Ruling out pyridine dinucleotides as true TRPM2 channel activators reveals novel direct agonist ADP-ribose-2'-phosphate

Transient receptor potential melastatin 2 (TRPM2), a Ca(2+)-permeable cation channel implicated in postischemic neuronal cell death, leukocyte activation, and insulin secretion, is activated by intracellular ADP ribose (ADPR). In addition, the pyridine dinucleotides nicotinamide-adenine-dinucleotide (NAD), nicotinic acid-adenine-dinucleotide (NAAD), and NAAD-2'-phosphate (NAADP) have been shown to activate TRPM2, or to enhance its activation by ADPR, when dialyzed into cells. The precise subset of nucleotides that act directly on the TRPM2 protein, however, is unknown. Here, we use a heterologously expressed, affinity-purified-specific ADPR hydrolase to purify commercial preparations of pyridine dinucleotides from substantial contaminations by ADPR or ADPR-2'-phosphate (ADPRP). Direct application of purified NAD, NAAD, or NAADP to the cytosolic face of TRPM2 channels in inside-out patches demonstrated that none of them stimulates gating, or affects channel activation by ADPR, indicating that none of these dinucleotides directly binds to TRPM2. Instead, our experiments identify for the first time ADPRP as a true direct TRPM2 agonist of potential biological interest

Novel ionization reagent for the measurement of gas-phase ammonia and amines using a stand-alone atmospheric pressure gas chromatography (APGC) source.

RationaleContaminants present in ambient air or in sampling lines can interfere with the target analysis through overlapping peaks or causing a high background. This study presents a positive outcome from the unexpected presence of N-methyl-2-pyrrolidone, released from a PALL HEPA filter, in the analysis of atmospherically relevant gas-phase amines using chemical ionization mass spectrometry.MethodsGas-phase measurements were performed using a triple quadrupole mass spectrometer equipped with a modified atmospheric pressure gas chromatography (APGC) source which allows sampling of the headspace above pure amine standards. Gas-phase N-methyl-2-pyrrolidone (NMP) emitted from a PALL HEPA filter located in the inlet stream served as the ionizing agent.ResultsThis study demonstrates that some alkylamines efficiently form a [NMP + amine+H]+ cluster with NMP upon chemical ionization at atmospheric pressure. The extent of cluster formation depends largely on the proton affinity of the amine compared with that of NMP. Aromatic amines (aniline, pyridine) and diamines (putrescine) were shown not to form cluster ions with NMP.ConclusionsThe use of NMP as an ionizing agent with stand-alone APGC provided high sensitivity for ammonia and the smaller amines. The main advantages, in addition to sensitivity, are direct sampling into the APGC source and avoiding uptake on sampling lines which can be a significant problem with ammonia and amines

Effect of spray cooling on heat transfer in a two-phase helium flow

International audienceIn this paper, we describe an experimental study of the phenomenon of spray cooling in the case of liquid helium, either normal or superfluid, and its relationship to the heat transfer between an atomized diphasic flow contained in a long pipe, and the pipe walls. This situation is discussed in the context of the cooling of the superconducting magnets of the Large Hadron Collider (LHC). Experiments were conducted in a test loop reproducing the LHC cooling system, in which the vapor velocity and temperature could be varied in a large range. Shear induced atomization results in the generation of a droplet mist which was characterized by optical means. The thin liquid film deposited on the walls by the mist was measured using interdigitated capacitors. The cooling power of the mist was measured using thermal probes, and correlated to the local mist density. Analysis of the results shows that superfluidity has only a limited influence on both the film thickness and the mist cooling power. Using a simple model, we show that the phenomenon of spray cooling accounts for the measured non linearity of the global heat transfer. Finally, we discuss the relevance of our results for cooling the final focus magnets (inner triplets) in an upgraded version of the LHC