Behavioral and neurochemical effects on rat offspring after prenatal exposure to ethanol.

The work studied behavioral and neurochemical alterations in 21-day-old pups, from both sexes (26 g on average) born from female Wistar rats administered daily with ethanol (0.5 or 4.0 g/kg, p.o.), for 30 days before mating, and throughout their gestational period. Ethanol administration continued from delivery up to weaning. The open field, elevated plus maze and forced swimming tests were used to evaluate effects of ethanol on locomotion, anxiety and depression, respectively. Binding assays were used to identify dopaminergic (D1- and D2-like) and muscarinic (M1 plus M2) receptors. Results of the plus maze test indicated significant and dose-dependent increases in the number of entrances in the open arms and in the time of permanence in the open arms, in the prenatally ethanol-exposed offspring, as compared to controls, indicating an anxiolytic effect. In the open field test, this group presented decreases in spontaneous locomotor activity as well as in the occurrences of rearing and grooming. Offspring also showed dose-dependent increases in their immobility time in the forced swimming test, characterizing despair behavior. Decreases in the hippocampal (D2: 32%; D1: 25%) and striatal (D2: 30%; D1: 52%) dopaminergic binding were detected in ethanol-exposed offspring. On the other hand, significant increases were observed in muscarinic binding in the hippocampus (40%) as well as in the striatum (42%). This study shows evidence that in utero ethanol exposure produces a long-lasting effect on development and pharmacological characteristics of brain systems that may have important implications in behavioral and neurochemical responsiveness occurring in adulthood.

Determination of 21-hydroxydeflazacort in human plasma by high-performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry. Application to bioequivalence study.

A liquid chromatographic atmospheric pressure chemical ionization tandem mass spectrometric method is described for the determination of 21-hydroxydeflazacort in human plasma using dexamethasone 21-acetate as an internal standard. The procedure requires a single diethyl ether extraction. After evaporation of the solvent under a nitrogen flow, the analytes are reconstituted in the mobile phase, chromatographed on a C18 reversed-phase column and analyzed by mass spectrometry via a heated nebulizer interface where they are detected by multiple reaction monitoring. The method has a chromatographic run time of less than 5 min and a linear calibration curve with a range of 1-400 ng ml(-1) (r>0.999). The between-run precision, based on the relative standard deviation for replicate quality controls, was < or =5.5% (10 ng ml(-1)), 1.0% (50 ng ml(-1)) and 2.7% (200 ng ml(-1)). The between-run accuracy was +/-7.1, 3.8 and 4.8% for the above concentrations, respectively. This method was employed in a bioequivalence study of two DFZ tablet formulations (Denacen from Marjan Industria e Comercio, Brazil, as a test formulation, and Calcort from Merrell Lepetit, Brazil, as a reference formulation) in 24 healthy volunteers of both sexes who received a single 30 mg dose of each formulation. The study was conducted using an open, randomized, two-period crossover design with a 7-day washout interval. The 90% confidence interval (CI) of the individual geometric mean ratio for Denacen/Calcort was 89.8-109.5% for area under the curve AUC(0-24 h) and 80.7-98.5% for Cmax. Since both the 90% CI for AUC(0-24 h) and Cmax were included in the 80-125% interval proposed by the US Food and Drug Administration, Denacen was considered bioequivalent to Calcort according to both the rate and extent of absorption.

Pharmacokinetics of deflazacort in renal transplanted and hemodialyzed children.

Deflazacort (DFZ) pharmacokinetics was evaluated in fifteen pediatric patients on chronic hemodialysis or after renal transplantation, and in three normal children. After overnight fasting, oral DFZ 0.26+/-0.01 mg/kg (mean +/- SEM) was given. Serial blood samples were collected for 360 min and analyzed by HPLC for 21-hydroxy-DFZ (21-HO-DFZ). Serum concentration profiles and pharmacokinetic parameters were similar in patients on hemodialysis, renal transplant recipients and normal children. Elimination half-life was longer in the 9 cyclosporine-treated subjects (108.0+/-13.6 min) than in the other nine (71.2+/-8.3 min; p <0.02). Our finding suggests that, from a pharmacokinetic point of view, DFZ dose adjustment for renal function is not necessary in children with chronic renal failure or after renal transplantation.

Binding of the anti-inflammatory steroid deflazacort to glucocorticoid receptors in brain and peripheral tissues. In vivo and in vitro studies.

Deflazacort (DFC) is a heterocyclic glucocorticoid with anti-inflammatory activity but with decreased side effects. In this study, we have evaluated the capacity of DFC and other glucocorticoids to reach the central nervous system (CNS) in vivo by measuring changes of [3H]dexamethasone (DEX) binding to glucocorticoid receptors (GR) in vitro. GR occupation was effected by DEX in the cerebral cortex, hippocampus, pituitary, liver and thymus, with DFC showing a similar profile except for the cerebral cortex. In contrast, corticosterone weakly occupied GR in the thymus, pituitary and hippocampus and methyl-prednisolone was active only in peripheral tissues. Furthermore, IC50 for DEX in vitro amounted to 15-17 nM in the hippocampus and liver, whereas IC50 for the active metabolite 21-deacetyl-DFC (21-OH-DFC) was 4 times higher. 21-OH-DFC bound to type II and was absent from type I GR. When tested in equipotent doses based on IC50 analysis, DFC and DEX similarly induced in vivo ornithine decarboxylase activity in hippocampus and liver, although body weight loss after chronic treatment was significantly less for DFC. The results show that DFC distributes on the CNS similarly to DEX, induces ornithine decarboxylase activity but presents less intensive catabolic effects, making it suitable for use as an anti-inflammatory steroid during chronic therapeutic regimes.