Influência da luz LED na cultura celular de Mieloma múltiplo murino–NS1 in vitro

A monoclonal antibody (mAb) is an important tool in medical biotechnology and the production of biopharmaceuticals, especially for disease diagnosis and treatment of infections, because the antibodies have a significant advantage over chemical agents used in conventional therapies . The last thirty years the technology of production of monoclonal antibodies developed mainly the technique of obtaining in vitro, but also of their production is laborious, the cost is high. A major element of the high cost of production is the fact that the long-term culture consumes a large amount of imported inputs with high added value. A major contribution of this work is to promote cell growth more quickly and efficiently. Currently, a great race to discover new technologies and techniques to synthesize new antibodies and significantly increase the production of murine mAbs. New technologies such as laser and LED are innovations and widespread in modern life, so much so that its use has proliferated worldwide, primarily in the medical field. Recent studies show a series of results from the influence of the LED light in biological tissues such as: increasing the rate of cell proliferation, increased production rate of fibroblasts, increasing the rate of synthesis of RNA and DNA synthesis of ATP, etc. To assess the contribution of the LED in the culture of Myeloma NS1murino compared to the standard procedure. - NS1 cells were provided and followed the criteria of culture medium of the Laboratory of Cellular Engineering Center of Botucatu (POPs). The same amount of cells was grown in bottles of 25 cm2 polystyrene Tissue Culture Treated, specifically marked and kept in special medium RPMI 1640 Gibco BRL  supplemented with fetal bovine serum 10%, essential amino acids and non-essential, glucose, insulin and antibiotics. It was used in LEDs Cromatek wavelength of 630nm, 475nm and 530nm. The groups were... (Complete abstract click electronic access below)

Manejo da adubação fosfatada da aveia preta e do consórcio milho/capim Marandu com inoculação por Azospirillum brasilense em sistema plantio direto

Pós-graduação em Agronomia - FEIS

Desenvolvimento do cultivo 3D a partir de células primarias de neoplasias mamárias caninas: estudo da apoptose sobre efeito ou não da carboplatina

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

Evaluation of the genotoxicity of waters impacted by domestic and industrial effluents of a highly industrialized region of So Paulo State, Brazil, by the comet assay in HTC cells

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Benzilaminopurina e ácido naftaleno acético na indução e multiplicação in vitro de gemas de abacaxizeiro da cultivar 'IAC Gomo-de-mel'

O objetivo deste trabalho foi avaliar os efeitos de BAP (6-benzilaminopurina) e NAA (ácido naftaleno acético) na indução, na multiplicação in vitro de gemas, nas brotações de Ananas comosus da cultivar 'IAC Gomo-de-mel' e a correlação desses efeitos com a atividade de peroxidase e o teor de proteína solúvel total. Foram utilizadas gemas axilares retiradas da coroa de frutos sadios, inoculadas em tubo de ensaio contendo meio de cultura MS solidificado com ágar a 5%, pH ajustado para 5,7, contendo os tratamentos que incluíam diferentes concentrações e combinações de BAP (0, 0,5, 1,0 e 1,5mg L-1) e NAA (0, 0,5 e 1,0mg L-1). Nessa fase, aos 65 dias, ocorreu a formação de 2,24 brotações, utilizando-se 1mg L-1 de BAP. Após o desenvolvimento, as gemas foram inoculadas em meio MS líquido associado a dois tratamentos (1,0mg L-1 BAP + 0,5mg L-1 NAA e 1,0mg L-1 BAP + 1,0mg L-1 NAA) e, aos 95 dias, o meio de cultura mais adequado foi aquele que continha 1,0mg L-1 BAP + 0,5mg L-1 NAA, proporcionando 7,42 brotações, menor porcentagem de hiper-hidricidade, maior número de brotações e indução de gemas. As proteínas solúveis apresentaram relação negativa com hiper-hidricidade e comprimento de brotações. A atividade da peroxidase foi maior em plantas com maior número de brotos e com maior porcentagem de hiper-hidricidade.

Avaliação da aplicação exógena de poliaminas no crescimento de calos de mangabeira (Hancornia speciosa Gomes)

Este trabalho teve como objetivo estudar o efeito das poliaminas espermidina e espermina no crescimento de calos Hancornia speciosa Gomes. Calos com 0,5 cm de diâmetro foram inoculados em meio Murashige & Skoog (1962) (MS) a 50% + 100 mg L-1 de caseína hidrolisada + 200 mg L-1 de levedura de cerveja, variando os tratamentos:A: 1 mmol de espermina + 2 mg L-1 de 2,4-D (ácido 2,4 diclorofenoxiacético) + 0,5 mg L-1 de NAA (ácido naftalenoacético); B: 1 mmol de espermidina + 2 mg L-1 de 2,4-D + 0,5 mg L-1 de NAA; C: 2 mg L-1 de 2,4-D + 0,5 mg L-1 de NAA. Não houve influência das poliaminas no crescimento dos calos. observou-se, nos calos tratados com espermidina, maior concentração celular de putrescina (582,37 µg g mf-1) aos 60 dias, maior teor de espermidina (502,54 µg g mf-1) e espermina (868,53 µg g mf-1) aos 40 dias de cultivo, quando se aplicou a própria poliamina. Conclui-se que a aplicação exógena de poliaminas em Hancornia speciosa não proporciona aumento no crescimento de calos. A oxidação promovida por longos períodos de cultivo in vitro induz aumento nos níveis de putrescina.

Oxidation of amino acids by chlorpromazine cation radical and co-catalysis by chlorpromazine

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

Efeito do fator de crescimento epidermal (EGF) na maturação in vitro de oócitos caninos

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

Germinação de sementes de maracujazeiros: água, luz, temperatura e reguladores vegetais

The production of seedlings of passion fruit is done mainly with the use of seeds by most producers. It can also be propagated by cuttings, layering and in vitro tissue culture. However, even when grafting is used, it is necessary to use seeds to produce the rootstock. Nevertheless, most Passifloracea species have germination problems, making it difficult to obtain seedlings. Although several studies have been developed in an attempt to increase seed germination of passion fruit there are still difficulties in understanding the germination process and also disagreement about seed dormancy. Thus, the present study aimed to review papers related to seed germination of passion fruit emphasizing aspects such as mechanical resistance to water ingress, the need for adequate light and temperature and the use of plant growth regulators in overcoming seed dormancy. It was noticed that, in general, the passion fruit seeds are photoblastic negative and exhibit physiological dormancy, being no need of seed scarification to stimulate its germination.

Estudo da diversidade genética e química e produção do epímero do ácido caurenóico em calos de croton antisyphiliticus mart

Pós-graduação em Agronomia (Horticultura) - FCA

Estudo da diversidade genética e química e produção do epímero do ácido caurenóico em calos de croton antisyphiliticus mart

Pós-graduação em Agronomia (Horticultura) - FCA

Human immature dental pulp stem cells share key characteristic features with limbal stem cells

Objectives: Limbal stem cells (LSC) are self-renewing, highly proliferative cells in vitro, which express a set of specific markers and in vivo have the capacity to reconstruct the entire corneal epithelium in cases of ocular surface injury. Currently, LSC transplantation is a commonly used procedure in patients with either uni- or bilateral total limbal stem cells deficiency (TLSCD). Although LSC transplantation holds great promise for patients, several problems need to be overcome. In order to find an alternative source of cells that can partially substitute LSC in cornea epithelium reconstruction, we aimed at investigating whether human immature dental pulp stem cells (hIDPSC) would present similar key characteristics as LSC and whether they could be used for corneal surface reconstruction in a rabbit TLSCD model. Materials: We used hIDPSC, which co-express mesenchymal and embryonic stem cell markers and present the capacity to differentiate into derivative cells of the three germinal layers. TLSCD was induced by chemical burn in one eye of rabbits. After 30 days, the opaque tissue formed was removed by superficial keratectomy. Experimental group received undifferentiated hIDPSC, while control group only received amniotic membrane (AM). Both groups were sacrificed after 3 months. Results and conclusions: We have demonstrated, using immunohistochemistry and reverse transcription-polymerase chain reaction, that hIDPSCs express markers in common with LSC, such as ABCG2, integrin beta 1, vimentin, p63, connexin 43 and cytokeratins 3/12. They were also capable of reconstructing the eye surface after induction of unilateral TLSCD in rabbits, as shown by morphological and immunohistochemical analysis using human-specific antibodies against limbal and corneal epithelium. Our data suggest that hIDPSCs share similar characteristics with LSC and might be used as a potential alternative source of cells for corneal reconstruction.

Expression of soluble and functional full-length human matrix metalloproteinase-2 in Escherichia coli

Characterization of the matrix metalloproteinase-2 (MMP-2) substrates and understanding of its function remain difficult because up to date preparations containing minor amounts of other eukaryotic proteins that are co-purified with MMP-2 are still used. In this work, the expression of a soluble and functional full-length recombinant human MMP-2 (rhMMP-2) in the cytoplasm of Escherichia coli is reported, and the purification of this metalloproteinase is described. Culture of this bacterium at 18 degrees C culminated in maintenance of the soluble and functional rhMMP-2 in the soluble fraction of the E. coli lysate and its purification by affinity with gelatin-sepharose yielded approximately 0.12 mg/L of medium. Western Blotting and zymographic analysis revealed that the most abundant form was the 72-kDa MMP-2, but some gelatinolytic bands corresponding to proteins with lower molecular weight were also detected. The obtained rhMMP-2 was demonstrated to be functional in a gelatinolytic fluorimetric assay, suggesting that the purified rhMMP-2 was correctly folded. The method described here involves fewer steps, is less expensive, and is less prone to contamination with other proteinases and MMP inhibitors as compared to expression of rhMMP-2 in eukaryotic tissue culture. This protocol will facilitate the use of the full-length rhMMP-2 expressed in bacteria and will certainly help researchers to acquire new knowledge about the substrates and biological activities of this important proteinase. (C) 2011 Elsevier B.V. All rights reserved.

Genetic transformation of three sweet orange cultivars from explants of adult plants

The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of 'Hamlin', 'Pra', and 'Valencia' sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for 'Hamlin', 'Pra', and 'Valencia' sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L-1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.

IN VITRO ORGANOGENESIS OF RANGPUR LIME

Rangpur lime (Citrus limonia Osbeck) in vitro organogenesis was studied based on explant type and cytokinin culture media supplementation. Four explants types collected from epicotyl or hypocotyl regions of in vitro germinated seedlings were evaluated. The epicotyl-derived explants consisted of epicotyl segments and the hypocotyl-derived explants consisted of the entire hypocotyl segment, the hypocotyl segment attached to a cotyledon fragment, and the hypocotyl segment divided longitudinally. The explants were cultured on EME culture medium supplemented with benzylaminopurine (0, 0.5, 1.0, or 1.5 mg L-1). The evaluation was performed after 6 weeks. Best results considering both the explant responsiveness and number of shoots developed per explants were obtained when epicotyl segments-derived explants were evaluated. Considering the explant responsiveness of hypocotyl segments-derived explants no difference was detected between the entire hypocotyl segment and the hypocotyl segment attached to a cotyledon fragment. Moreover, the percentage of responsive explants decreased when hypocotyl segments divided longitudinally were tested. No difference was detected for the number of shoots developed per explant considering the three types of hypocotyl-derived explants. Culture media supplementation with BAP was not essential for Rangpur lime in vitro organogenesis. However, adventitious shoot development was stimulated in concentrations between 0.5 - 1.0 mg L-1.