Viability and cell cycle analysis of equine fibroblasts cultured in vitro

This experiment aimed to study equine fibroblasts in culture analyzing and the cell cycle and viability of cells pre- and post-freezing. Skin fragments were obtained from 6 horses and cultured in DMEM high glucose + 10% FCS in 5% CO(2) until the beginning of confluence. Two passages were performed before freezing. Cells subjected to serum starvation (0.5% FCS) were analyzed for viability and cell cycle at 24, 48, 72, 96, 120, 144 and 168 h of culture. For the confluent groups, cells were analyzed at the moment they achieved confluence. Cellular viability was assisted with Hoescht 33342 and propidium iodide. The analysis of apoptosis/necrosis and cell cycle was performed using a flow cytometer (FACS Calibur BD(A (R))) after staining the cells with annexin V and propidium iodide. Both optical microscopy and flow cytometry confirmed that cellular viability was similar for serum starvation and confluent groups (average 84%). Similarly, both methods were efficient to synchronize the cell cycle before freezing. However, after thawing, serum starvation, for more than 24 h, was superior to culture for synchronizing cells in G0/G1 (69% x 90%). The results of this experiment indicate that equine fibroblasts can be efficiently cultured after thawing.

The Effects of N Helix Deletion and Mutant F29W on the Ca2+ Binding and Functional Properties of Chicken Skeletal Muscle Troponin C

To assess the structural and functional significance of the N helix (residues 3-13) of avian recombinant troponin C (rTnC), we have constructed NHdel, in which residues 1-11 have been deleted, both in rTnC and in the spectral probe mutant F29W (Pearlstone, J. R., Borgford, T., Chandra, M., Oikawa, K., Kay, C. M., Herzberg, O., Moult, J., Herklotz, A., Reinach, F. C., and Smillie, L.B. (1992) Biochemistry 31, 6545-6553). Comparison of the far- and near-UV CD spectra (±Ca2+) of F29W and F29W/ NHdel and titration of the Ca2+-induced ellipticity and fluorescence changes indicates that the deletion has little effect on the global fold of the molecule but reduces the Ca2+ affinity of the N domain, but not the C domain, by 1.6-1.8-fold. Comparisons of the mutants NHdel, F29W, and F29W/NHdel with rTnC have been made using several functional assays. In reconstituted troponin-tropomyosin actomyosin subfragment 1 and myofibrillar ATPase systems, both F29W and NHdel have significantly reduced Ca2+-activated enzymic activities. These effects are cumulative in the double mutant F29W/ NHdel. On the other hand, maximal isometric tension development in Ca2+-activated reconstituted skinned fibers is not affected with F29W and NHdel, although the Ca2+ sensitivity of NHdel in this system is markedly reduced. We conclude that both mutations, NHdel and F29W, are functionally deleterious, possibly affecting interactions of the N domain with troponin I and/or T.

COX24 codes for a mitochondrial protein required for processing of the COX1 transcript

In most strains of Saccharomyces cerevisiae the mitochondrial gene COX1, for subunit 1 of cytochrome oxidase, contains multiple exons and introns. Processing of COX1 primary transcript requires accessory proteins factors, some of which are encoded by nuclear genes and others by reading frames residing in some of the introns of the COX1 and COB genes. Here we show that the low molecular weight protein product of open reading frame YLR204W, for which we propose the name COX24, is also involved in processing of COX1 RNA intermediates. The growth defect of cox24 mutants is partially rescued in strains harboring mitochondrial DNA lacking introns. Northern blot analyses of mitochondrial transcripts indicate cox24 null mutants to be blocked in processing of introns aI2 and aI3. The dependence of intron aI3 excision on Cox24p is also supported by the growth properties of the cox24 mutant harboring mitochondrial DNA with different intron compositions. The intermediate phenotype of the cox24 mutant in the background of intronless mitochondrial DNA, however, suggests that in addition to its role in splicing of the COX1 pre-mRNA, Cox24p still has another function. Based on the analysis of a cox14-cox24 double mutant, we propose that the other function of Cox24p is related to translation of the COX1 mRNA. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

Sensory profile and acceptability of traditional and double-distilled cachaca aged in oak casks

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

The contribution of genes required for anaerobic respiration to the virulence of Salmonella enterica serovar Gallinarum for chickens

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

Electrophoretic and chromatographic profile for S-like hemoglobin

Hemoglobin variants originate mainly by simple amino acid substitutions, the result of nucleotide sequence changes. Recently, the number of known abnormal hemoglobins has increased due to improvement in analysis methodologies; however, many laboratories are not prepared to correctly identify mutants. Hb S is a very well-characterized hemoglobin variant that varies in prevalence in different regions of Brazil. However, there is a type of Hb that presents electrophoretic migration in alkaline pH similar to Hb S, named S-like Hb, which can be incorrectly diagnosed; therefore, its frequency is underestimated. We obtained reference ranges for Hb S by HPLC, and we examined the electrophoretic and chromatographic profiles of S-like Hb. Hb Hasharon, Hb D-Los Angeles, Hb Korle-Bu, Hb Lepore, Hb D-Iran, Hb G-like, Hb Queens, Hb Montgomery, and Hb Q-India were found. Cases of association between two beta chain mutants were also found. Electrophoresis in alkaline and acid pH was utilized to initially screen these Hb variants, and globin chain electrophoresis at both high and low pH was performed to identify the globin chain mutant. Chromatographic analysis permitted the identification of the hemoglobin variant and also facilitated the quantification of these variants. Therefore, an association of classical laboratory diagnostic methodologies is fundamental for the correct identification of suspect Hb variants. The S and S-like hemoglobin profiles determined in this study will help in the diagnosis of these variants in health care services.

Environmental evaluation of cogeneration scenarios in sugar industry

The development of new techniques that allow the analysis and optimization of energy systems bearing in mind environmental issues is indispensable in a world with finite natural resources and growing demand of energy. Among the energy systems that deserve special attention, cogeneration in the sugar industry must be pointed out, because it uses efficiently a common fuel for generation of useful heat and power. Within this frame, thermoeconomical optimization - 2nd Law of Thermodynamics analysis by exergy function and economic evaluation of the thermal system - gradually is taking importance as a powerful tool to assist to the decision making process. Also, the explicit consideration of environmental issues offers a better way to explore trade-offs between different aspects to support the decisions that must be made. In this work it is used the technique of Life Cycle Analysis (LCA) which allows to consider environmental matters as an integral part of the problem, in opposite to most of the environmental approaches that only reduce residuals generation , without taking into account impacts associated to other related processes. On the other hand, the consideration of environmental issues in optimization of energy systems is a novel and promissory contribution in the state of the art of energy optimization and LCA. The system under study is a sugar plant of Tucumán (Argentina) given the particular importance that this industry had inside the regional economy of the Argentinean Northwest. Although cogeneration comes being used a while ago in sugar industry, being the main objective the generation of heat and as secondary objective the electric power generation and mechanic power to cover several needs of working machineries, to the date it is no available a versatile tool that allows to analyze economical feasible alternatives bearing in mind environmental issues. At sugar plants, steam is generated in boilers using as fuel bagasse - cellulosic fiber waste obtained crushing the sugar cane- and it is used to give useful heat and shaft work to the plant, but it can also be used to generate electricity with export opportunities to the electrical network. The great number of process alternatives outlines a serious decision making problem in order to take advantage of the resources. Although the problem turns out to be a mixed non-linear problem (MINLP), the main contribution of this work is the development of a hybrid strategy to evaluate cogeneration alternatives that combines optimization approaches with environmental indicators. This powerful tool for its versatility and robustness to analyze cogeneration systems, will be of great help in the decision making process, because of their easy implementation to analyze the kind of problems presented in the sugar industry.

Efeito da bidestilação na qualidade sensorial da cachaça

Pós-graduação em Alimentos e Nutrição - FCFAR

Estudo de viabilidade de implantação, operação e monitoramento de aterros sanitários: uma abordagem econômica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Osteotomia com brocas de alta e baixa rotação, com e sem refrigeração líquida: estudo histopatológico duplo cego em suínos

Pós-graduação em Odontologia - FOA

Papel funcional de microRNAs na arquitetura vegetativa e radicular de plantas

Pós-graduação em Ciências Biológicas (Genética) - IBB

Subsídios para avaliação do ciclo de vida de modo simplificada da produção agrícola de milho por meio de um estudo de caso

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise da viabilidade e ciclo celular de fibroblastos equinos cultivados in vitro: comparação pré e pós-descongelação

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aplicabilidade da metodologia ecodesign à produção de calçados femininos

Pós-graduação em Design - FAAC

Genistein at Maximal Physiologic Serum Levels Induces G0/G1 Arrest in MCF-7 and HB4a Cells, But Not Apoptosis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)