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Protein and trypsin activity of each fraction (2 mL) were established according to Warburg and Christian [18] and Freitas-Júnior et al. [2], respectively.
Levels of mutational load (ML) were established based on the frequency with which a particular level of ML was observed in microdissected targets histologically classified as intestinal metaplasia.
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On the contrary, when decreasing the current from I = 620 mA, the second harmonic ML is established first, but limited in the left by an abrupt jump to the fundamental branch at I = 425 mA, indicating that the state with a pair of equal-spaced pulses propagating in the cavity is no longer a solution for this ML system at such low current.
The NJ tree was obtained using the routine in MEGA 4 [ 21] MP analyses were performed using PAUP* beta 10 version [ 22] and ML reconstructions were established using PhyML 2.4.4 [ 23].
NOZ cells in log-phase growth were resuspended in serum-free culture medium (at a density of 1 × 10 cells in 0.2 mL), and then tumor xenografts were established by subcutaneous inoculation of these NOZ cells into the right flank of nude mice.
The bacterial stock cultures were established in 200 ml Erlenmeyer flasks under constant aeration in the dark at room temperature (15°C) and fresh nutrient broth was added once a week during 2 weeks cultivation.
Cultures were established in 3 ml of growth medium and cells were allowed to attach.
A most remarkable observation was made in that the favourable prognostic effect of the female gender was restricted to patients aged less than 60 years (ms 13.3 mo vs 10.1 mo; 2ys 26% vs 5%), whereas for older women no advantages over men's results were established (ms 9.3 ml vs 9.1 mo; 2ys 8% vs 7%).
Chemostats with individually calibrated pH and temperature probes were established in 500-mL fermenter vessels (Sixfors; Infors AG, Bottmingen, Switzerland) containing 300-mL cultures.
Stable clones were selected with use of puromycin (1 µg/ml); and after they were established, they were screened for expression levels of Chk2 protein relative to control.
Thus, the following 4 categories were established: negative (<22 pg/mL), low (22 99 pg/mL), medium (100 999 pg/mL) and high (≥1000 pg/mL).
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