Simultaneous detection of autophagy and apoptosis is important in drug discovery and signaling studies. Here we report, a real-time reporter cell line for the simultaneous detection of apoptosis and autophagy at single-cell level employing stable integration of two fluorescent protein reporters of apoptosis and autophagy. Cells stably expressing EGFP-LC3 fusion was developed initially as a marker for autophagy and subsequently stably expressed with inter-mitochondrial membrane protein SMAC with RFP fusion to detect mitochondrial permeabilization event of apoptosis. The cell lines faithfully reported the LC3 punctae formation and release of intermembrane proteins in response to diverse apoptotic and autophagic stimuli. Risk assessment for mixtures of chemicals requires to investigate the magnitude of their potential adverse effects on living organisms. This is usually done by assessing how experimental toxicological mixture data depart from the model of Loewe additivity.  https://www.selleckchem.com/products/GDC-0980-RG7422.html  Several recent scientific studies propose to perform this task using an ad hoc method known as model deviation ratio (MDR) method. Moreover, the first official European regulatory document for the study of combined exposures explicitly recommends the use of the MDR method (EFSA Scientific Committee et al. Guidance on harmonised methodologies for human health, animal health and ecological risk assessment of combined exposure to multiple chemicals. EFSA Journal, 2019). We show here that the MDR method is not rooted in statistical principles and can lead to erroneous claims. We show however that the distribution of the MDR can be evaluated by simulations and show how this allows us to devise and carry out a bona fide statistical test. The proposed method accounts for uncertainty in the estimation of ED/EC50 and does not require a minimum sample size. The computer code developped in this study is made available as an R package called MDR. The possibility that exposure to inhalation anaesthetics inhibits neurogenesis and results in memory deficits has attracted considerable interest over the past decade. This study was designed to investigate the mechanism of the sevoflurane exposure-induced decline in hippocampal neurogenesis. Young mice were anaesthetized with a gaseous mixture of 3.0% sevoflurane/60% oxygen 2 h daily for three consecutive days. Sodium butyrate (NaB) administration began 2 h prior to anaesthesia and continued daily until the end of behavioural tests. The Morris water maze (MWM) test was used to determine spatial learning and memory performance. We assessed the effect of repeated sevoflurane exposure on histone acetylation and the expression of brain-derived neurotropic factor (BDNF) and its receptor, tropomyosin-related kinase receptor B (TrkB), in the hippocampus by Western blot (WB). To detect neurogenesis, we first counted the number of neural stem cells (NSCs); we then assessed their proliferation level by immunohistochemistry and estimated the number of new-born cells by immunofluorescence. We found that sevoflurane induced learning and memory deficits in young mice 4 weeks after sevoflurane exposure and that NaB injection restored histone acetylation and improved the performance of the mice in the MWM. NaB also increased the number and proliferation of NSCs and neonatal cells, which were inhibited by sevoflurane. Concomitantly, BDNF and TrkB expression, which was decreased by sevoflurane, was also restored by NaB. Our study showed that sevoflurane affects long-term neurocognitive function and neurogenesis in young mice. Normalization of histone acetylation may alleviate the neurodevelopmental side effects of this anaesthetic. Tuning of the cardiovascular response is crucial to maintain performance during high-intensity exercise. It is well known that the nucleus of the solitary tract (NTS) in the brainstem medulla plays a central role in cardiovascular regulation; however, where and how upper brain regions form circuits with NTS and coordinately control cardiovascular responses during high-intensity exercise remain unclear. Here focusing on the amygdala and claustrum, we investigated part of the mechanism for regulation of the cardiovascular system during exercise. In rats, c-Fos immunostaining was used to examine whether the amygdala and claustrum were activated during treadmill exercise. Further, we examined arterial pressure responses to electrical and chemical stimulation of the claustrum region exhibiting exercise intensity-dependent activation. We also confirmed the anatomical connections between the amygdala, claustrum, and NTS by retrograde tracer injections. Finally, we performed simultaneous electrical stimulation of the claustrum and amygdala to examine their functional connectivity. c-Fos expression was observed in the amygdala and the posterior part of the claustrum (pCL), but not in the anterior part, in an exercise intensity-dependent manner. pCL stimulation induced a depressor response. Using a retrograde tracer, we confirmed direct projections from the amygdala to the pCL and NTS. Simultaneous stimulation of the central nucleus of the amygdala and pCL showed a greater pressor response compared with the stimulation of the amygdala alone. These results suggest the amygdala and pCL are involved in different phases of exercise. More speculatively, these areas might coordinately tune cardiovascular responses that help maintain performance during high-intensity exercise. The mesopontine tegmental anesthesia area (MPTA) is a small brainstem nucleus that, when exposed to minute quantities of GABAA receptor agonists, induces a state of general anesthesia. In addition to immobility and analgesia this state is accompanied by widespread suppression of neural activity in the cerebral cortex and high delta-band power in the electroencephalogram. Collectively, MPTA neurons are known to project to a variety of forebrain targets which are known to relay to the cortex in a highly distributed manner. Here we ask whether ascending projections of individual MPTA neurons collateralize to several of these cortical relay nuclei, or access only one. Using rats, contrasting retrograde tracers were microinjected pairwise on one side into three ascending relays the basal forebrain, the zona incerta-lateral hypothalamus and the intralaminar thalamic nuclear group. In addition, in separate animals, each target was microinjected bilaterally. MPTA neurons were then identified as being single-or double-labeled, indicating projection to one target nucleus or collateralization to both.