Ethylglucuronide determination in urine and hair from alcohol withdrawal patients.

Two methods for the determination of ethylglucuronide (EtG) in urine and in hair have been developed by liquid chromatography- tandem mass spectrometry. These two methods were fully validated, including linearity (0.25-100 microg/mL in urine; 0.05-5 ng/mg in hair; r(2) > 0.99, n = 5), limits of detection (0.1 microg/mL in urine, 0.025 ng/mg in hair) and quantitation (lowest level of the calibration curve), extraction efficiency (> 55%), within-day and between-day imprecision and bias (CV and mean relative error < 15%), matrix effect, and relative ion intensity. These methods have been applied to 541 urine samples and 17 hair specimens collected from 156 alcohol withdrawal patients. The determination of ethanol versus EtG in urine was compared, and also the convenience of EtG determination in hair. EtG in urine and in hair proved to be a powerful tool for monitoring abstinence in these patients.

A novel indazolo-triazolo-benzotriazepine exerts anti-inflammatory effects by inhibition of cyclooxygenase-2 activity and nitric oxide synthase-2 expression.

We have studied the anti-inflammatory activity of (6-(p-bromophenyl)amino-7-(p-chlorophenyl)indazolo[2',3':1,5]-1,2,4-triazolo[4,3-a]-1,3,5-benzotriazepine (ITB), prepared by solid-phase synthesis. This novel compound reduced the production of nitrite and PGE(2) in RAW 264.7 macrophages stimulated with lipopolysaccharide in a concentration-dependent manner. The first effect was dependent on inhibition of nitric oxide synthase-2 (NOS-2) protein expression although ITB did not modify nuclear factor-kappa B (NF-kappa B)-DNA binding. In addition, this compound inhibited cyclooxygenase-2 (COX-2) activity, which was also observed in aspirin-treated human monocytes. The anti-inflammatory effects of ITB were demonstrated in the carrageenan-induced mouse paw oedema, where this compound inhibited swelling and PGE(2) levels in inflamed paws but not in stomachs, in contrast to the dual COX-1/COX-2 inhibitor indomethacin. Inhibition of COX-2 activity and NOS-2 protein expression was confirmed in vivo using the zymosan-injected mouse air pouch model. These results suggest that synthesis of fused indazolo bis(guanidines) is a useful approach in the search for new anti-inflammatory agents.