BACKGROUND: Bronchodilator treatment for asthma can be provided with various aerosolgenerating devices and methods. There have been no randomized trials of a breath-actuated nebulizer versus continuous 1-hour nebulization and/or small-volume constant-output nebulizer in pediatric asthma patients. METHODS: We conducted a randomized study of one-time albuterol treatment with the AeroEclipse breath-actuated nebulizer versus standard therapy (single treatment via small-volume nebulizer or 1-hour of continuous nebulized albuterol) in pediatric asthma patients in the emergency department. Eligible patients were those admitted to the emergency department, 0 months to 18 years of age, who presented with asthma or wheezing. We assessed all the patients with our clinical asthma scoring system and peak-flow measurement if possible. We stratified the patients by clinical asthma score and weight, and then randomized them to receive their initial albuterol treatment in the emergency department via either AeroEclipse or standard therapy. We recorded time in the emergency department, change in clinical asthma score, need for additional bronchodilator treatments, need for admission, patient response, ability to actuate the AeroEclipse, and adverse effects. RESULTS: We enrolled 149 patients between October 14, 2004 and November 11, 2005, and we randomized 84 patients to AeroEclipse and 65 to standard therapy. The cohort's average age was 5.5 years. There were no significant differences in demographics. The initial mean clinical asthma scores were 5.1 ؎ 2.4 in the AeroEclipse group, and 5.1 ؎ 2.1 in the standard-therapy group. Time in the emergency department was not different (AeroEclipse 102 min, standard therapy 125 min, P ‫؍‬ .10), but the AeroEclipse group had a significantly greater improvement in clinical asthma score (1.9 ؎ 1.2 vs 1.2 ؎ 1.4, P ‫؍‬ .001) and respiratory rate (P ‫؍‬ .002), and significantly lower admission rate (38% vs 57%, P ‫؍‬ .03). There was no difference in adverse effects. CONCLUSIONS: Although AeroEclipse did not reduce the time in the ED, it significantly improved clinical asthma score, decreased admissions, and decreased respiratory rate.

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Asparagine-linked glycosylation is an essential and highly conserved protein modification reaction. In eukaryotes, oligosaccharyl transferase (OT), a multisubunit membrane-associated enzyme complex, catalyzes this reaction in newly synthesized proteins. In Saccharomyces cerevisiae, OT consists of nine non-identical membrane proteins. Ost4p, the smallest subunit, bridges the catalytic subunit Stt3p with Ost3p. Mutation of transmembrane residues 18–24 in Ost4p has negative effect on OT activity, disrupts the Stt3p-Ost4p-Ost3p complex, results in temperature-sensitive phenotype, and hypoglycosylation. Heterologous expression and purification of integral membrane proteins (IMPs) are the bottleneck in membrane protein research. We report the cloning, successful overexpression and purification of recombinant Ost4p with a novel but simple method producing milligram quantities of pure protein. GB1 protein was found to be the most suitable tag for the large scale production of Ost4p. The cleavage of Ost4p conveniently leaves GB1 protein in solution eliminating further purification. The precipitated pure Ost4p is reconstituted in appropriate membrane mimetic. The recombinant protein is highly helical as indicated by the far-UV CD spectrum. The well-dispersed HSQC spectrum indicates that this minimembrane protein is well folded. The successful production of pure recombinant Ost4p with a novel yet simple method may have important ramification for the production of other membrane proteins.

Glutaminase interacting protein (GIP), a PDZ domain containing protein, mediates the distribution and clustering of proteins/peptides in membranes, acting as a scaffold where signaling molecules are linked to a multi-protein complex. GIP has been shown to play a key role in the glutamate signaling system. Some metals, particularly Pb2+, Cu2+and Zn2+, have been implicated in a wide range of neurological disorders including Alzheimer's disease and Parkinson's disease, whose etiologies have been associated with dysfunction of the glutamate signaling system. Here, for the first time, the effects of lead, copper, and zinc on GIP were determined by using circular dichroism and fluorescence spectroscopy. All three metal ions significantly affected the conformational properties of GIP. The deconvolution of CD data showed that, with increasing amounts of Pb2+/Cu2+/Zn2+, theα-helix percentage decreased while theβ-sheet content increased. The binding constants of GIP to Pb2+, Cu2+and Zn2+determined by fluorescence were found to be 1.4, 2.38 and 2.85 μM respectively, which indicated strong bindings between GIP and all three metal ions. We propose that the metal ion binding site of GIP is located onα-2 helix, where residues His90, Asp91 and Arg94 may coordinate the metal ions. The conformational change of GIP upon metal ion binding possibly results from the disruption of a salt bridge between Asp91 and Arg94.