Chemiluminescence measurement of TNFα, TGF-β and IFN-γ serum concentrations.

Frequencies and severity of infectious diseases will be evaluated by modified Infectious Diseases Questionnaire (GNC).

This questionnaire includes questions on infectious diseases of upper and lower respiratory tract, gastrointestinal tract, skin and urogenital tract contracted during the previous 12 months. Questions investigate on the number and duration of infections, necessity of antibiotic or antifungal therapy, hospital stay and days of absence from work. Final score represents the frequency of infections. Moreover, some questions investigate possible susceptible or protective factors for infectious diseases: vaccinations, use of corticosteroids, concomitant diseases, previous appendectomy, tonsillectomy, adenoidectomy, splenectomy or thymectomy.

Composite outcome consisting of simultaneous measurement of calcium channels and related proteins: Cav1, Calcium Release Activating Channel, Stromal Interaction Molecule 1 (STIM1), Calcium release-activated calcium modulator 1 (encoded by Orai1 gene), Inositol trisphosphate receptor, Nuclear factor of activated T-cells, Calcineurin and Nuclear Factor Kappa-Light-Chain-Enhancer of Activated B cells (NF-kB) expression evaluated by Prime Polymerase chain reaction

Quality of life will be evaluated by the Physical Component score and the Mental Component score of the self-administered questionnaire SF-36-Item Health Survey questionnaire.

This questionnaire measures eight scales: physical functioning, role physical, bodily pain, general health (physical component) and vitality, social functioning, role emotional, mental health (mental component).

Interpretation of the score will be the following: at each item of the questionnaire corresponds a percentage value (from 0% to 100%). The average of the single items constitutes the scale total percentage (from 0% to 100%); missing data are not considered during calculation. High score defines a more favorable health state.

Psychometric evaluation will be assessed by the scores of two self-administered questionnaires: Beck depression Index and Middlesex Hospital Questionnaire.

Beck depression Index is a 21-item measure of depressive symptoms. Each answer is scored on a scale value (from 0 to 3 points). The global score is obtained adding all single scores, with a range of 0-63. Higher score constitutes worse burden of symptoms (0-13: minimal depression, 14-19: mild depression, 20-28: moderate depression, 29-63: severe depression) Middlesex Hospital Questionnaire is a self-rating inventory measuring aspects of six distinct categories of psychoneurosis and affective status. Considering symptoms gravity or frequency each answer is scored on a scale value (from 0 to 2 points). Category scores derive from the addition of the scores of 8 questions. Specific cut off are used to distinguish between normal and pathological behavior (anxiety 5, phobia 5, obsession 7, somatization 5, depression 4 and hysteria 4).

Sleep disturbances will be evaluated by The Pittsburgh Sleep Quality Index (PSQI). This questionnaire contains 19 self-related questions which are combined to create 7 component scores with a range of 0-3 points (0:no difficulty, 3: severe difficulty). Global score is the result to the addition of all component scores with a range of 0-21 points (0:no difficulty, 21: severe difficulty).