Determination of quorum-sensing signal substances in water and solid phases of activated sludge systems using liquid chromatography–mass spectrometry

The detection of acyl homoserine lactones（AHLs） in activated sludge is essential for clarifying their function in wastewater treatment processes. An LC–MS/MS method was developed for the detection of AHLs in both the aqueous and solid phases of activated sludge. In addition, the effects of proteases and extracellular polymeric substances（EPS） on the detection of AHLs were evaluated by adding protease inhibitors and extracting EPS,respectively. Recoveries of each AHL were improved by adding 50 μL of protease inhibitor,and recoveries were also improved from 0 to 56.9% to 24.2%–105.8% by EPS extraction.Applying the developed method to determine the type and concentration of AHLs showed that C4-HSL, C6-HSL, C8-HSL and 3-oxo-C8-HSL were widely detected in a suspended activated sludge system. The dominant AHL was C8-HSL, with a highest concentration of304.3 ng/L. C4-HSL was mainly distributed in the aqueous phase, whereas C6-HSL, C8-HSL and 3-oxo-C8-HSL were preferentially distributed in the sludge phase.

The early signal substances induced by heat stress in brains of mice

To study the effects of early signal substances induced by heat stress in brains of Kunming mice,six-month-old mice(n 572)were pretreated with heat stress and subsequent ischemia/reperfusion by clipping of their bilateral cervical common arteries for 7 min.According to different treatments,animals were randomly divided into four groups:(1)normal control group;(2)heat stress pre-treatment followed by ischemia and reperfusion group(HS/IR);(3)ischemia and reperfusion group(IR);(4)heat stress group(HS).Animals in the later three groups were subdivided into 3 subgroups(1 day,4 days,14 days),respectively.The changes in the expression of cAMP res-ponse element binding protein(CREB)and calcitonin gene-related peptide(CGRP)were detected by immuno-histochemistry and computer image analysis methods.The results showed that compared with the normal group,the expressions of CREB in the hippocampal CA1 region increased significantly in the HS,HS/IR and IR groups(P,0.05).Compared to the normal group,heat stress could result in CGRP excretion and redistribution in the cerebrum,with the highest level in the 4 d HS/IR group.Following heat stress,CGRP immunoreactivity was observed in varicose fibers and neuronal perikarya within the CA1 region.The results indicate that heat stress can induce CREB expression,which in turn stimulates CGRP secretion.