21 resultados para Cell suspension

em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"


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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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This study aimed to assess antioxidant effects of melatonintreatment compared to N-acetylcysteine (NAC) and to their combination in asickle cell suspension. Sickle erythrocytes were suspended in phosphate-buffered saline, pH 7.4, composing external control group. They were alsosuspended and incubated at 37°C either in the absence (experimental controlgroup) or in the presence of NAC, melatonin and their combination atconcentrations of 100 pM, 100 nM and 100 lM for 1 hr (treatment groups).The melatonin influences were evaluated by spectrophotometric [hemolysisdegree, catalase (CAT), glutathione S-transferase (GST), glutathioneperoxidase (GPx), glutathione reductase (GR), glucose-6-phosphatedehydrogenase (G6PDH), and superoxide dismutase (SOD) activities] andchromatographic methods [glutathione (GSH) and malondialdehyde (MDA)levels]. Incubation period was able to cause a rise about 64% on hemolysisdegree as well as practically doubled the lipid peroxidation levels (P < 0.01).However, almost all antioxidants tested treatments neutralized this incubationeffect observed in MDA levels. Among the antioxidant biomarkers evaluated,we observed a modulating effect of combined treatment on GPx and SODactivities (P < 0.01), which showed ~25% decrease in their activities. Inaddition, we found an antioxidant dose-dependent effect for melatonin onlipid peroxidation (r = 0.29; P = 0.03) and for combined antioxidanttreatments also on MDA levels (r = 0.37; P = 0.01) and on SOD activity(r = 0.54; P < 0.01). Hence, these findings contribute with important insightthat melatonin individually or in combination with NAC may be useful forsickle cell anemia management.

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Objective To assess the expression of rabies virus G-glycoprotein (RVGP) expression using Semliki Forest virus as a vector in combination with BHK-21 cells cultured in suspension. Results A multilevel factorial design was used to quantify effects of temperature (33–37 C), fresh medium addition after the viral adsorption step (100–200 % with respect to the initial cell suspension volume before infection) and harvest time (8–40 h) on RVGP production. Experimental runs were performed in 24-well cell culture plates at a multiplicity of infection (MOI) of 16. An additional experiment in spinner-flask was performed at MOI of 9, using the optimal conditions determined in cell culture plates. Values for temperature, fresh medium addition and harvest time of 33 C, 100 % and 16 h, respectively, ensured the optimal RVGP production in culture plates. The volumetric yield (239 ng ml-1 ) in these conditions was higher than that reported previously for adherent cell culture. In spinner-flasks, the volumetric yield was improved (559 ng ml-1 ). Conclusion These results establish the basis for designing bioprocess to produce RVGP.

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The objective of this study was to evaluate the methodology to establish the hemolytic activity of alternative complement pathway as an indicator of the innate immunity in Brazilian fish pacu (Piaractus mesopotamicus), in addition to verifying the influence of β-glucan as an immunostimulant. Fish were fed with diets containing 0, 0.1 and 1% β-glucan, during seven days, and then inoculated with Aeromonas hydrophila. Seven days after the challenge, they were bled for serum extraction. The methodology consisted of a kinetic assay that allows calculating the required time for serum proteins of the complement to promote 50% lysis of a rabbit red blood cell suspension. The method developed in mammals was successfully applied for pacu and determined that the hemolytic activity of the proteins of the complement system (alternative pathway) increased after the pathogen challenge, but was not influenced by the β-glucan treatment.

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O presente trabalho objetivou gerar informações referentes à agressividade de linhagens de Xanthomonas axonopodis pv. aurantifolii Tipo C(Xaa-C), produtoras (PP) e não produtoras de pigmento (NP) escuro em meio de cultura, comparativamente a X. axonopodis pv. citri Tipo A (Xac) . Os tratamentos foram formados por 14 linhagens, sendo sete de Xaa-C PP, cinco Xaa-C NP, e duas linhagens de Xac. As linhagens foram inoculadas através de ferimentos, em folhas de lima ácida 'Galego' (Citrus aurantifolia Swingle), com agulha previamente mergulhada em uma suspensão de células bacterianas (10(7) UFC/mL). Foram realizadas dez repetições para cada tratamento, representadas por uma planta cada. As plantas foram mantidas em casa de vegetação durante todo o experimento. As linhagens diferiram entre si quanto ao período de incubação, diâmetro e populações bacterianas das lesões e, comparativamente, Xaa-C NP mostraram-se mais agressivas do que Xaa-C PP. Algumas linhagens induziram sintomas que diferiram quanto à presença e extensão de anasarca, halo amarelo e saliência do tecido necrosado.

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The aim of this study was to evaluate the effects of the laser radiation (685 nm) associated with photosensitizers on viability of different species of Candida genus. Suspensions of Candida albicans, Candida dubliniensis, Candida krusei and Candida tropicalis, containing 106 viable cells per milliliter were obtained with the aid of a Neubauer's chamber. From each species, 10 samples of the cell suspension were irradiated with diode laser (685 nm) with 28 J/cm(2) in the presence of methylene blue (0.1 mg/ml), 10 samples were only treated with methylene blue, 10 samples were irradiated with laser in the absence of the dye, 10 samples were treated with the dye and irradiated with laser light and 10 samples were exposed to neither the laser light nor to the methylene blue dye. From each sample, serial dilutions of 10(-2) and 10(-3) were obtained and aliquots of 0.1 ml of each dilution were plated in duplicate on Sabouraud dextrose agar. After incubation at 37 degrees C for 48 h, the number of colony-forming units (CFU/ml) was obtained and data were submitted to ANOVA and Tukey's test (p < 0.05). Laser radiation in the presence of methylene blue reduced the number of CFU/ml in 88.6% for C. albicans, 84.8% for C. dubliniensis, 91.6% for C krusei and 82.3% for C tropicalis. Despite of this, only laser radiation or methylene blue did not reduce significantly the number of CFU/ml of Candida samples, except for C tropicalis. It could be concluded that the photo activation of methylene blue by the red laser radiation at 685 nm presented fungicide effect on all Candida species studied. (c) 2006 Elsevier B.V. All rights reserved.

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Purpose: To analyze, in vitro, the effects of acetylsalicylic acid (aspirin) and acetic acid solutions on VX2 carcinoma cells in suspension and to examine the correlation between these effects and neoplastic cell death. Methods: The VX2 tumor cells (107 cells/ml) were incubated in solutions containing differing concentrations (2.5% and 5%) of either acetylsalicylic acid or acetic acid, or in saline solution (controls). Every five minutes, cell viability was tested (using the trypan blue test) and analyzed under light microscopy. Results: Tumor cell viability (in %) decreased progressively and, by 30 minutes, neoplastic cell death had occurred in all solutions. Conclusion: Based on this experimental model and the methodology employed, we conclude that these solutions cause neoplastic cell death in vitro.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Zymomonas mobilis was immobilized using a cell suspension fixed to 8.6 x 10(7) CFU mL(-1) by spectrophotometry. This biomass was suspended in sodium alginate solution (3%) that was dropped with a hypodermic syringe into 0.2 M calcium chloride solution. Was test two initial pH of fermentation medium (4 and 5) and different sucrose concentrations 15, 20, 25, 30 and 35% at 30 degrees C, without stirring for 24, 48, 72 and 96 hours. The levan production to pH 4 was high in sucrose 25% for 24 (16.51 g L-1) and 48 (15.31 g L-1) hours. The best values obtained to pH 5 was in sucrose 35% during 48 (22.39 g L-1) and 96 (23.5 g L-1) hours, respectively. The maximum levan yield was 40.8% and 22.47% in sucrose 15% to pH 4 and 5, respectively. Substrate consumption to pH 4 was bigger in sucrose 15 (56.4%) and 20% (59.4%) and to pH 5 was in 25 (68.85%) and 35% (64.64%). In relation to immobilization efficiency, Zymomonas mobilis showed high adhesion and colonization in support, indicated by cell growth increased from 107 to 10(9) CFU mL(-1) during fermentation time.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)