24 resultados para Free Culture

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


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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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The synthesis of polygalacturonases (PG) is known to be influenced by Aspergillus growth conditions, namely, environmental factors and pectin content in the cultivation medium containing a mixed carbon source. Optimal conditions were attained at a temperature of 30 A degrees C and an initial pH of 4.5. PG activity (3.29 and 2.48 U/mL) was determined after a two-day culture of Aspergillus sp. HC1 and Aspergillus sp. CC1, respectively, in a basic medium containing 2% citrus pectin as the sole carbon source. The addition of glucose (2% w/v) to the basic medium led to a 2-fold increase in PG production. However, enzyme synthesis was repressed when a higher concentration of glucose was used in the medium containing the mixed carbon source. Spores from the two fungi were immobilized in a 3% Ca-alginate system and the mechanical strength of the gel beads allowed the use of this process system 6-fold longer (288 h) than the free culture. In the Aspergillus sp. CC1 immobilized system, PG production increased nearly 10-fold in the medium with 2% glucose added (5.95 U/mL) in comparison to the medium without sugar (0.55 U/mL). The results demonstrate that a different response in activity was produced by free and entrapped spore systems. PG production remained approximately constant throughout the six 48 h cycles in the medium containing citrus pectin (2% w/v) as the sole carbon source.

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Objective: The present study evaluated the cytotoxic effects of hard setting applied on the odontoblastlike cells MDPC-23. Study design: Eighty round-shaped samples were prepared with the following experimental materials: calcium hydroxide, Vitrebond, RelyX Luting, and RelyX Unicem. The samples were placed in serum-free culture medium and incubated for 24 hours or 7 days at 37°C with 5% CO 2 and 95% air. The odontoblast cells were plated in the wells and incubated for 72 hours. After this period, the complete culture medium was replaced by the extracts obtained from every sample, and the methyltetrazolium assay was carried out to evaluate the cell metabolism. Results: For the 24-hour period, the experimental materials calcium hydroxide, Vitrebond, RelyX Luting, and RelyX Unicem decreased the cell metabolic activity by 91.52%, 81.14%, 78.17%, and 2.64%, respectively. For the 7-day period, calcium hydroxide, Vitrebond, RelyX Luting, and RelyX Unicem decreased the metabolic activity of the MDPC-23 cells by 91.13%, 87.27%, 79.04%, and 10.51%, respectively. Conclusion: RelyX Unicem presented the lowest cytopathic effects to the cultured odontoblast cell line. © 2007 Mosby, Inc. All rights reserved.

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The major challenge for a greater dissemination of in vitro produced (IVP) bovine embryos is to improve embryonic survival after cryopreservation. The involvement of embryonic lipids on this issue is well documented. However, it has been recognized that not only the amount of lipids that affects embryo cryotolerance, but the embryo survival capacity after cryopreservation is a rather multifactorial event. In this review, some strategies to improve embryonic lipid composition and postcryopreservation survival by modifying the embryos themselves to make them more cryopreservable are overviewed. The use of semi-defined and defined serum-free culture media, the addition of some chemicals in the culture media to modify embryo lipid composition, and the modulation of embryo cell membrane fluidity by cholesterol or unsaturated fatty acids added to the culture media and oocyte/embryo donor nutritional management with a diet enriched in polyunsaturated fatty acids, were described as alternatives for the improvement of IVP embryo survival after cryopreservation.

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This study evaluated the effects of LLLT on the expressionof inflammatory cytokines related to the development of oralmucositis by gingival fibroblasts. Primary gingival fibroblastswere seeded on 24-well plates (105cells/well) for 24 h. Freshserum-free culture medium (DMEM) was then added, andcells were placed in contact with LPS (Escherichia coli,1 lgmL1), followed by LLLT irradiation (LaserTABLE—InGaAsP diode prototype—780 nm, 25 mW) delivering 0,0.5, 1.5 or 3 J cm². Cells without contact with LPS werealso irradiated with the same energy densities. Gene expres-sion of TNF- a, IL-1b, IL-6 and IL-8 was evaluated by Real-Time PCR, and protein synthesis of these cytokines wasdetermined by enzyme-linked immunosorbent (ELISA) assay.Data were statistically analyzed by the Kruskal– Wallis test,complemented by the Mann–Whitney test (P < 0.05). LPStreatment increased the gene expression and protein synthesisof TNF-a, IL-6 and IL-8, while the expression of IL-1b wasnot affected. For LPS-treated groups, LLLT promoted signif-icant decreases in the expression of TNF-a, IL-6, and IL-8 at1.5 J cm2and 3 J cm2. These results demonstrate thatLLLT promoted a beneficial biomodulatory effect on theexpression of inflammatory cytokines related to oral mucosi-tis by human gingival fibroblasts.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Pós-graduação em Microbiologia Agropecuária - FCAV

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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

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The mostly binucleate trophoblast giant cells (TGC) found in bovine placentomes, in addition to synthesizing and releasing hormones play an important role in fetal development and maternal adaptation to pregnancy. Placentomes from early gestation were collected, and for isolation of mature TGC, three cellular disaggregation methods, mechanical (MECH), enzymatic by trypsin (TRYP) or collagenase (COLL) were compared to each other. Further on, the cell survival in culture medium (DMEM) supplemented with either 10% fetal calf serum (FCS) or 10% serum replacement (SR) on culture plates free of any substrate was evaluated over a period of 90 days by trypan blue exclusion. The cells were further characterized by HOECHST 33342 nuclear staining, and immunocytochemical staining with monoclonal antibodies against vimentim and cytokeratin. A mean total rate of TGC survival of 82.56% was recorded. Statistical analysis showed significantly higher survival rates after enzymatic disaggregation with COLL (86.23%) than following MECH (80.38%) or TRYP (80.91%) treatment. Supplementation of DMEM with FCS resulted in significantly higher cellular survival rates (87.13%) when compared to the addition of SR (77.73%). Analysis of the influence of both, disaggregation method and medium supplementation on TGC survival revealed statistically significant differences between the following groups: MECH-SR (71.09%) was significantly lower than all other groups; TRYP-SR (78.03%) was significantly different from all other groups; TRYP-FCS (83.43%) and COLL-SR (84.08%) were significantly lower than MECH-FCS (89.98%) which together with COLL-FCS (88.25%) showed the highest cellular survival rate. In summary, our results show that TGC isolated from early gestation placentomes may be viable for more than 90 days of culture. However, whether these TGC produce placental lactogen throughout this period has yet to be determined. (c) 2006 Elsevier B.V. All rights reserved.

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The production of extracellular cellulase-free xylanase from Trichoderma inhamatum was evaluated in liquid Vogel medium with different carbon sources as natural substrates and agricultural or agro-industrial wastes. Optimal production of 244.02 U/mL was obtained with xylan as carbon source, pH 6.0 at 25 degrees C, 120 rpm, and 60-h time culture. Optimal conditions for enzyme activity were 50 degrees C and pH 5.5. Thermal stability of T. inhamatum xylanolytic complex expressed as T(1/2) was 2.2 h at 40 degrees C and 2 min at 50 degrees C. The pH stability was high from 4.0 to 11.0.These results indicate possible employment of such enzymatic complex in some industrial processes which require activity in acid pH, wide-ranging pH stability, and cellulase activity absence.

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The objective of the study was to evaluate the effect of different concentrations of nitrogen and phosphorus on the growth of the free-floating aquatic macrophytes Eichhornia crassipes, Pistia stratiotes, and Salvinia molesta. The plants were cultured in 2,000-l outdoor concrete tanks. Triplicate tanks, with a continuous flow of effluent from culture ponds containing Nile tilapia, were used for each plant type (n = 3). The plant material was collected monthly from 0.25 m(2) floating quadrats, at the two ends of the tanks (higher nutrient concentrations near the inflow and lower nutrient concentrations near the outflow). In low nutrient concentrations, the maximum relative growth rates (RGRs) for E. crassipes (0.016/day) and P. stratiotes (0.016/day) were significantly lower (P <= 0.05) than for S. molesta (0.029/day). There were no significant differences between the RGRs of S. molesta in the different nutrient concentrations. Eichhornia crassipes and P. stratiotes had their growth limited by nitrogen and phosphorus concentrations. The increase in plant density during the experiment probably also affected the growth of these species. In this context, E. crassipes and P. stratiotes can cause problems in nutrient-rich waterbodies, but under these experimental conditions their growth was limited by nitrogen and/or phosphorus concentrations. The growth of S. molesta was not influenced by the different nutrient concentrations.