1. ICEmB Capri, September 29 - October 2 2002 THz exposure of whole blood for the study of biological effects on human lymphocytes M. R. Scarfì, M. Romanò, R. Di Pietro, O. Zeni CNR-IREA Napoli (I) A.Doria, G. P. Gallerano, E. Giovenale, G. Messina, A. Lai, G. Campurra, D. Coniglio, M. D’Arienzo ENEA – FIS-ACC Frascati (I)

2. ICEmB Capri, September 29 - October 2 2002 To study the potential health hazard of THz radiation since very little is known about its interaction with biological systems Specialised personnel and users in the laboratory-scale THz sources in use at research labs technical personnel and future patient exposure due to recent technological applications of THz radiation in biology and medicine, based on the specific spectroscopic fingerprints of biological matter in this spectral region

3. ICEmB Capri, September 29 - October 2, 2002 Genotoxic effects a close correlation exists between DNA damage and cancer occurence Human peripheral blood lymphocytes well known biological model Cytogenetic techniques cytokinesis block MN assay is a very sensitive and simple indicator of chromosome damage, which also provides information on cell cycle progression AIM: to study MN induction in HPBL exposed in G 0 phase to 120 and 130 GHz THz radiation

4. ICEmB Capri, September 29 - October 2, 2002 THz Irradiation Compact FEL Facility at the ENEA Research Center a b c d a = light pipe b = TDS c = right-angle bend d = pyroelectric detector

5. ICEmB Capri, September 29 - October 2, 2002 Absorption measurements on whole blood at 120 GHz Optimal thickness of the sample: 1 mm Optimal irradiation geometry to contain a minimum useful whole blood volume of 2 ml: Petri dishes with internal diameter of 52 mm

6. ICEmB Capri, September 29 - October 2, 2002 fBunch durat. Bunch interv. Pulse durat. Pulse rep. rate Aver. power Irrad. time Delivered energy 120 GHz 50 ps330 ps4 µs2 Hz1 mW20 min. 1.20 J 130 GHz 50 ps330 ps4 µs2 Hz0.6 mW 20 min. 0.72 J Irradiation Parameters

7. ICEmB Capri, September 29 - October 2, 2002 Blood collection at the ENEA occupational health unit by venipuncture in heparinized vacutainers Donors 9 healthy subjects aged between 30 and 45 years (mean age: 39.33 ± 4.82 y) neither therapeutic drugs nor diagnostic X rays

8. ICEmB Capri, September 29 - October 2, 2002 Biological procedure for each donor, 3 conditions: exposed sham exposed control 2 ml wb in 52 mm Petri dishes Sham exposure rationale: to simulate the same environmental conditions of exposed samples, but in absence of radiation

9. ICEmB Capri, September 29 - October 2, 2002 Lymphocyte cultures Following 20 min. exposure/sham exposure, 0.8 ml wb in 9.2 ml RPMI 1640 medium containing 15% FCS, 2 mM L-Glutamine and 100 µl PHA control cultures handled in parallel

10. ICEmB Capri, September 29 - October 2, 2002 Cytokinesis-block micronucleus assay Screening MN frequency: % CB cells containing MN/1000 CB cells CBPI: by classifying 500 cells according to the n° of nuclei [M1 + 2M2 + 3(M3+M4)/N] Statistical analysis two tailed paired Student’s t test

11. ICEmB Capri, September 29 - October 2, 2002 1000X

12. ICEmB Capri, September 29 - October 2, 2002 MN frequency in HPBL cultures following THz radiation exposure in G 0 phase (9 subjects) at 120 and 130 GHz P>0.05 120 GHz – 6 donors 130 GHz – 3 donors

13. ICEmB Capri, September 29 - October 2, 2002 CBPI in HPBL cultures following THz radiation exposure in G0 phase (9 subjects) at 120 and 130 GHz P>0.05

14. ICEmB Capri, September 29 - October 2, 2002 Conclusions Environmental condition at FEL laboratory does not influence MN induction and cell cycle kinetics of HPBL (sham/control); THz exposure of HPBL, at both 120 and 130 GHz: does not affect MN formation; does not influence cell proliferation (sham/exposed). Ongoing studies on a larger number of individuals should provide further, more complete information on this topic.

15. ICEmB Capri, September 29 - October 2, 2002 Future work exposed/sham exposed different exposure conditions: f (following input from WP1); power levels modulation conditions: still open possibility of extending the irradiation measurements to the range 1-10 THz at other partners sites

16. ICEmB Capri, September 29 - October 2, 2002