The Laboratory of Radiation...
Transcript of The Laboratory of Radiation...
The Laboratory
of Radiation Biology
Page 2
Andrej Vladimirovich
Lebedinskij
Vasilij Vasil'evich Parin Oleg Georgievich Gazenko
Jurij Grigor'evich Grigor'ev
Founders
Page 3
First radiobiological experiments on synchrocyclotron
Page 4
History
1959 First experiments at Laboratory of Nuclear Problems (LNP)
1978 Biological Research Sector at LNP
1988 Biological Department at LNP
1995 The Department of Radiation and Radiobiological Research
2005 Laboratory of Radiation Biologywww.lrb.jinr.ru
Prof. E.A. Krasavin,
Corr.Member of RAS
Page 5
The LRB performs education programs for students and young researchers
on modern equipment
Education activity
Page 6
Main research topics
1. Research on the effect of accelerated heavy ions of different
energies on genetic structures
2. Research on the effect of different doses of accelerated charged
particles on the retina, study of cataractogenesis
3. Research on the character of the heavy charged particle-induced
damage and functional disorders of central nervous system (CNS)
cells.
4. Mathematical modeling of radiation induced effects in
biophysical systems
5. Evaluation of the radiation environment and radiation safety
6. Solving problems of astrobiology (in cooperation with Italy)
Page 7
JINR’s accelerators
Phasotron: protons 660 MeV
U-400: heavy ions 10 MeV/u
U-400M: heavy ions 50 MeV/u
Nuclotron: heavy ions up to 4 GeV/u
Page 8
1 unit of the dose 1 unit of the dose
X-rays Fe ion
The dose distribution of radiation in matter
Page 9
The Galactic Cosmic Ray (GCR) energy spectrum
The integral flux of GCR particles of
carbon and iron groups equals to 105
particles/cm2 per year
Particle flux density
interplanetary space Z20
160 per day per cm2
Page 10
Consequences of Galactic heavy ion action
Formation of gene and structural
mutations;
Induction of cancer;
Violation of visual functions:
lesions of retina;
cataract induction;
CNS violation
DNA damage
Page 12
Single strand break
Base damage
Sugar damage
Isolated DNA damage
Page 13
Clustered DNA damage
Double strand break
Sugar damage
Base damage
Base damage
Page 14
D = 0 Gy D = 5 Gy
D = 40 GyD = 20 Gy
D =10 Gy
D = 60 Gy
“Comet assay” for detection of DNA lesions
Dose, Gy
mt
Page 15
Visualization of damaged sites in DNA
3D analysis of induced
γH2AX/53BP1 foci
- Acquiarium
Irradiation
Fixation of cells at different times
post-irradiation (PI)
Visualisation of induced DSBs
(γH2AX/53BP1 foci)
Acquisition of images
H2AX
DSB DNA
H2AX
Primary antibody
Secondary antibody
with fluorescent dye
53ВР1 53ВР1
γH2AX foci53BP1 foci
merge
Page 16
Human cells exposed to γ-rays and 11B ions
5 min
24 h
1 h
γ-rays (LET = 0.3 keV/μm) 11B ions (LET = 135 keV/μm)
x-y
2 μm
x-y
x-y
x-y
x-y
x-y
y-z
y-z
y-z
y-z
y-z
y-z
x-z
x-z
x-zx-z
x-z
x-z
γH2AX 53BP1 Chromatin (DAPI)
Page 17
Human cells exposed to 11B ions at 10°
4 h
30 min15 min5 min
1 h 24 h
γH2AX 53BP1 Chromatin (DAPI)
Page 18
Kinetics of the formation and disappearance ofγH2AX/53BP1 foci
0
20
40
60
80
Ave
rage
nu
mb
ero
fγH
2A
X/5
3B
P1
fo
cip
er c
ell
Time after irradiation
Comparison of γH2AX/53BP1 foci:
γ-rays and 11B
γ-rays
11Bor
γ-rays
11Boron
Page 19
Incidence of clusters of γH2AX/53BP1 foci
0
10
20
Ave
rage
nu
mb
er
of
clu
ste
rs p
er
cell
Time after irradiation
Comparison of total clusters:
γ-rays and 11B
11Borγ-rays
Page 20
Monte-Carlo computer modeling of heavy ion tracks and cluster damage analysis
GEANT4-DNA
http://geant4.org
Mutagenesis and RBE
Page 22
Guanine
Gene mutation Structural mutation
Radiation induced mutagenesis
Page 23
Dose, Gy
Nm
/N 1
0-5
Dose, Gy
4He 50
4He20
12C200
Nm
/N
а б
The frequency of gene and structural mutation
induction after γ-ray and heavy ion irradiation
Page 24
luciferase
FMNH2+ RCHO + O2 FMN + RCOOH +H2O + h
Induction of mutagenic DNA repair by heavy ions
Page 25
0,8
1,0
1,2
1,4
1,6
1,8
2,0
2,2
2,4
2,6
2,8
3,0
0,1 1 10 100 1000
x
x
x
x
L E T , keV/m
R B
E
x
3,2
1
2
3
RBE dependence on LET
1 – gene mutations
2 – deletions
3 – lethal effect
Page 26
Formation of unstable chromosomal aberration in
human cells after heavy ion irradiation
Unstable
chromosomal
aberration
Block of cell division
Ab
erra
tio
ns/
10
0 c
ells
Dose, Gy
Page 27
Stable
chromosomal
aberration
Formation of stable chromosomal aberrations in
human cells after heavy ion irradiation
Successful of cell division
Tra
nsl
oca
tio
ns/
10
0 c
ells
Dose, Gy
Chromosome № 1
Page 28
Cytogenetic effect of low doses of accelerated 24Mg ions
The frequency of cells with
chromosome aberrations.
Chinese hamster cells exposed to 24Mg
ions with energy 500 MeV/nucleon
micronuclei
chromosomal aberrations
24Mg
γ-rays
Page 29
Genetic network model of induced mutagenesis in bacteria E.coli
Page 30
Mathematical modeling of DNA repair systems in bacteria
Page 31
Mathematical modeling of DNA repair systems
in mammals and human
Action of radioprotectors
Page 33
Influence of radioprotectors on bacterial cells
after heavy ion irradiation
bacteria E.coli
without protector
cysteamine
LET, keV/mkm
Cancer
Page 35
Gardner tumors
H a r d e r ia n G la n d T u m o r P r e v a le n c e
D o s e , G y
0 .0 0 .5 1 .0 1 .5 2 .0
Re
lativ
e R
isk
1 0
2 0
3 0
G a m m a
p r o to n
h e l iu m
n e o n
ir o n ( 6 0 0 M e V /u )
i r o n ( 3 5 0 M e V /u )
-rays
Iron ions
Nelson, 2006
Page 36
Skin cancer (rats)
Burns, Albert, 1986
Page 37
RBE for carcinogenic effect of irradiation
Eyes and retina
Page 39
Eye lens and retina UV-induced aggregation of
L- crystalline under B11
cytoplasm micro-vacuolization, fiber
cell swelling, nuclear fragmentation
Cataractogenesis
А Control Б МНМ, 70 mg/kg, 2h
Dysfunction after mutagen
insertion
electroretinogram
Ostrovskii М.А., 2011
Page 40
Worgul et al., 2006
Dose, Gy Dose, Gy
1000
0.01 0.1 1 10
Cata
ract
ratio
0.1
1
10
Iron ions
X-rays
RBE = Dx/DFe
0.01 0.1 1R
BE
1
10
100
Iron ions
Cataract induction by iron ions and X-rays
Page 41
20000
30000
40000
50000
60000
70000
80000
90000
Arb
itra
ry u
nit
s
0 50 100 150 200 250
*
dose, cGy
50 cGy
0 cGy
200 cGy
10 cGy
100 cGy
5 cGy
Action of 56Fe ions on retina cells
Vazquez, 2006
Axon growth index vs 56Fe ion dose
Apoptosis
Central Nervous System
A B
A B
Fe
p
p
p
e
Mixed Field
Multi-hit
Cosmic ray hit frequencies in CNS critical areas
CNS in General
2 or 13% cells will be hit at least one Fe
particle
8 or 46% would be hit by at least one
particle with Z15
Every nucleus will be traversed by a
proton once every 3 days and a alpha
particle once every 30 days.
0 cGy 50 cGy
100 cGy 200
cGy
TRACK DIRECTION
FE ION TRACKS VISUALIZED BY MARKERS OF DNA DSBs (γH2AX)
Page 44
Cognitive tests (Morris water maze)
1 month after
irradiation
M.Rabin, 2005
Page 45
Persistent reduction in the spatial learning ability of rats
after 56Fe ion irradiation
0
20
40
60
80
100
120
140
160
1 2 3
Симуляция облучения 20 сГр 1 ГэВ/нуклон Fe
day of test
Del
ay, s
Results after 3 months
Ф 105/cm2
20 cGy
R. Britten et al., 2012 20 cGy 1GeV/nucleon 56Fecontrol simulation
Page 46
First experiments with monkeys
Irradiation with a proton
medical beam, 170 MeV
Irradiation with 12C
ions, 500 MeV/u, at the
Nuclotron
Page 47
Distinguishing visual stimuli on the sensory attributes
conditioned stimulus
color
brightness
configuration
right response
refreshment
orientation
Page 48
Molecular targets at subcelular level
RRP in glutamate
synapses
Levels of receptor
subunits
Shi et al (2006)
Machida et al (2010)
Britten et al (2014)
Myelin sheath degradation
Chiang et al (1993)
Na+ currents
Mullin et al (1986); Hunt et al (1988);
Sokolova et al (2015)
Membrane
hyperpolarization
Sokolova et al (2015)
Page 49
Rarefaction of Purkinje cell layer after irradiation
by 645 МeV protons and 137Cs -rays
Krasavin Е.А., 1979
Page 50
Irradiation with 1 Gy of 500 MeV/u carbon ions
Radiation-induced decrease in the level of neurotransmitters is observed
in the brain regions responsible for the emotional and motivational state
Studying the level of neurotransmitters in different rat
brain areas
3 months after irradiation
Prefrontal
cortex
Nucleus
accumbens
Rat
brain
HippocampusStriatum
Hypothalamus
Page 51
Modeling of energy deposition events in CA1 pyramidal neurons
Page 52
Modeling of cognitive tests by neural networks
Simulation of single neuron and
network activity during WM task
40cGy, Fe ions20cGy
Page 53
The used risk concept
Based on the introduction of a generalized dosimetric functional as the
criterion and quantitative measure of radiation danger
Generalized dose HI and HD for the evaluation of, respectively, the
immediate adverse consequences during the flight and the delayed
consequences during the rest of life:
HI = ( Di × KKIi × KBIi × KРIi) KMIΣi = 1
n __
ККi – radiation quality coefficients;
КВi – coefficient taking into account the dose distribution over time;
КРi – coefficient taking into account the dose distribution over the human body;
КМ – coefficients of the organism’s radiation response modification caused by
other factors of the space flight.
HD = ( Di × KKDi × KBDi × KРDi) KMDΣi = 1
n __
Page 54
Prad. damage = PСNS + Рimmed. eff. + Pdelayed eff. + Pother
The probability of successful mission implementation
P = 1 – (Prad. damage + Pnon-rad. injury + Ptechn. failure)
Page 55
Astrobiology
Page 56
Formamide as Prebiotic Probe
“Simulations based on Density Functional Theory show that formamide is the most stable species with molecular formula “CHON”Pauzat, F. et al. 6th EANA 2006
“This one-carbon molecule was detected in the gas phase of interstellar medium” Crovisier, J. Astrobiology: Future Perspectives, Kluwer Eds, 2004 Chapter 8, p. 179-203.
“in the long period comet Hale-Bopp“ Bockelee-Morvan,D. et al. Astr. Astrophys. 2000
“in the solid phase on grains around the young stellar object W33A “W.A. Schutte et al. Astr. Astrophys. 1999
Jupiter's satellite Europa
«in the gas surrounding IRAS 16293-2422, a sun-like forming star in the Rho Ophiuci nebula». Astrophysical Journal Letter (ApJ 763, L38), 2013
JINR-Cyclotron
In each tube:
FORMAMIDE
ALONE
RADIATION
FORMAMIDE
+ METEORITE
RADIATION
FORMAMIDE
+ METEORITE
Absence of
RADIATIONNO PRODUCTS
Few amounts
Formamide irradiation at U400M cyclotronMeteorites: Dohfar 959
Gold BasinNWA1465ChelyabinskNWA 4482
Prebiotic chemistry in space conditions: the role of radiation on
formamide system
Thank you for the attention!