A gas chromatography method, coupled with diode array photometric spectral detection in the ultraviolet region (167-330 nm), was developed for the analysis of the gas phase of cigarette smoke. The method enabled us to identify more than 20 volatiles present in the vapor phase of cigarette smoke. In that way, all major volatile organic compounds (including aldehydes, conjugated dienes, ketones, sulfides, furans, and single-ring aromatics), as well as nitric oxide (NO) and hydrogen sulfide (H(2)S), can be analyzed in a straightforward manner through a single chromatographic run of <50-min duration. The method can easily be applied by the introduction of a small volume of the gas-phase stream into the GC injection loop directly through the smoking apparatus exhaust circuit, thus providing an excellent alternative to available methods, which usually require extraction or concentration steps prior to any chromatographic analysis. Furthermore, all problems concerning aging of the gas phase are eliminated. Twelve compounds (including NO) were chosen for quantification through the use of appropriate calibration standards. Comparison of the vapor phase yields of these compounds for the reference cigarette Kentucky 1R4F with already reported data indicates that this method is very reliable as far as accuracy and reproducibility of the results are concerned. Finally, the proposed methodology was used to compare the concentration of these cigarette smoke gas-phase constituents among individual puffs.

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BackgroundCigarette smoke (CS) is the main cause in the development of chronic obstructive pulmonary disease (COPD), the pathogenesis of which is related to an extended inflammatory response. In this study, we investigated the effect of low and high doses of gas phase cigarette smoke (GPS) on cultured lymphocyte progenitor cells, using techniques to assess cell viability and to elucidate whether cells die of apoptosis or necrosis upon exposure to different doses of GPS.MethodsIn our approach we utilised a newly-established system of exposure of cells to GPS that is highly controlled, accurately reproducible and simulates CS dosage and kinetics that take place in the smokers' lung. This system was used to study the mode of cell death upon exposure to GPS in conjunction with a range of techniques widely used for cell death studies such as Annexin V staining, activation of caspase -3, cytoplasmic release of cytochrome C, loss of mitochondrial membrane potential and DNA fragmentation.ResultsLow doses of GPS induced specific apoptotic indexes in CCRF-CEM cells. Specifically, cytochrome C release and cleaved caspase-3 were detected by immunofluorescence, upon treatment with 1-3 puffs GPS. At 4 h post-exposure, caspase-3 activation was observed in western blot analysis, showing a decreasing pattern as GPS doses increased. Concomitant with this behaviour, a dose-dependent change in Δψm depolarization was monitored by flow cytometry 2 h post-exposure, while at 4 h Δψm collapse was observed at the higher doses, indicative of a shift to a necrotic demise. A reduction in DNA fragmentation events produced by 5 puffs GPS as compared to those provoked by 3 puffs GPS, also pointed towards a necrotic response at the higher dose of GPS.ConclusionCollectively, our results support that at low doses gas phase cigarette smoke induces apoptosis in cultured T-lymphocytes, whereas at high doses GPS leads to necrotic death, by-passing the characteristic stage of caspase-3 activation and, thus, the apoptotic route.

In line with recent developments in leadership research which conceptualise leadership as a discursive and collaborative process rather than a set of static attributes and characteristics displayed by individuals, this paper explores some of the discursive processes through which leadership emerges in a sports team. Drawing on over ten hours of naturally occurring interactions among the players of a women’s netball team in the UK, and applying the concepts of deontic and epistemic status and stance, we identify and describe some of the specific processes through which leadership is claimed and assigned, as well as rejected, passed on, and eventually accepted by different team members at different points throughout an interaction. While the processes outlined in our analysis contribute to theoretical discussions regarding the notion of emergent leadership, this paper also demonstrates the benefits of taking a discourse analytical approach to leadership, and outlines how such an approach enables researchers to empirically capture emergent leadership in situ.