In recent years, a new class of designer drugs has appeared on the drugs of abuse market in many countries, namely, the so-called beta-keto (bk) designer drugs such as mephedrone (bk-4-methylmethamphetamine), butylone (bk-MBDB), and methylone (bk-MDMA). The aim of the present study was to identify the metabolites of mephedrone in rat and human urine using GC-MS techniques and to include mephedrone, butylone, and methylone within the authors' systematic toxicological analysis (STA) procedure. Six phase I metabolites of mephedrone were detected in rat urine and seven in human urine suggesting the following metabolic steps: N-demethylation to the primary amine, reduction of the keto moiety to the respective alcohol, and oxidation of the tolyl moiety to the corresponding alcohols and carboxylic acid. The STA procedure allowed the detection of mephedrone, butylone, methylone, and their metabolites in urine of rats treated with doses corresponding to those reported for abuse of amphetamines. Besides macro-based data evaluation, an automated evaluation using the automated mass spectral deconvolution and identification system was performed. Mephedrone and butylone could be detected also in human urine samples submitted for drug testing. Assuming similar kinetics in humans, the described STA procedure should be suitable for proof of an intake of the bk-designer drugs in human urine.

Deletions a ecting the interval between the RB1 gene and marker D13S25 at band 13q14 are the most frequent genetic abnormalities of B-cell chronic lymphocytic leukemia (B-CLL) and indicate the presence of a novel tumor suppressor gene in this region. In the current study, a high resolution physical map of fragments spanning one megabasepair (Mb) of genomic DNA at the critical 13q14 segment was constructed. To de®ne the minimal region of loss within the RB1 and D13S25 interval, we screened 322 B-CLLs for deletions at either of the two loci. Thirty mantle cell lymphomas (MCLs) were included in the analysis because we observed a 13q14 deletion pattern similar to B-CLL in this disease. The incidence of 13q14 deletions was 51% in B-CLL and 70% in MCL, respectively. No frequent loss of the BRCA2 gene at band 13q12 was found. Detailed deletion mapping at band 13q14 with probes from the RB1 ± D13S25 interval lead to the identi®cation of a critical deletion region 400 kb in size. Within this region two segments were most frequently a ected, one at D13S272 120 kb in size and another 240 kb distal of D13S272 80 kb in size. From these two segments expressed sequences were identi®ed as candidates for the putative 13q14 tumor suppressor gene involved in the pathogenesis of B-CLL and MCL.Keywords: deletion mapping; FISH; exon ampli®cation; DIRVISH The molecular pathogenesis of B-CLL, the most frequent leukemia in Western countries, is not well understood (Rozman and Montserrat, 1995). Although a number of recurrent chromosome aberrations have been identi®ed, no gene to date has been found to be consistently involved (Juliusson et al., 1990;DoÈ hner et al., 1997a).The most common structural chromosome aberrations in B-CLL involve band 13q14 (Juliusson et al., 1990), to which the RB1 tumor suppressor gene has been localized (Friend et al., 1986). Monoallelic RB1 deletion was frequently observed in B-CLL but disruption of both RB1 alleles occurred rarely (Liu et al., 1992(Liu et al., , 1993Stilgenbauer et al., 1993). Based on the observation that the distal marker D13S25 was deleted more frequently than RB1, a novel B-CLL tumor suppressor gene located in vicinity to this marker was postulated (Brown et al., 1993;Liu et al., 1993;Chapman et al., 1994).Subsequent studies aimed at the localization of this gene have been thus far inconclusive. Devilder et al.(1995) localized a minimal deletion segment distal of D13S25. However, comparing deletions of RB1 and D13S25 lead us to the hypothesis that the critical segment is localized proximal of D13S25 (Stilgenbauer et al., 1995). This was supported by Liu et al. (1995) who found loss of D13S319 located less than 680 kb proximal of D13S25 more frequently deleted than other 13q14 markers. Bullrich et al. (1996) recently assembled a YAC contig to order new markers in the region and identi®ed a minimal deletion between D13S25 and the marker 206XF12 located less than 550 kb proximal of D13S25. However, in this study deletions did not appear to cluster in a single critical region and the poten...

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