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5, Messer Griesheim, Frankfurt, Germany) with a known ��13C value of ?23.64�� (V-PDB) was used as the working reference gas. All ��13C values were normalized relative to V-PDB using a CH4 standard. Samples were routinely analysed three times (n?=?3) and the average standard deviations of the GC/C/IRMS measurements were in the range of 0.1 to 0.3��. To evaluate the effect of NaN3 treatments on CH4 production (Fig.?2) and Ibrutinib in vivo the effect of various inhibitors (Fig.?5) one-way analysis of variance (anova) and a Tukey- Honestly Significant Difference (HSD) analysis were employed. Levels of significances were defined as follows: P?MMP23B were carried out with the software package SPSS version 17 (Chicago, IL, USA). In an initial set of experiments, tobacco cells (Nicotiana tabacum cv. BY-2) were studied as these cells are often used as a model system in molecular cell biology (Nagata, Sakamoto & Shimizu 2004). We monitored the change of CH4 concentration in the headspace of the incubation vials containing tobacco BY-2 cells when incubation under sterile conditions in the dark at 26?��?1?��C was carried out for 19?h (Fig.?2a). The untreated cell cultures (control) showed minor CH4 uptake rates [?6.4?��?2.6?ng?CH4?g?1 dry weight (DW)], whereas treatment with NaN3 showed substantial formation of CH4 at all concentrations tested (P?learn more explanation for this could be that at the higher NaN3 concentrations faster cell mortality interrupts transport of the precursor compounds to the site of CH4 formation. In order to determine if azide-induced CH4 emission from plant cells is a general phenomenon two other plant species were examined: cells of grape vine (Vitis vinifera) and sugar beet (Beta vulgaris L.). The emission pattern for sugar beet cell cultures is shown in Fig.?2b, where it can be seen that there was slight, but not significant, CH4 uptake for the control samples (?2.9?��?2.6?ng?g?1 DW), whereas for the 1 and 5?mM NaN3 treatments CH4 emissions were similar (3.5?��?2.2 and 5.6?��?1.2?ng?CH4?g?1 DW, respectively). CH4 emissions from cells treated at the 10?mM level were highest at 16.8?��?13.8?ng?g?1 DW, which is highly significant formation (P?