DMPP reduced nitrification,but not annual N2O emissions from mineral fertilizer applied to oilseed rape on a sandy loam soil

Direct field emissions of nitrous oxide (N₂O) may determine whether biodiesel from oilseed rape (Brassica napus L.) fulfills the EU requirement of at least 50% reduction of greenhouse gas emissions as compared to fossil diesel. However, only few studies have documented fertilizer N emission factors (EF) and mitigation options for N₂O emissions from oilseed rape cropping systems. We conducted a field experiment with three N levels (0, 171, and 217 kg/ha), where the N fertilizer was applied as ammonium sulfate nitrate with or without the nitrification inhibitor 3,4‐dimethylpyrazole phosphate (DMPP). N₂O fluxes were measured using static chambers technique and soil samples were analyzed for water and mineral N content during a monitoring period of 368 days. The DMPP treatments showed a significantly increased level of ammonium () for up to 18 weeks after spring fertilization as compared to the treatments without DMPP. However, this difference did not result in a corresponding decrease in soil content, and no differences in cumulative N₂O emissions were found between any fertilized treatments with or without DMPP (mean, 1.26 kg N₂O‐N ha^?1 year^?1). More field experiments are needed to clarify whether DMPP‐coated mineral fertilizers could mitigate N₂O emissions under different weather conditions, for example, under conditions where fertilization events concurred with rainfall events increasing water‐filled pore space to the assumed 60% threshold for denitrification. Emission factors for mineral N fertilizer were 0.28%–0.36% with a mean of 0.32% across the fertilized treatments. These data concur with recent European studies suggesting that the EF for mineral N fertilizers in oilseed rape cropping systems may typically be lower than the default IPCC value of 1%. Further studies are needed to consolidate an EF for oilseed rape under temperate conditions, which will be determining for the sustainability of Northern European oilseed rape cultivation for biodiesel.     

Phosphorylation of tyrosine hydroxylase in the superior cervical ganglion

We studied the phosphorylation of tyrosine hydroxylase in the superior cervical ganglion of the rat. Ganglia were preincubated with [³²P]P_i and were then incubated in non-radioactive medium containing a variety of agents that are known to activate tyrosine hydroxylase in this tissue. Tyrosine hydroxylase was isolated from homogenates of the ganglia by immunoprecipitation followed by polyacrylamide gel electrophoresis. ³²P-labelled tyrosine hydroxylase was visualized by radioautography, and the incorporation of ³²P into the enzyme was quantitated by densitometry of the autoradiograms. Veratridine produced a concentration-dependent increase in the incorporation of ³²P into tyrosine hydroxylase, with 50 μM veratridine producing a 5-fold increase in ³²P incorporation. The nicotinic agonist, dimethylphenylpiperazinium (100 μM), caused a 7-fold increase in the phosphorylation of tyrosine hydroxylase. The effect of dimethylphenylpiperazinium was maximal within 1 min and decreased upon continued exposure of the ganglia to this agent. The actions of dimethylphenylpiperazinium and of veratridine were dependent on extracellular Ca²⁺. Muscarine, 8-Br-cAMP, forskolin, vasoactive intestinal peptide, isoproterenol, deoxycholate and phospholipase C also stimulated the incorporation of ³²P into tyrosine hydroxylase. These data support the hypothesis that phosphorylation plays a role in activation of tyrosine hydroxylase produced by all of these agents.     

Dimethyl phenyl piperazine iodide (DMPP) induces glioma regression by inhibiting angiogenesis

1,1-Dimethyl-4-phenyl piperazine iodide (DMPP) is a synthetic nicotinic acetylcholine receptor (nAChR) agonist that could reduce airway inflammation. In this study, we demonstrated that DMPP could dramatically inhibit glioma size maintained on the chick embryonic chorioallantoic membrane (CAM). We first performed MTT and BrdU incorporation experiments on U87 glioma cells in vitro to understand the mechanism involved. We established that DMPP did not significantly affect U87 cell proliferation and survival. We speculated that DMPP directly caused the tumor to regress by affecting the vasculature in and around the implanted tumor on our chick CAM model. Hence, we conducted detailed analysis of DMPP's inhibitory effects on angiogenesis. Three vasculogenesis and angiogenesis in vivo models were used in the study which included (1) early chick blood islands formation, (2) chick yolk-sac membrane (YSW) and (3) CAM models. The results revealed that DMPP directly suppressed all developmental stages involved in vasculogenesis and angiogenesis – possibly by acting through Ang-1 and HIF-2α signaling. In sum, our results show that DMPP could induce glioma regression grown on CAM by inhibiting vasculogenesis and angiogenesis.     

Differential Effect of Nicotinic Agonists on the [3H]Norepinephrine Release from Rat Hippocampal Slices

The aim of this study was to investigate the mechanisms involved in the effect of nicotinic agonists on the [³H]norepinephrine ([³H]NE) release from rat hippocampal slices. The stimulatory effect of nicotine, cytisine, epibatidine and anatoxin-A was completely blocked by the nicotinic antagonist mecamylamine (10 M). In contrast, the effect of dimethylphenylpiperazinium (DMPP) was only partially inhibited by mecamylamine but was completely blocked by the NE uptake inhibitor desipramine (DMI, 10 M). Finally, the effect of lobeline was not affected by mecamylamine and was only partially blocked by DMI. Our data indicate that the majority of nicotinic agonists increase the release of [³H]NE exclusively via stimulation of nicotinic acetylcholine receptors (nAChRs). DMPP, in addition to the stimulation of nAChRs, also evokes a carrier-mediated release. Lobeline has no stimulatory effect on nAChRs, induces a carrier-mediated release and has a further action of unidentified mechanism. Our results suggest that special caution is required for the interpretation of data, when DMPP or lobeline are used as nicotinic agonists.     

突触前α7烟碱受体对海马神经元兴奋性突触传递的调控

采用盲法膜片钳技术观察突触前烟碱受体(nicotinic acetylcholinel receptors,nAChRs)对海马脑片CAl区锥体神经元兴奋性突触传递的调控作用。结果显示,nAChRs激动剂碘化二甲基苯基哌嗪(dimethylphenyl—piperazinium iodide,DMPP)不能在CAl区锥体神经元上诱发出烟碱电流。DMPP对CAl区锥体神经元自发兴奋性突触后电流(spontaneous excitatory postsynaptic current,sEPSC)具有明显的增频和增幅作用,并呈现明显的浓度依赖关系。DMPP对微小兴奋性突触后电流(miniature excitatory postsynaptic current,mEPSC)具有增频作用,但不具有增幅作用。上述DMPP增强突触传递的作用不能被nAChRs拮抗剂美加明、六烃季铵和双氢-β-刺桐丁所阻断,但可被α-银环蛇毒素阻断。上述结果提示,海马脑片CAl区锥体神经元兴奋性突触前nAChRs含有对α-银环蛇毒素敏感的胡亚单位,其激活可增强海马CAl区锥体神经元突触前递质谷氨酸的释放,从而对兴奋性突触传递发挥调控作用。     

Ammonium Nutrition as a Strategy for Cadmium Mobilisation in the Rhizosphere of Sunflower

Ammonium nutrition of higher plants results in rhizosphere acidification due to proton excretion by root cells. The acidification induced by ammonium-fed plants can be exploited to promote a localised metal mobilisation in neutral to alkaline polluted soils and therefore to improve phytoextraction. The effects of ammonium uptake by sunflower (Helianthus annuus L.) plants on the external medium pH, aerial and root growth and tolerance to soluble Cd were studied in hydroponic culture. The ammonium-fed sunflowers induced a strong acidification of the solution and, compared to the nitrate-fed sunflowers, a small modification in mineral nutrition and a different Cd partitioning between root and shoot. Moreover, ammonium nutrition was found to induce a great mobilisation of a sparingly soluble form of cadmium (CdCO₃). A pot experiment studied the ability of different ammonium-based fertilisers (ammonium sulphate, ammonium thiosulphate, urea) to modify bulk and rhizo-soil pH, compared to the effect of calcium nitrate and to the unfertilised soil. Furthermore, in order to promote the persistence of ammonium in soil, a combined treatment of ammonium sulphate and DMPP, a nitrification inhibitor, was tested. Soil pH was strongly modified by chemical and biological processes involved in fertiliser transformations. In particular, due to nitrification, all ammonium-based treatments showed a bulk soil acidification of over 1.5 pH units and a relative increase in rhizo-soil pH as a consequence of nitrate uptake. The treatment with DMPP showed an opposite trend with a lower pH in rhizo-soil than in bulk soil. The ability of ammonium-fed plants to mobilise heavy metals from the non-labile pool was studied in another pot experiment using three soils with different properties and at different degree and type of heavy metal contamination. Whatever the soil, the metal concentrations in shoots were higher in plants fed with ammonium (ammonium sulphate plus DMPP treatment). Our results support the hypothesis that ammonium nutrition with nitrification inhibitors is a viable strategy to improve heavy metals phytoextraction while protecting bulk soil from acidification and presumably from metal leaching. An erratum to this article is available at .     

Identification of leukotriene D4 receptor binding sites in guinea pig lung homogenates using [3H]leukotriene D4

We have characterized [3H]leukotriene D4 binding to guinea pig lung homogenates. Both biphasic dissociation kinetics and curvilinear Scatchard plots indicated the presence of [3H]leukotriene high and low affinity states of the binding sites. The rank order of potency for the competition study was leukotriene C4 = leukotriene D4 greater than leukotriene E4 much greater than arachidonic acid, and for their contractile effect on lung strips was leukotriene C4 = leukotriene D4 = leukotriene E4 much greater than arachidonic acid. FPL-55712 was the only other agent tested that inhibited binding. These results suggest that binding of [3H]leukotriene D4 to the homogenate is consistent with its binding to specific leukotriene D4 receptor sites.     

Nicotinic Stimulation of [3H]Acetylcholine Release from Mouse Cerebral Cortical Synaptosomes

The effects of nicotine and 1,1-dimethyl-4-phenylpiperazinium (DMPP) on the release of newly synthesized [3H]acetylcholine in mouse cerebral cortical synaptosomes were examined. Nicotine and DMPP produced increases in [3H]acetylcholine release, over the level of spontaneous release, of 24% and 30%, respectively, of a maximum depolarization-induced release produced by 50 mM potassium. The maximum effect was achieved at a concentration of 1 X 10(-4) M for both agents. The time course of release indicated a slow onset of action, reaching a maximum effect at 15 min of incubation. Both nicotine and DMPP also produced a slightly greater release of total tritium, measured in the absence of cholinesterase inhibition, than of [3H]acetylcholine. The release induced by nicotine was completely antagonized by hexamethonium and was largely (58%) calcium-dependent. Nicotine also produced an increase in [3H]choline accumulation into synaptosomes. These results indicate that the nicotinic agonists nicotine and DMPP can produce a moderate enhancement of acetylcholine release by a receptor-mediated action on cholinergic nerve terminals in the central nervous system.     

Single-channel study of the binding-gating coupling in the slowly desensitizing chimeric α7-5HT3A receptor

We have studied the role of the highly conserved residue αLysine145 in the early steps of activation by acetylcholine of the nicotinic acetylcholine receptor (nAChR). Both macroscopic and single-channel currents were recorded in the slowly desensitizing chimeric mutant receptor α7V201-5HT3A/R432Q/R436D/R440A, made of α7 nAChRs and serotonin receptors of subtype 3A (ch1), and its corresponding mutant K145A (ch1/K145A) expressed in Xenopus oocytes. Mutant ch1/K145A receptors had a reduced gating function similar to that produced by the same mutation in the wild type receptor α7. The mutated receptor has reduced opening rate constants, β, and increased closing rate constants, α.