[Trypanosoma cruzi: obtaining extracellular amastigotes and studying their development under different culture conditions]. / Trypanosoma cruzi: obtención de amastigotes extracelulares y estudio de su crecimiento en diferentes condiciones de cultivo.

This work describes a protocol to obtain pure populations of extracellular amastigotes of Trypanosoma cruzi. The amastigote stage was obtained by means of temperature changes and human plasma added to the culture medium. Epimastigotes (clon BraC15C2) were first grown in F69 medium at 27 degrees C during 96 h and then at 36.5 degrees C. After three subcultures of 96 h each at the latter temperature a subsequent incubation in the presence of 5% human plasma, was needed to obtain a population of amastigotes that could be maintained indefinitely in the F69 or F29 media. This amastigote population was similar morphologically to that obtained through other methods. The kinetic of growth depended on the culture medium used (F29 or Brain-Heart Infusion, BHI). When culture was incubated at 27 degrees C in both media, the pre-exponential and logarithmic phases of growth were observed at 72-96 h and 24-48 h respectively. The change in stage from amastigote to epimastigote dependent whether amastigote were subcultured or not. The growth of amastigotes in BHI medium at 36.5 degrees C did not occurred. The growth of amastigotes was similar to those observed at 27 degrees C when F29 medium was used although the transformation to epimastigotes did not take place at this temperature. A population over 99% of amastigotes were maintained at 36.5 degrees C indefinitely by means of subcultures in F29 medium.

Effect of arsenic (V) on the antioxidant defense system: in vitro oxidation of rat plasma lipoprotein.

Adult male rats were treated orally with sodium arsenate (10 mg As/kd/day) for 2 days, and in increase in hepatic glutathione level was seen. Ascorbic acid content increased in both liver and plasma of intoxicated animals. Hepatic activities of superoxide dismutase and glutathione peroxidase did not change with the treatment and there was no increase in the level of lipid peroxidation measured as thiobarbituric acid-reacting substances (TBARS). Arsenic decreased the plasma level of uric acid and increased the plasma triglycerides content without modifying vitamin E levels. Both total lipoproteins and very low density lipoprotein plus low density lipoprotein (VLDL + LDL) fractions demonstrated greater propensity for in vitro oxidation than the corresponding untreated rats. The last finding might be a useful parameter for determining the degree of oxidative stress in the initial steps of intoxication with arsenic.

The pharmacokinetics of ceftazidime in lactating and non-lactating cows.

The pharmacokinetics of ceftazidime (CAZ) were studied in lactating (LTG) and non-lactating (NLTG) cows. Two groups (LTG and NLTG) of 5 healthy dairy cows were given ceftazidime (10 mg/ kg body weight) intravenously (i.v.) and intramuscularly (i.m.). Serum and milk (LTG) and serum samples (NLTG) were collected over a 24-h period post-administration. CAZ concentrations in serum and milk were determined by high-performance liquid chromatography, and an interactive and weighted-non-linear least-squares regression analysis was used to perform the pharmacokinetic analysis. The pharmacokinetic profiles in LTG and NLTG cows which had received CAZ i.v. fitted a three-compartment model and a two-compartment model, respectively. The CAZ concentration-time curves in serum and the area under the curve were greater and more sustained (p < 0.05) in the LTG cows by both routes, while the serum clearance (Cls = 72.5 +/- 18.1 ml/h per kg) was lower (p < 0.05) than that in the NLTG cows (Cls = 185.9 +/- 44.2 ml/h per kg). CAZ given i.v. exhibited a relatively long half-life of elimination (t1/2 beta (LTG) = 1.1 +/- 0.2 h; t1/2 beta (NLTG) = 1.4 +/- 0.3 h). Compared with other cephalosporins, CAZ had good penetration into the mammary gland (47.7 +/- 38.2% for CAZ i.v.; 51.1 +/- 39.0% for CAZ i.m.). Finally, the bioavailability of CAZ (F(LTG) = 98.9 +/- 36.8%; F(NLTG) = 77.1 +/- 25.3%) was suitable for its used by the i.m. route in lactating and non-lactating cows.