icemb
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ICEmB. 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). - PowerPoint PPT PresentationTRANSCRIPT
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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’ArienzoENEA – FIS-ACC Frascati (I)
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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
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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 G0 phase to 120 and 130 GHz THz radiation
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ICEmB
Capri, September 29 - October 2, 2002
THz Irradiation Compact FEL Facility at the ENEA Research Center
a
b
cd
a = light pipeb = TDSc = right-angle bendd = pyroelectric detector
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ICEmB
Capri, September 29 - October 2, 2002
Absorption measurements on whole blood at 120 GHz
Optimal thickness of the sample: 1 mmOptimal irradiation geometry to contain a minimum useful whole blood volume of 2 ml: Petri dishes with internal diameter of
52 mm
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ICEmB
Capri, September 29 - October 2, 2002
f Bunch durat.
Bunchinterv.
Pulse durat.
Pulse rep. rate
Aver.power
Irrad. time
Delivered energy
120 GHz
50 ps 330 ps
4 µs 2 Hz 1 mW 20 min.
1.20 J
130 GHz
50 ps 330 ps
4 µs 2 Hz 0.6 mW
20 min.
0.72 J
Irradiation Parameters
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ICEmB
Capri, September 29 - October 2, 2002
Blood collectionat the ENEA occupational health unitby venipuncture in heparinized vacutainers
Donors9 healthy subjects aged between 30 and 45 years (mean age: 39.33 ± 4.82 y)
neither therapeutic drugs nor diagnostic X rays
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ICEmB
Capri, September 29 - October 2, 2002
Biological procedurefor 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
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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
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ICEmB
Capri, September 29 - October 2, 2002
Cytokinesis-block micronucleus assay
Screening MN frequency: % CB cells containing MN/1000 CB cellsCBPI: by classifying 500 cells according to the n° of nuclei
[M1 + 2M2 + 3(M3+M4)/N]
Statistical analysistwo tailed paired Student’s t test
t=0 t=44h t=72h x - - - - - - - - - - - - - - - - - - - - x- - - - - - - - - - - - - - x | | | Start cyt- B harvest and (PHA) slide preparation
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ICEmB
Capri, September 29 - October 2, 2002
1000X
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ICEmB
Capri, September 29 - October 2, 2002
MN frequency in HPBL cultures following THz radiation exposure in G0 phase (9
subjects)at 120 and 130 GHz
00,20,40,60,8
1
120 GHz 130 GHz
Exposure frequency
MN
freq
uenc
y
controlshamexposed
P>0.05
120 GHz – 6 donors130 GHz – 3 donors
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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
00,5
11,5
22,5
120 GHz 130 GHz
Exposure frequency
CB
PI
controlshamexposed
P>0.05
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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.
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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