, y Cyrtochilum sp.

Las orquídeas producen abundantes semillas pequeñas, careciendo de endospermo, cotiledones y sustancias de reserva para llevar a cabo su germinación. Es por esto que estratégicamente las semillas establecen una relación simbiótica con un hongo micorrízico que favorece a su germinación y desarrollo. El objetivo de este estudio fue determinar la especificidad del hongo micorrízico (Rhizoctonia sp.) en la germinación de cinco géneros de orquídeas. Se usaron dos medios de cultivo: 1) PhytamaxTM y 2) avena-agar+Rhizoctonia sp. Los resultados evaluados a los 45 y 75 días demuestran que no existe especificidad entre el hongo Rhizoctonia sp. y Trichoceros antenifer, especie de la cual se aisló el hongo. La germinación en tres de los cinco géneros evaluados fue mayor en el tratamiento avena-agar+Rhizoctonia sp., evidenciando también mayor tamaño y vigor las plantas que se desarrollaron en este tratamiento, aunque estos datos no fueron evaluados. Los resultados sugieren también que la planta hospedera del hongo inoculado podría tener una ventaja en cuanto al tiempo requerido para su germinación; sin embargo otras, especies también se ven favorecidas con el inóculo, aunque éste sea obtenido de otra fuente.

Cultivo de Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) em laboratório utilizando meio CHU12 e de macrófita com NPK

Two different culture media, namely CHU12 and inorganic fertilizer NPK (20-5-20) associated with macrophyte (Eichhornia crassipes) extract, were used to evaluate the development of A. gracilis. Growth rate, development, nutritional value and medium water quality were analyzed. A. gracilis in macrophyte+NPK medium had mean cell density (333x10(5) cells/mL) higher than medium CHU12 (302x10(5) cells/mL). Protein rate (12.52% PS), dry weight (397x10(7) pg/cell) and ashes (4.08x10(7) pg/cell) in A. gracilis was higher (p<0.05) than medium macrophyte+NPK. on the other hand, lipids, carbohydrates and fibers had the same rate (p>0.05) in both media. When hydrological variables in the culture medium of A. gracilis are taken into account, dissolved oxygen and free CO2 alone failed to have any significant difference (p>0.05) between the media employed. A. gracilis in the macrophyte+NPK medium had the same nutritional rate as the commercial medium CHU12, with excellent results for alga growth. Macrophyte medium may be used for large scale alga culture in the feed of fish larvae.

Use of Bioscreen C for growth of Mucor hiemalis in different carbon and nitrogen sources

O sistema automatizado Bioscreen C foi utilizado para o crescimento de quatro linhagens de Mucor hiemalis, isoladas do solo da Estação Ecológica de Juréia-Itatins (EEJI), estado de São Paulo, em meios líquidos com uma única fonte de carbono (2%) ou de nitrogênio (1%), pH 5,0, a 25ºC, e agitação de 150rpm por 5 dias. O meio com somente uma única fonte de nitrogênio foi adicionado com 2% de glicose. As leituras de densidade óptica foram realizadas a 540nm, em intervalos de 2h, por cinco dias. Os resultados foram analisados estatisticamente com o Teste de Friedman (alfa = 5%). Os melhores crescimentos foram obtidos com as linhagens M1, M2 e M3, que atingiram o início da fase log em 60 horas de cultivo. As melhores fontes de carbono variaram de acordo com a linhagem estudada, e extrato de levedura provou ser a melhor fonte de nitrogênio para todas as linhagens. Acetato de sódio inibiu o crescimento das quatro linhagens, sendo a M3 a mais afetada. O uso do sistema automatizado foi muito conveniente para as culturas em meio liquido, sendo rápido e automático, constituindo em uma boa técnica para a determinação das condições ambientais ótimas para crescimento de fungos filamentosos.

Germinação de sementes e otimização de técnicas de micropropagação de umbuzeiro (spondias tuberosa, arr.) anacardiaceae

The brazilian-plum (Spondias tuberosa, His) is a tropical fruit tree that has been consolidated in the market for agribusiness processing, due to its characteristic flavor of fruit. Accordingly, studies to optimize the propagation of plants are necessary for production of seedlings with agronomic and quality assurance measures. This study aimed at determining the efficient techniques for uniform seed germination, as brazilian-plum seed present mechanical dormancy, and establish optimal culture media for multiplication of shoots from the in vitro micropropagation. Firstly, in a greenhouse at the Universidade Federal do Rio Grande do Norte, was evaluated the influence of different methods of breaking dormancy in the emergence of seedlings of brazilian-plum and speed of germination (IVG) of seeds. After 60 days of cultivation, it was found that splay in the distal portion of the seed was the best treatment, with rates of 85.33% in germinability and 3.415 of IVG, compared with the treatment of seed-soaking in water for 12h + humus and the control group. Subsequently, new sources of seedling explants were obtained in studies of tissue culture. Laboratory of Plant Biotechnology that the university, was used stem apex, nodal segments and internodes in search of decontamination with various concentrations of calcium hypochlorite [Ca(OCl)2] and micropropagation, inoculating them in half WPM (1980) with various concentrations of 6-benzylaminopurine (BAP). We used 10 sample units with three replications for different concentrations of [Ca(OCl)2], BAP and explants type. After thirty days, which was observed for the control of contamination, during the establishment in vitro, concentrations of [Ca(OCl)2] between 0.5% and 2.0% were effective in combating exogenous contamination of the apex. In nodal segments and internodes, concentrations of [Ca(OCl)2] between 1.0% and 2.0% and 1.5% and 2.0% were respectively, sufficient to reduce the percentage of losses in these infestations explants. For micropropagation, the culture medium supplemented with 0.1 mg.L-1 BAP promotes better development of multiple shoots per explants from nodal segment. However, success does not get to shoot training in internodal segment

Meios de transporte de dentes avulsionados

A avulsão dentária é um dos mais severos traumatismos dento-alveolares, caracterizado pela desinserção completa do dente do seu ambiente natural, o alvéolo. O prognóstico desta condição traumática depende em grande escala da atitude adoptada no momento do acidente. No entanto, quando o reposicionamento dentário imediato não é possível, factores como o tempo que o dente permaneceu no meio extra-oral e o meio de transporte utilizado influenciam consideravelmente o prognóstico da peça dentária avulsionada. A elaboração deste trabalho tem como objectivo verificar o estado actual da investigação científica relativamente aos meios de transporte de dentes avulsionados, nomeadamente, indagar sobre os potenciais meios de transporte actualmente em estudo. Foi efectuada uma pesquisa bibliográfica digital, na base de dados PUBMED, com os seguintes termos de pesquisa: “tooth avulsion”, “storage media”, “tooth reimplantation” e “delayed reimplantation” isoladamente ou em combinação. Para a selecção dos artigos foram estipulados critérios de inclusão e exclusão na pesquisa bibliográfica. Existem soluções especificamente concebidas para a conservação e preservação de células humanas, que idealmente deveriam ser sempre utilizadas como meios de transporte de dentes avulsionados, tais como: solução balanceada de Hank, Viaspan®, Eurocollins® e determinados meios de cultura. São soluções que apresentam propriedades biofísico-químicas que os levam a ser considerados meios de transporte de eleição. No entanto, a sua escassa disponibilidade no momento da avulsão desperta a necessidade pela investigação de outros meios, nomeadamente, soluções que apresentem nesse contexto maior disponibilidade. O leite, a saliva, a solução salina e o Gatorade® continuam a representar meios alternativos para o transporte de dentes avulsionados, porém, outras soluções emergentes apresentam características biológicas que as remetem para meios de transporte promissores como por exemplo: solução de propólis, o chá verde, o extracto de aloe Vera, água de côco, clara de ovo, extracto de amora, sumo de romã, salvia officinalis e Ricetral®. Evidencia-se, com este trabalho, a necessidade de estudos futuros nesta área de modo a que meios de transporte que actualmente pensa-se poderem ser extremamente efectivos na manutenção da viabilidade célular e com ampla disponibilidade, possam ser utilizados de forma segura e vantajosa para o paciente, nomeadamente, garantindo melhores prognósticos em situações de reimplantação dentária.

In vitro Culture of Bovine Embryos in Murine ES Cell Conditioned Media Negatively Affects Expression of Pluripotency-Related Markers OCT4, SOX2 and SSEA1

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

Establishing a high titer transient gene expression process in conditioned media for CHO-DG44 cells

Dissertação para obtenção do Grau de Mestre em Biotecnologia

A proteome analysis of conditioned media from human neonatal fibroblasts used in the maintenance of human embryonic stem cells

The pathways involved in the maintenance of human embryonic stem (hES) cells remain largely unknown, although some signaling pathways have been identified in mouse embryonic stem (mES) cells. Fibroblast feeder layers are used to maintain the undifferentiated growth of hES cells and an examination of the conditioned media (CM) of human neonatal fibroblasts (HNFs) could provide insights into the maintenance of hES cells. The neonatal foreskin fibroblast line (HNF02) used in this study was shown to have a normal 2n = 46, XY chromosomal complement and to support the undifferentiated growth of the Embryonic Stem Cell International Pte. Ltd.-hES3 cell line. The CM of HNF02 was examined using two-dimensional liquid chromatography-tandem mass spectrometry (2-D LCMS) and two-dimensional electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry (2-DE/MALDI). A total of 102 proteins were identified, 19 by 2-DE/MALDI, 53 by 2-D LCMS and 30 by both techniques. These proteins were classified into 15 functional groups. Proteins identified in the extracellular matrix and differentiation and growth factor functional categories were considered most likely to be involved in the maintenance of hES cell growth, differentiation and pluripotency as these groups contained proteins involved in a variety of events including cell adhesion, cell proliferation and inhibition of cell proliferation, Writ signaling and inhibition of bone morphogenetic proteins.

Ex-ante evaluation by the media of the Guimarães 2012 European Capital of Culture

The nomination of Guimarães to host the 2012 European Capital of Culture (ECC) has put on the agenda of the city the need of measuring the effects that the implementation of this mega event could have in it and in the municipality a whole. The balance of the benefits and costs and an extended community involvement tend to reduce negative impacts and enhance positive ones. This chapter analyzes the involvement of population and local associations in the planning and organization of the 2012 Guimarães European Capital of Culture, using the coverage made during 2011 by local and national press of the mega event. A content analysis of the news published covering the period between January and December 2011 and using three newspapers was conducted. From those, two were local and weekly newspapers and one was a national daily one. Looking to data results, it can be concluded that it was poor the community involvement and, also, the one of the cultural associations in the organizations of the 2012 ECC. A strong negative reaction to the model choose to plan the mega event conducted by official organizers was found, which has cast doubts on the desirable participation of the residents and, consequently, on the success of the mega event, especially in a perspective of a medium and long term effects.

Ex-ante evaluation by the media of the Guimarães 2012 European Capital of Culture

The nomination of Guimarães to host the 2012 European Capital of Culture (ECC) has put on the agenda of the city the need of measuring the effects that the implementation of this mega event could have in it and in the municipality a whole. The balance of the benefits and costs and an extended community involvement tend to reduce negative impacts and enhance positive ones. This chapter analyzes the involvement of population and local associations in the planning and organization of the 2012 Guimarães European Capital of Culture, using the coverage made during 2011 by local and national press of the mega event. A content analysis of the news published covering the period between January and December 2011 and using three newspapers was conducted. From those, two were local and weekly newspapers and one was a national daily one. Looking to data results, it can be concluded that it was poor the community involvement and, also, the one of the cultural associations in the organizations of the 2012 ECC. A strong negative reaction to the model choose to plan the mega event conducted by official organizers was found, which has cast doubts on the desirable participation of the residents and, consequently, on the success of the mega event, especially in a perspective of a medium and long term effects.

Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µ L and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter time. These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias) or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits.