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After 24 hours, supernatants were harvested and again analysed for the expression of CXCL8 and TNF-α.

DNA from corresponding normal colon from this patient was again analysed for the presence of this mutation.

Afterwards, the platelets were centrifuged at 1900× g for 10 min and the supernatant was frozen at −80 °C, while the pellet was dissolved in 100 µl of 0.4 N NaOH (1 h, 45 °C) and again analysed for total protein content.

Then this resulting reaction mixture was irradiated with UVA for 1 h and again analysed by HPLC-ESI-MS.

These will then be analysed for signs of biological activity.

Each one of the 146 patients was again retrospectively analysed using a single follow-up BS for measurement of BSI values during ADT.

Mutants in which the plasmid insertion was in a coding region were selected, re-transfected with the recovered knockout plasmid (Fig. S1) and analysed again for defective growth.

Therefore, to eliminate any doubt over allele size scoring due to technical modifications, the entire collection of genotypes was analysed again for this study using exactly the same equipment and fluorochrome primers.

Again, separate analyses for each professional group were conducted.

After being stained for surface molecules, the cells were washed again and analysed with a FACSCalibur flow cytometer (BD Biosciences).

Finally, the effect of smoking status on direct, indirect and total healthcare costs was analysed, again adjusting for age, sex, education, alcohol consumption and physical activity.