SUN Tirol: Smoking undone naturally in Tirol

Effectiveness of the biofeedback method to sustain smoking cessation measured by comparing the number of ‘smoke-free’ participants in the groups with and without biofeedback until at least the end of the study

1. Achievement of the planned number of participants (n=200) in Tirol within 5 months measured using study records of recruitment at 6 months after recruitment initiation
2. Number of participants in the two groups (with and without biofeedback) who have completed the study (regardless of whether they continue smoking) measured using study records from baseline to 6 months of recruitment time
3. Epigenetic alterations as a result of smoking cessation evaluated using DNA methylation signatures associated with age, disease, or exposure, e.g. WID indices (WID-BC, WID-OC, WID-SMKand WID-age), epigenetic clocks, etc. and measured using microarray analysis (Illumina HumanMethylationEPIC) using of buccal and cervical/vaginal smears and blood at baseline and 2, 4, 6 and 8 months after study entry. Per-protocol and intention to treat analyses will be conducted and compared between the two groups at respective timepoints, and separately comparing changes from baseline in each group.
4. Clinical factors, including resting heart rate and blood pressure (measured using the device Boso Medicus X (Bosch + Sohn)) and vascular elasticity (measured using the device Vicorder®) at baseline and 2, 4, 6 and 8 months after study entry. Per-protocol and intention to treat analyses will be conducted and compared between the two groups at respective timepoints, and separately comparing changes from baseline in each group.
5. Quality of life (measured using the WHO quality of life questionnaire (WHOQOL)), quality and duration of sleep (measured using the Pittsburgh Sleep Quality Index (PSQI)) and stress level (measured using the Perceived Stress Scale (PSS-10)) at baseline and 2, 4, 6 and 8 months after study entry. Per-protocol and intention to treat analyses will be conducted and compared between the two groups at respective timepoints, and separately comparing changes from baseline in each group.
6. Blood values, including:
6.1. Blood count and differential blood count measured using flow cytometry on a Sysmex XN-3000 module at baseline and at the end of the study (eight months after study entry). All laboratory parameters are measured using CE-IVD test systems and ISO9001:2015 procedures. Per-protocol and intention to treat analyses will be conducted and compared between the two groups at respective timepoints, and separately comparing changes from baseline in each group.
6.2. LDL, HDL, HbA1c, creatinine and CRP measured using CE-IVD test systems and ISO9001:2015 procedures at baseline and at the end of the study (eight months after study entry): CRP, HDL and LDL are measured by immunologic methods (CLIA) using a Roche Cobas 801 module. HbA1c is measured by an immunologic method (TINIA) using a Roche Cobas 513 analyzer and Creatinine is measured by an enzymatic method using a Roche Cobas c702 analyzer. Per-protocol and intention to treat analyses will be conducted and compared between the two groups at respective timepoints, and separately comparing changes from baseline in each group.
6.3. Analysis of cellular measurements: distribution of the different immune cell populations (% and absolute values) in peripheral blood mononuclear cells (PBMCs); activation levels of monocytes and T-cells and expression of adhesion molecules measured using RT-qPCR, cell culture experiments, or flow cytometry. Per-protocol and intention to treat analyses will be conducted and compared between the two groups at respective timepoints, and separately comparing changes from baseline in each group.
6.4. Inflammatory cytokines and factors and blood (plasma), including IL-1, HMGB-1, RAGE, sASC, and IL-6 measured using enzyme-linked immunosorbent assays (ELISA) or RT-qPCR compared between the two groups at respective timepoints 2, 4, 6, and 8 months after study including and separately comparing changes from baseline in each group.
6.5. Cytotoxicity of peripheral blood mononuclear cells measured using e.g. the xCELLigence assay between the two groups at respective timepoints 2, 4, 6, and 8 months after the study including, and separately comparing change from baseline in each group.