Real-time breath monitoring of propofol and its volatile metabolites during surgery using a novel mass spectrometric technique: a feasibility study.

BACKGROUND: At present, there is no rapid method for determining the plasma concentration of i.v. anaesthetics. A solution might be the measurement of the anaesthetic concentration in expired breath and its relation to the plasma concentration. We used chemical ionization methods to determine whether an i.v. anaesthetic can be detected in the low concentrations (parts per billion by volume) in the expired breath of an anaesthetized patient. METHOD: Chemical ionization mass spectrometry can measure trace gases in air with high sensitivity without interference from major gases. We carried out a feasibility trial with a proton transfer reaction mass spectrometer (PTR-MS) to monitor the i.v. anaesthetic agent propofol and two of its metabolites in exhaled gas from an anaesthetic circuit. Exhaled gas was sampled via a 4 m long, unheated tube connected to the PTR-MS. RESULTS: Propofol and its metabolites were monitored in real time in the expired breath of patients undergoing surgery. CONCLUSION: Routine measurement of i.v. agents, analogous to that for volatile anaesthetic agents, may be possible.

Direct measurement of a pure rotation transition in H2(+).

The N = 1<--0 pure rotation transition in the nu = 19 level of the ground electronic state of H2(+) was observed at 14,961.7+/-1.1 MHz. Recent theory predicts significant electric dipole intensity in forbidden rotation and rotation-vibration transitions involving levels near the dissociation limit; the relevant levels are bound by only 0.74 and 0.22 cm(-1). The transition was predicted to have a transition moment of 0.42 D; our measurement is consistent with this value.

Probing the details of the HIV-1 Rev-Rev-responsive element interaction: effects of modified nucleotides on protein affinity and conformational changes during complex formation.

The solution structure of the human immunodeficiency virus type 1 (HIV-1) Rev-responsive element (RRE) has been investigated by enzymic and chemical structural probing of a 71 nt RRE transcript. The minimum sequence information required to maintain recognition by the Rev protein has previously been mapped to a 29 nt stem-loop structure, known as minSLIIB. The key recognition target is a single-stranded RNA bubble at the base of the RNA stem. The fine details of RNA recognition have been probed using chemically synthesized minSLIIBs containing variant base or sugar residues at sites within the bubble. These have been analysed by gel retardation assays and their relative affinities for Rev protein determined. Complex formation between the wild-type minSLIIB RRE and Rev protein was also monitored using CD spectroscopy, which suggests a change in RNA conformation upon Rev binding. The spectral change is consistent with localized melting of RNA, leading to a decrease in the level of base stacking and/or a change in base tilting, during formation of the complex. Deoxynucleotide substitutions on just one side, the 5' side, of the bubble inhibit the conformational change detected by CD. The data are consistent with a dynamic interaction between Rev and its target site. The contact points between Rev and the RRE were probed directly using photo-cross-linking with either ribo-5-bromouridine- or ribo-4-thiouridine-substituted minSLIIBs. The data are consistent with protein-RNA contacts at the bottom of the bubble.

Modeling and solution structure probing of the HIV-1 TAR stem-loop.

We present a model for the three-dimensional structure of the HIV TAR stem-loop, based on a modeling algorithm which makes use of the known X-ray coordinates of tRNAs to generate a model structure, which has then been tested experimentally in solution by enzymatic and chemical structure probing of ribo-oligonucleotides encompassing the TAR sequence. The modeling suggested that the structure of TAR was similar to that of the anti-codon loop of tRNA(Asp), having a loop of just three single-stranded residues with a mismatched adenine excluded from the helical stem on the 3' side of the loop. The structural probing is consistent with such a structure for the loop, and reveals an unusual structure around the 5' uridine-rich bulge, which is the binding target for the transactivator protein Tat. These data may be useful in understanding the interaction of TAR with the Tat protein and may aid in the design of anti-AIDS drugs. The coordinates of the model are available on request.