Impaired ethanol metabolism can lead to various alcohol-related health problems. Key enzymes in ethanol metabolism are alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH); however, neuroendocrine pathways that regulate the activities of these enzymes are largely unexplored. Here we identified a neuroendocrine system involving Corazonin (Crz) neuropeptide and its receptor (CrzR) as important physiological regulators of ethanol metabolism in Drosophila. Crz-cell deficient (Crz-CD) flies displayed significantly delayed recovery from ethanol-induced sedation that we refer to as hangover-like phenotype. Newly generated mutant lacking Crz Receptor (CrzR01) and CrzR-knockdown flies showed even more severe hangover-like phenotype, which is causally associated with fast accumulation of acetaldehyde in the CrzR01 mutant following ethanol exposure. Higher levels of acetaldehyde are likely due to 30% reduced ALDH activity in the mutants. Moreover, increased ADH activity was found in the CrzR01 mutant, but not in the Crz-CD flies. Quantitative RT-PCR revealed transcriptional upregulation of Adh gene in the CrzR01. Transgenic inhibition of cyclic AMP-dependent protein kinase (PKA) also results in significantly increased ADH activity and Adh mRNA levels, indicating PKA-dependent transcriptional regulation of Adh by CrzR. Furthermore, inhibition of PKA or cAMP response element binding protein (CREB) in CrzR cells leads to comparable hangover-like phenotype to the CrzR01 mutant. These findings suggest that CrzR-associated signaling pathway is critical for ethanol detoxification via Crz-dependent regulation of ALDH activity and Crz-independent transcriptional regulation of ADH. Our study provides new insights into the neuroendocrine-associated ethanol-related behavior and metabolism.

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We investigated the use of amniocentesis performed at eight to 14 weeks' gestation as a possible alternative to chorionic villus sampling. Patients, methods, and results Samples of amniotic fluid were taken from 40 gestation, and the mean time to the cells being harvested was 12 6 days. In contrast only 17 (68%) of the 25 samples taken at eight to 11 weeks yielded a result. One sample taken at 13 weeks' gestation yielded a female karyotype, whereas the fetal parts revealed a male karyotype; the sample was subsequently identified as maternal urine. The mean volume of amniotic fluid obtained was 13 9 ml (range 1-40 ml). CommentAll 15 samples taken at 12-14 weeks' gestation yielded a result. The mean time to cells being harvested in this group (12-6 days) compared favourably with the current mean of 11 days for the samples obtained routinely at [16][17][18][19] weeks that are processed by our laboratory. Culture of all the 5 ml aliquots obtained at 12-14 weeks was successful. Thus a 10 ml sample would provide two cultures, which are necessary for the interpretation of equivocal results and in case of microbial infection.In one case, a urine sample was obtained at 13 weeks' gestation from an obese patient in whom imaging was poor. In a clinical environment sampling would not have been attempted, and this patient would have been recalled later.Our results show that amniocentesis from as early as 12 weeks' gestation can provide sufficient material for cytogenetic diagnosis and could be offered as an alternative to current methods of prenatal diagnosis. Furthermore, the procedure could be carried out by doctors already familiar with the technique, using existing resources. Patients must, however, be advised that the risks of this procedure are unknown. Preliminary reports from the United States suggest that early amniocentesis is safer than chorionic villus sampling.24 Further evaluation, preferably by means of a randomised trial, is urgently needed. We are continuing our investigation of amniocentesis before 12 weeks with the aim of bringing the procedure forward into the first trimester of pregnancy.We acknowledge contributions to the study from Mr N Fisk, Mr P Reginald, Mr M Michel, and Mrs R Rebello.

Summary Trophoblast was obtained by ordinary suction curettage and by transcervical aspiration with a medicut cannula from women having a therapeutic abortion in the first trimester of pregnancy. The decidual tissue which is invariably attached to early placental villi was separated and pure cultures obtained from the trophoblast layers and from the mesenchymal core of placental villi. Cytotrophoblast had a very limited life span in tissue culture, whereas mesenchymal cells grew rapidly and could be used for antenatal diagnosis.

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