Nicotine-Induced Effects on Nicotinic Acetylcholine Receptors (nAChRs), Ca2+ and Brain-Derived Neurotrophic Factor (BDNF) in STC-1 Cells

In addition to the T2R bitter taste receptors, neuronal nicotinic acetylcholine receptors (nAChRs) have recently been shown to be involved in the bitter taste transduction of nicotine, acetylcholine and ethanol. However, at present it is not clear if nAChRs are expressed in enteroendocrine cells other than beta cells of the pancreas and enterochromaffin cells, and if they play a role in the synthesis and release of neurohumoral peptides. Accordingly, we investigated the expression and functional role of nAChRs in enteroendocrine STC-1 cells. Our studies using RT-PCR, qRT-PCR, immunohistochemical and Western blotting techniques demonstrate that STC-1 cells express several α and β nAChR subunits. Exposing STC-1 cells to nicotine acutely (24h) or chronically (4 days) induced a differential increase in the expression of nAChR subunit mRNA and protein in a dose- and time-dependent fashion. Mecamylamine, a non-selective antagonist of nAChRs, inhibited the nicotineinduced increase in mRNA expression of nAChRs. Exposing STC-1 cells to nicotine increased intracellular Ca2+ in a dose-dependent manner that was inhibited in the presence of mecamylamine or dihydro-β-erythroidine, a α4β2 nAChR antagonist. Brain-derived neurotrophic factor (BDNF) mRNA and protein were detected in STC-1 cells using RT-PCR, specific BDNF antibody, and enzyme-linked immunosorbent assay. Acute nicotine exposure (30 min) decreased the cellular content of BDNF in STC-1 cells. The nicotine-induced decrease in BDNF was inhibited in the presence of mecamylamine. We also detected α3 and β4 mRNA in intestinal mucosal cells and α3 protein expression in intestinal enteroendocrine cells. We conclude that STC-1 cells and intestinal enteroendocrine cells express nAChRs. In STC-1 cells nAChR expression is modulated by exposure to nicotine in a doseand time-dependent manner. Nicotine interacts with nAChRs and inhibits BDNF expression in STC-1 cells

Toxicopathological changes on Wistar rat after multiple exposures to acetamiprid

Abstract Aim: To see the toxicopathological changes after multiple exposure to acetamiprid (ACP) and also to obtain more information regarding the manner in which ACP acts at cellular level. Materials and Methods: A subacute toxicity study of ACP was undertaken in 72 female Wistar rats in four groups (18 each). Three different concentrations of ACP (25, 100 and 200 mg/kg of body weight) were administered orally to rats. Untreated rats served as control. Different plasma enzyme and analytes were measured. Gross and histopathological observations were noted in this experiment. Result: There was a significant increase in the plasma enzymes tested in this experiment. There was a significant decrease in plasma glucose, cholesterol and low-density lipid. Necrotic and degenerative changes were observed in vital organs. Conclusion: It is observed that ACP has the toxic potential (on liver, kidney, heart, ovary and brain) at sub lethal doses

Cyclic-AMP regulates postnatal development of neural and behavioral responses to NaCl in rats

During postnatal development rats demonstrate an age-dependent increase in NaCl chorda tympani (CT) responses and the number of functional apical amiloride-sensitive epithelial Na+channels (ENaCs) in salt sensing fungiform (FF) taste receptor cells (TRCs). Currently, the intracellular signals that regulate the postnatal development of salt taste have not been identified. We investigated the effect of cAMP, a downstream signal for arginine vasopressin (AVP) action, on the postnatal development of NaCl responses in 19–23 day old rats. ENaC-dependent NaCl CT responses were monitored after lingual application of 8-chlorophenylthio-cAMP (8-CPT-cAMP) under open-circuit conditions and under ±60 mV lingual voltage clamp. Behavioral responses were tested using 2 bottle/24h NaCl preference tests. The effect of [deamino-Cys1, D-Arg8]-vasopressin (dDAVP, a specific V2R agonist) was investigated on ENaC subunit trafficking in rat FF TRCs and on cAMP generation in cultured adult human FF taste cells (HBO cells). Our results show that in 19–23 day old rats, the ENaC-dependent maximum NaCl CT response was a saturating sigmoidal function of 8-CPT-cAMP concentration. 8-CPT-cAMP increased the voltage-sensitivity of the NaCl CT response and the apical Na+ response conductance. Intravenous injections of dDAVP increased ENaC expression and γ-ENaC trafficking from cytosolic compartment to the apical compartment in rat FF TRCs. In HBO cells dDAVP increased intracellular cAMP and cAMP increased trafficking of γ- and δ-ENaC from cytosolic compartment to the apical compartment 10 min post-cAMP treatment. Control 19–23 day old rats were indifferent to NaCl, but showed clear preference for appetitive NaCl concentrations after 8-CPT-cAMP treatment. Relative to adult rats, 14 day old rats demonstrated significantly less V2R antibody binding in circumvallate TRCs. We conclude that an age-dependent increase in V2R expression produces an AVP-induced incremental increase in cAMP that modulates the postnatal increase in TRC ENaC and the neural and behavioral responses to NaCl

Release of GLP-1 and PYY in response to the activation of G protein-coupled bile acid receptor TGR5 is mediated by Epac/PLC-ε pathway and modulated by endogenous H2S

Activation of plasma membrane TGR5 receptors in enteroendocrine cells by bile acids is known to regulate gastrointestinal secretion and motility and glucose homeostasis. The endocrine functions of the gut are modulated by microenvironment of the distal gut predominantly by sulfur-containing bacteria of the microbiota that produce H2S. However, the mechanisms involved in the release of peptide hormones, GLP-1 and PYY in response to TGR5 activation by bile acids and the effect of H2S on bile acid-induced release of GLP-1 and PYY are unclear. In the present study, we have identified the signaling pathways activated by the bile acid receptor TGR5 to mediate GLP-1 and PYY release and the mechanism of inhibition of their release by H2S in enteroendocrine cells. The TGR5 ligand oleanolic acid (OA) stimulated Gs and cAMP formation, and caused GLP-1 and PYY release. OA-induced cAMP formation and peptide release were blocked by TGR5 siRNA. OA also caused an increase in PI hydrolysis and intracellular Ca2+. Increase in PI hydrolysis was abolished in cells transfected with PLC-ε siRNA. 8-pCPT-2’-O-Me-cAMP, a selective activator of Epac, stimulated PI hydrolysis, and GLP-1 and PYY release. L-Cysteine, which activates endogenous H2S producing enzymes cystathionine--lyase and cystathionine--synthase, and NaHS and GYY4137, which generate H2S, inhibited PI hydrolysis and GLP-1 and PYY release in response to OA or 8-pCPT-2’-O-Me-cAMP. Propargylglycine, an inhibitor of CSE, reversed the effect of L-cysteine on PI hydrolysis and GLP-1 and PYY release. We conclude: i) activation of Gs-coupled TGR5 receptors causes stimulation of PI hydrolysis, and release of GLP-1 and PYY via a PKA-independent, cAMP-dependent mechanism involving Epac/PLC-/Ca2+ pathway, and ii) H2S has potent inhibitory effects on GLP-1 and PYY release in response to TGR5 activation, and the mechanism involves inhibition of PLC-/Ca2+ pathway

The Simulations Chain of the MURAVES Experiment

The MUon RAdiography of VESuvius (MURAVES) project is aimed at studying the summital cone of Mt. Vesuvius, an active and hazardous volcano near Naples, Italy. A detailed Monte Carlo simulation framework is necessary in order to investigate the effects of the experimental constraints and to perform comparisons with the actual observations. Our Monte Carlo setup combines a variety of Monte Carlo programs that address different aspects of cosmic muon simulation, from muon generation in the Earth’s upper atmosphere to the response of the detector, including the interactions with the material of the volcano. We will elaborate on the rationale for our technical choices, including the trade-off between speed and accuracy, and on the lessons learned, which are of general interest for similar use cases in muon radiograph

Effect of an Adjacent Flat Plate on a Highly-Heated Rectangular Supersonic Jet

Solid surfaces located in the vicinity of a supersonic jet may affect its flow dynamics and greatly change its aeroacoustic characteristics. Large-eddy simulations (LES) are performed to investigate the effects of a plate on a highly-heated rectangular supersonic jet. The rectangular nozzle has an aspect ratio of 2.0 and is operated at overexpanded conditions with a nozzle pressure ratio of 3.0 and a nozzle temperature ratio of 7.0. Four cases with a plate-to-nozzle distance ranging from 0 to 3 times of the jet equivalent nozzle diameter are investigated. The large-scale implicit LES computations are performed by a well-validated in-house finite-volume based CFD code, which uses an artificial dissipation mechanism to represent the effects of small-scale turbulence structures and to damp the numerical oscillations near shocks. The temperature-dependent thermal properties of air in the highly-heated jets are considered by the chemical equilibrium assumption. Numerical results show that among the four jets, the case with the plate directly attached at the nozzle lip shows significant different flow and acoustic fields from the others. It exhibits a longer jet potential core length but without forming a series of well-structured shock diamonds. The other cases show similar shock/expansion wave structures as observed in the free jet but their jet plumes bend towards the plate. This bending leads to one jet to scrub over the plate in the downstream. The scrubbing effect, together with the unaffected shock-shear layer interactions and high plate pressure loading, leads to a stronger acoustic power in the near acoustic fields for this jet as compared to the others. The spectrum analysis in the nozzle upstream direction shows that the plate removes or mitigates the screech tone observed in the free jet and slightly amplifies the acoustic amplitudes in the low-frequency range

Complete genome sequence of Mycobacterium phlei type strain RIVM601174

Item does not contain fulltextMycobacterium phlei is a rapidly growing nontuberculous Mycobacterium species that is typically nonpathogenic, with few reported cases of human disease. Here we report the whole genome sequence of M. phlei type strain RIVM601174.1 juni 201

The MURAVES Experiment: A Study of the Vesuvius Great Cone with Muon Radiography

The MURAVES experiment aims at the muographic imaging of the internal structure of the summit of Mt. Vesuvius, exploiting muons produced by cosmic rays. Though presently quiescent, the volcano carries a dramatic hazard in its highly populated surroundings. The challenging measurement of the rock density distribution in its summit by muography, in conjunction with data from other geophysical techniques, can help the modeling of possible eruptive dynamics. The MURAVES apparatus consists of an array of three independent and identical muon trackers, with a total sensitive area of 3 square meters. In each tracker, a sequence of 4 XY tracking planes made of plastic scintillators is complemented by a 60 cm thick lead wall inserted between the two downstream planes to improve rejection of background from low-energy muons. The apparatus is currently acquiring data. Preliminary results from the analysis of the first data sample are presented

CD200 Receptor Controls Sex-Specific TLR7 Responses to Viral Infection

Immunological checkpoints, such as the inhibitory CD200 receptor (CD200R), play a dual role in balancing the immune system during microbial infection. On the one hand these inhibitory signals prevent excessive immune mediated pathology but on the other hand they may impair clearance of the pathogen. We studied the influence of the inhibitory CD200-CD200R axis on clearance and pathology in two different virus infection models. We find that lack of CD200R signaling strongly enhances type I interferon (IFN) production and viral clearance and improves the outcome of mouse hepatitis corona virus (MHV) infection, particularly in female mice. MHV clearance is known to be dependent on Toll like receptor 7 (TLR7)-mediated type I IFN production and sex differences in TLR7 responses previously have been reported for humans. We therefore hypothesize that CD200R ligation suppresses TLR7 responses and that release of this inhibition enlarges sex differences in TLR7 signaling. This hypothesis is supported by our findings that in vivo administration of synthetic TLR7 ligand leads to enhanced type I IFN production, particularly in female Cd200−/− mice and that CD200R ligation inhibits TLR7 signaling in vitro. In influenza A virus infection we show that viral clearance is determined by sex but not by CD200R signaling. However, absence of CD200R in influenza A virus infection results in enhanced lung neutrophil influx and pathology in females. Thus, CD200-CD200R and sex are host factors that together determine the outcome of viral infection. Our data predict a sex bias in both beneficial and pathological immune responses to virus infection upon therapeutic targeting of CD200-CD200R