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Construção e caracterização cinética e fisiológica de um sistema células Sf9/ baculovírus recombinante para a produção de Canacistatina
(Universidade Federal de São Carlos, 2007-03-14) Arantes, Mabel Karina; Suazo, Cláudio Alberto Torres; https://lattes.cnpq.br/9591447226240450; https://lattes.cnpq.br/4495308167335518
The development of the biotechnology has permitted to achieve, among other biotechnological products, many recombinant proteins which are largely used as vaccines, hormones, enzymes and other ways of human therapeutic activity. This is possible because the multidisciplinary: through the technology of the recombinant DNA the region that encode gene of interest is cloned in a system of adequate expression for super-expression of the correspondent protein and, through the technology of bioprocess is possible to increase the obtaining scale of the recombinant product assuring its reproducibility, integrity and functionality. Involving these two areas of knowledge, this works intends to express an heterolog protein, the Canacistatina, deriving from sugar cane, through an eucariotic expression system Sf9/baculovirus. This implies in the construction of a recombining baculovirus, containing the cDNA which codifies to the Canacistatina, and also the characterization of the cultivation of Sf9 cells, and of the process of infection by baculovirus, in kinetics and physiologic therms. In order to obtain these purposes it was done experiments of cultivation of Sf9 cells in Schott bottles of 100 mL, with work volume of 20 mL, in 27° C. In these experiments it was analyzed the optimum conditions of velocity agitation, inoculum s density, culture environment, and also the role of substrates. In relation to these parameters, it was observed that a better cell growth can be conduced in a 100 rpm of agitation, in an inoculum s density of 1x106 cell/ml, using a combination of 1:1 between the mediums SF900 II and TNM-FH (Medium 50%), without the necessity of addition of Bovine Fetal Sorum. This last aspect represents a great advantage in relation to the bioprocesses, in a way that it uses Sf9 cells already known. It was construed the recombining baculovírus containing the Canacistatina s gene through the modified genome of the baculovirus Autographa californica Multiple Nuclear Polihedrosis Virus and the infection experiments of Sf9 cells was made under three MOI values (1, 5 e 10 pfu/cell) in the two culture mediums. The betters results indicates to infection of Sf9 cells in 50% Medium using MOI= 5 pfu/cell, conditions that the Canacistatina s productivity reached 28 µg/106cel.
Detecção do limiar de lactato salivar e da variabilidade da freqüência cardíaca em exercício resistido: comparação entre métodos
(Universidade Federal de São Carlos, 2007-03-14) Oliveira, João Carlos de; Perez, Sergio Eduardo de Andrade; https://lattes.cnpq.br/9408953831211309; https://lattes.cnpq.br/7208666542619125
Important researches had shown the benefits of resistance training programs for rehabilitation as well as prophylaxis of physical incapacity. However few studies had investigated the responses of the intermediate metabolism during the execution of resistance exercises if carried through steady state or during incremental tests. The purpose of this study was to investigate the viability of the use of non-invasive methods as trustworthy indicators in the analysis of the aerobic-anaerobic transition and to compare the physiological responses between male and female, submitted to discontinuous incremental test of resistance exercise for lower limb. Eighteen subjects volunteered to participate and provided a written consent (9 males, 24 ± 3 years, 176 ± 5 cm, 72 ± 10 kg and 9 females, 23 ± 2 years, 165 ± 6 cm, 59 ± 10 Kg) in two sessions, distinct days, of counterweighted isoinertial Leg Press 45º machine (LP), being the 1st session performed to identify the 1 repetition maximum load (1RM) and the 2nd to accomplish the discontinuous incremental test of the type step in LP (DIT). The loading gradual scheme was determined in relation to the percentage of 1RM (%1RM). Subjects were required to perform 20 ± 2 repetitions with a 2 minutes interval between each set when loads were changed and collection of blood and salivary samples were collected. The blood lactate concentration ([La-]) had been used as gold standard for the respective comparisons between the methods. As non-invasive methods the salivary lactate concentrations ([SL]) and index SD1 of the heart rate variability (HRV) were used, gotten by Poincaré Plotting (PP). The thresholds identified by [La-] and [SL] (LT and SLT, respectively) had been determined by Mader and Heck Method (1986). The threshold identified by index SD1 (SD1T) was determined by Tullpo et al. (1998) criterion. The criterion level for statistical significance was set at an alpha level of p< 0,05. Statistical differences had not been found in the comparisons between genders in each step of the DIT for the three variables studied. The thresholds when expressed in kg revealed significant differences between genders but not intra genders. When expressed in %1RM the thresholds had been placed in the average enter 29-40% and had not evidenced significant differences between the thresholds in the conditions intra genders and between genders. Correlations of 0.89 and 0.78 had been evidenced between the LT and SLT and SD1T, respectively. The variability of both non-invasive variables was shown inside of the limits of agreement evidenced by the analyses of Bland-Altman. Thus it is possible to conclude that as much the SL as index SD1 of HRV responses of similar form to the blood lactate. This allows the detention of the thresholds of lactate salivary and heart rate variability, independently of the gender. In this way, as much the SL as index SD1 of the HRV presents great potential as possible non invasive markers of effort intensity, that they can be used in the lapsing and the control of the load of work in the resistance exercises. Moreover, the differences in the metabolic responses between the genders, evidenced in the intensities of effort above of the threshold, must be considered in the lapsing of the resistance training.