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.

No abstract

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.

No abstract

Early amniocentesis between 11 and 14 weeks' gestation was offered to 110 women at risk of a chromosomally abnormal fetus due to maternal age. Four were found to be unsuitable for the procedure, and 106 early amniocenteses were performed. In 102 cases, clear amniotic fluid was obtained with a single tap. There were two dry taps and two bloodstained taps; sampling was repeated in three of these cases before 15 weeks. In the fourth case, placental biopsy was performed at 16 weeks. Thus, we were able to obtain a satisfactory sample in all but three cases (2.8 per cent). Karyotyping of cells harvested from the early amniotic fluid samples was successful in all the 105 cases. Cell culture from the initial samples revealed a normal karyotype in 99 cases, two balanced translocations, two tetraploid karyotypes, and two cases of pseudomosaicism. Of the 105 pregnancies successfully sampled, there have been two losses to date (1.8 per cent). Two further patients presented with premature rupture of membranes, both pregnancies having successful outcomes. Sixty-two babies have delivered to date, with four congenital anomalies. There were no respiratory problems. Twenty-nine pregnancies are continuing without known complications, and details are not yet available on the remaining 12. The results indicate that early amniocentesis may replace the traditional test at 15-17 weeks.

One hundred and fourteen samples of amniotic fluid taken before 15 weeks of gestation were cultured for cytogenetic studies. The results of culturing these early amniotic fluid (EAF) samples were compared with the results of culturing 114 standard amniotic fluid (SAF) samples taken after 15 weeks of gestation matched for maternal age and received in the laboratory within the same week. Cell culture was successful in all 114 of the EAF samples and in 111 SAF samples. There was no significant difference in the days to harvesting and days to reporting in the two groups. Three samples of SAF failed to grow and two EAF samples produced tetraploid karyotypes, so that in these five cases amniocentesis had to be repeated. These problems were attributed to toxicity of a fungicide used in the culture medium. Pseudo-mosaicism was noted in two EAF samples and one SAF sample; and maternal cell contamination was noted in one EAF and one SAF sample. Thus, culturing and karyotyping cells harvested from EAF and SAF are similar, indicating that EAF samples from 12-14-week pregnancies could be used for prenatal diagnosis.

As the frequency and intensity of extreme events such as droughts, heatwaves and floods have increased over recent decades, more extreme biological responses are being reported, and there is widespread interest in attributing such responses to anthropogenic climate change. However, the formal detection and attribution of biological responses to climate change is associated with many challenges. We illustrate these challenges with data from the Elbe River floodplain, Germany. Using community turnover and stability indices, we show that responses in plant, carabid and mollusc communities are detectable following extreme events. Community composition and species dominance changed following the extreme flood and summer heatwave of 2002/2003 (all taxa); the 2006 flood and heatwave (molluscs); and after the recurring floods and heatwave of 2010 and the 2013 flood (plants). Nevertheless, our ability to attribute these responses to anthropogenic climate change is limited by high natural variability in climate and biological data; lack of long-term data and replication, and the effects of multiple events. Without better understanding of the mechanisms behind change and the interactions, feedbacks and potentially lagged responses, multiple-driver attribution is unlikely. We discuss whether formal detection and/or attribution is necessary and suggest ways in which understanding of biological responses to extreme events could progress. Extreme climatological events are important drivers associated with ongoing anthropogenic climate change 1,2. As mean climate conditions change, the frequency and intensity of extreme events such as droughts, heatwaves and floods are also projected to increase 3. Extreme events can result in changes to the distribution of populations of individual species or community-level responses such as changes to species richness, composition and/or dominance e.g. 4-6. These changes may be long lasting or irreversible if competitive interactions are altered, especially when species become (locally) extinct, or with recurring extreme events e.g. 7-10. Extreme biological responses to individual extreme weather events are already being observed in many ecosystems around the world 9,11-17 , and interest in attributing such responses to anthropogenic climate change is increasing 1. However, it has been questioned whether it is possible (or indeed necessary) to formally detect and attribute biological responses to anthropogenic climate change (henceforth "climate change") as is done in the climate system 18, 19. The IPCC defines "detection" as a demonstration that the likelihood of occurrence of an observed change is significantly different from that due to natural internal variability, without attempting to explain the causes of the observed change 20,21. In contrast, "attribution" attempts to identify the most likely causes for the detected change with some defined level of confidence. Attribution requires that the detected change is