Radiation Chemistry; Effects of Radiation On DNA and ... · DNA is a Primary Target • Microbeam experiments show cell nucleus to be more sensitive than cytoplasm. • Halogenated

1

Radiation Chemistry

Effects of Radiation On DNA and

Chromosomes

Kathryn D Held PhD

Massachusetts General Hospital

Harvard Medical School

2


Chromosomes

bull Introduction to Radiation Chemistry

bull Water Radiolysis

bull DNA Damage

bull Chromosome Aberrations

3

Sequences in the Development of Radiobiological Effects

Time Event

10-18

s Absorption of Ionizing Radiation

10-16

s Physical Events

Ionization

Excitation

10-12

s Physicochemical Events

Free radical formation

Breakage of chemical bonds

10-12

ndash 10-6

s Chemical Events

Reactions of radicals

Minutes to hours BiochemicalCellular Processes

Repair

Division delay

Chromosome damage

Loss of reproductive capacity

Days to months Tissue Damage

CNS GI Bone marrow syndromes

Late tissue damage

Birth defects from in utero exposure

Years Late Somatic Effects

Cataracts

Carcinogenesis

Generations Genetic Effects

4

Ionizing Radiation

bull All biological effects produced by ionizing

radiation result from the chemical events that

occur shortly after the initial deposition of

radiation energy

bull Absorption of IR by matter produces ions and

excited molecules

A A+ + e-

A A

bull The number of species produced is proportional

to dose

5

Ionizing Radiation

bull Free radicals - atoms or molecules that

have one or more unpaired electron

ndash designated by ldquobullrdquo

ndash may be formed by division of a covalent

bond

RS Rbull + Sbull

ndash may be charged or neutral

ndash are generally very reactive

6

Direct and Indirect Actions of

Ionizing Radiation (from Hall 1994)

Low LET

70

30

7


Chromosomes



bull DNA Damage


8

Water Radiolysis Summary

H2O bullOH bullH e-aq H2 H2O2

bullOH is the most important biologically

9

Mechanisms of Water Radiolysis

Ionization and Excitation

H2O H2O+ + e-

H2O H2O

Ion-molecule interaction and dissociation

H2O+ + H2O H3O

+ + bullOH

e- + H2O bullH + OH-

H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq

Spur reactionsbullH + bullH H2

bullOH + bullOH H2O2

bullOH + bullH H2O

Diffusion

10

G Values (Yields) of Primary Radiolysis

Species in Neutral Water

-H2O bullbullbullbullOH

bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010

(G-value = moles of material formed or changed by an energy absorption of 1 J)

Note

bull For low LET radiation the most common radiolysis species are OH and

e-aq

bull With higher LET radiation yields of radical species decrease and

molecular species increase

11

Reactions of bullOH

bull Oxidation of inorganic compoundsbullOH + Mn OH- + Mn+1

bull Addition to free radicals and unsaturated

organicsbullOH + CH2=CH2

bullCH2-CH2OH

bull Abstraction from saturated organicsbullOH + CH3COCH3

bullCH2COCH3 + H2O

12

e-aq and bullH

bull Reducing species

bull Frequently undergo diffusion-controlled

reactions

bull Reactions do not seem to be biologically

damaging

13

Reactions of Primary Radicals

with Oxygen

bull Formation of perhydroxyl and

superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488

bull Reactions with organic radicals

O2 + Rbull RO2bull

Note These reactions are thought to be responsible for

the Oxygen Effect

14

Radical Scavenging

bull Use of a compound that selectively reacts

with certain free radicals

bull Simplifies more complex radiation

chemistry

15

Radical Scavengers

Additive Reaction Active

Species

Remaining

N2O N 2O + e-

aq + H 2O

rarrrarrrarrrarr bullbullbullbullOH + OH

- + N 2

bullbullbullbullOH (H

bullbullbullbull)

bullbullbullbullOH

scavengers

RH + bullbullbullbullOH rarrrarrrarrrarr R

bullbullbullbull +

H 2O e

-

aq (Hbullbullbullbull)

oxygen O 2 + e-

aq rarrrarrrarrrarr O 2

-bullbullbullbull

O 2 + bullbullbullbullH rarrrarrrarrrarr HO 2

bullbullbullbull

bullbullbullbullOH O 2

bullbullbullbull-

HO 2bullbullbullbull

acid e-

aq + H+ rarrrarrrarrrarr Hbullbullbullbull

Hbullbullbullbull

bullbullbullbullOH

16

bullOH Scavengers Decrease Radiation-

induced DNA Damage (from Roots and Okada 1972)

17


Chromosomes



bull DNA Damage


18

DNA is a Primary Target

bull Microbeam experiments show cell nucleus to be more sensitive than cytoplasm

bull Halogenated base analogues sensitize cells and DNA

bull Radioisotopes in DNA are more lethal than when in RNA or protein

bull DNA repair deficient cells are radiation sensitive drugs that inhibit DNA repair usually are radiosensitizers

bull Oxygen and LET modify survival cytogeneticdamage and biological activity of DNA in similar manner

19

Reactions of bullOH with DNA Bases and Sugar

20

Reactions of bullOH with DNA Bases and

Sugar

bull Subsequent to radical production in

DNA a multitude of products can be

formed

bull Eg from thymine alone more than 30

radiolysis products have been identified

with quite different yields

21

Types of DNA Lesions from IR

From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change

bull Loss (abasic sites)

Crosslinks

bull DNA-DNA

bull DNA-protein

22

Measurement of DNA Damages

bull Base damages

ndash Enzyme sensitivity

ndash HPLC GC-MS GC-EC

ndash Immunological probes

bull Strand breaks

ndash Gel electrophoresis (alkaline for SSB

neutral for DSB)

ndash Comet assay (alkaline for SSB neutral for

DSB)

ndash Foci of DNA repair-related proteins(eg γγγγ-H2AX)

23

Pulsed Field Gel Electrophoresis

24

Comet Assay (Single Cell Gel Electrophoresis)

bull Embed cells in gel and lyse

bull Electrophorese

bull Quantify amount of DNA in ldquotailrdquo (damaged) versus ldquoheadrdquo

25

Comparison of Assays for Breaks

(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose

bull Killing usually

shouldered

26

Foci of DNA Repair-Related Proteins (eg

γγγγ-H2AX) as Measure of DNA DSBs

(from Bonner 2003)

γγγγ-H2AX ndash phosphorylated histone H2A variant X

Foci ndash fluorescent ldquoblobsrdquo representing aggregates of protein

recognized by antibodies

27

γγγγ-H2AX Foci as a Measure of DSBs

(from Rothkamm and Loumlbrich 2003)

(o = focicell ∆ = PFGE)

slope = 35 DSBcellGyNote

Number of foci increases linearly with dose with same slope

as DSBs measured by PFGE

Even after very low doses some foci remain at 24 h

28

Foci as Measure of DSBs

bull Caution γγγγ-H2AX foci have been seen

after treatments that do not directly

cause DSBs eg hypoxia hydrogen

peroxide

bull Other DNA repair-related proteins also

form foci and are being used as

surrogates for DSBs eg 53BP1

RAD51

29

Number of Radiation-Induced Lesions

Type of Lesion Number per Gray

Double strand breaks 40

Single strand breaks 1000

Base damages 1000-2000

Sugar damages 800-1000

DNA-DNA crosslinks 30

DNA-protein crosslinks 150

Alkali-labile sites 200-300

Number of Clustered Lesions not yet quantified

30

Energy Deposition Events (spurs

blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV

Branch TracksDelta rays

Blobs

Short Tracks

Spurs

(from Mozunder and Magee 1966)

bullbull bullbullbullbullbullbull

bull bullbullbullbullbullbullbull

bullbullbull

bull

bullbullbull

bullbull bullbull

31

Energy Deposition Events for Low

LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5

Short track 500-5000 eV

DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm

Electrons with sufficient energy to form short tracks will also produce spurs and blobs (from

Ward 1988)

32

Clustered Lesions (Multiply

Damaged Sites) (from Steel 1993)

33

Biological Consequences of

Clustered Lesions (MDS)

bull Harder to repair accurately than single

lesions

bull Unrepairedndash Block DNA replication

ndash Loss of genetic integrity

bull Mispaired

ndash May lead to DSBs

ndash Deletions could be produced

bull Repair could be completed accurately

34

Measured Clustered Lesions

Relative Cluster Frequencies

in Human Cells

DSB 1

Oxidized purines 1

Oxidized pyrimidines 09

Abasic sites 075

(from Sutherland et al 2002)

35

Chromatin Structure is Important in Radiation Damage to DNA

bull Presence of histones

chromatin

condensation

bull Regionally multiply

damaged sites

bull Actively transcribing

vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36

Importance of HistonesChromatin

Condensation

Recent studies suggest histone deacetylase (HDAC)

inhibitors may be radiation sensitizers

(from Campausen et al 2004a) (from Campausen et al 2004b)

37

Which DNA lesion is most important

biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38

Most Important Lesion

bull To date most data suggest DSBs

bull Most assays for DSBs will include

Clustered Lesions

bull Clustered Lesions may be most

important for cell killing

39

Biological Consequences

DNA damage

Accurate repair Misrepair No repair

Mutations

Chromosome aberrations

Genomic instability

Neoplastic transformation

Cell deathinactivation

Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41

Chromosome Aberrations

bull Reflect

ndash initial DNA damage

ndash its repair (or nonmisrepair)

bull Two general types

ndash Chromosome aberrations

bull G1 irradiation

bull Both sister chromatids involved

ndash Chromatid aberrations

bull S or G2 irradiation

bull Usually only one chromatid involved

42

Chromosome versus Chromatid Aberrations (from McMillan and Steel 1993)

43

Examples of Chromosome Aberrations

dicentrics

tricentric

fragment

ring

44

Examples of Chromatid Aberrations

quadra-radials

complex

exchange

45


bull Principal aberrations

ndash Dicentrics

ndash Rings

ndash Acentric fragments

ndash Translocations

ndash Anaphase bridges

bull Exchange-type aberrations can be symmetric or asymmetric

bull Aberrations can be stable or unstable

bull Dicentrics (eg in lymphocytes) are a good biomarker of radiation exposure

46

Micronuclei formation is sometimes used as

a surrogate for chromosome aberrations

Micronuclei can

result from

chromosome

deletions or

fragments

47

Techniques Used in Chromosome

Analysis

bull Premature Chromosome Condensation (PCC)

bull Fluorescence in Situ Hybridization (FISH chromosome ldquopaintingrdquo)

Use of FISH-based techniques has made it clear that

bull radiation-induced chromosome aberrations are more complex than previously realized

bull complexity of aberrations increases with LET

48

Example of mFISH

Metaphase chromosomes

from lymphocytes of

plutonium-exposed

individual showing

complex rearrangements (from Anderson et al 2005)

49

Dose Response Curve for Chromosome

Aberrations is Linear-Quadratic

(From Hall 2000)

50

Good Correlation Between Chromosome Aberrations and Loss of Clonogenicity

(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52

Take Home Messages - 1

bull Indirect action produces most damage from low LET radiation bullOH is the most critical water radiolysis species

bull A plethora of DNA damages are produced by IR

ndash Numerous techniques can be used to measure the damage

ndash Using foci of DNA repair-related proteins to measure DSBs is currently of great interest

bull IR produces clustered lesions (multiply damaged sites) that are probably most important biologically

53


bull Chromatin structure is important for radiation

damage to DNA and its repair

bull The biological consequences of misrepair or no

repair include mutations aberrations genomic

instability cell deathinactivation

bull It is assumed that most cell death results from

DNA damage relationships between loss of

clonogenicity apoptosis or long-term arrest are

not straightforward

2


Chromosomes



bull DNA Damage


3


Time Event

10-18


10-16

s Physical Events

Ionization

Excitation

10-12




10-12

ndash 10-6

s Chemical Events



Repair

Division delay

Chromosome damage




Late tissue damage



Cataracts

Carcinogenesis


4

Ionizing Radiation




radiation energy


excited molecules

A A+ + e-

A A


to dose

5

Ionizing Radiation





bond

RS Rbull + Sbull



6



Low LET

70

30

7


Chromosomes



bull DNA Damage


8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

3


Time Event

10-18


10-16

s Physical Events

Ionization

Excitation

10-12




10-12

ndash 10-6

s Chemical Events



Repair

Division delay

Chromosome damage




Late tissue damage



Cataracts

Carcinogenesis


4

Ionizing Radiation




radiation energy


excited molecules

A A+ + e-

A A


to dose

5

Ionizing Radiation





bond

RS Rbull + Sbull



6



Low LET

70

30

7


Chromosomes



bull DNA Damage


8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

4

Ionizing Radiation




radiation energy


excited molecules

A A+ + e-

A A


to dose

5

Ionizing Radiation





bond

RS Rbull + Sbull



6



Low LET

70

30

7


Chromosomes



bull DNA Damage


8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

5

Ionizing Radiation





bond

RS Rbull + Sbull



6



Low LET

70

30

7


Chromosomes



bull DNA Damage


8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

6



Low LET

70

30

7


Chromosomes



bull DNA Damage


8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

7


Chromosomes



bull DNA Damage


8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

8




9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

9



H2O H2O+ + e-

H2O H2O


H2O+ + H2O H3O

+ + bullOH


H2O bullH + bullOH

Electron hydration

e- + (H2O)n e-aq



bullOH + bullH H2O

Diffusion

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

10




bullbullbullbullH e

-aq H2 H2O2

γγγγ-rays and

electrons of

01 ndash 20 MeV

043 028 006 028 005 007

32 MeV αααα-

particles031 009 004 008 010 010


Note


e-aq



11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

11

Reactions of bullOH




bullCH2-CH2OH


bullCH2COCH3 + H2O

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

12

e-aq and bullH



reactions


damaging

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

13


with Oxygen


superoxide radicals

O2 + bullH HO2bull

O2 + e-aq O2

-bull

O2-bull + H+ HO2

bull pK = 488


O2 + Rbull RO2bull


the Oxygen Effect

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

14

Radical Scavenging




chemistry

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

15

Radical Scavengers


Species

Remaining

N2O N 2O + e-

aq + H 2O


- + N 2


bullbullbullbull)

bullbullbullbullOH

scavengers


bullbullbullbull +

H 2O e

-


oxygen O 2 + e-


-bullbullbullbull


bullbullbullbull


bullbullbullbull-


acid e-


Hbullbullbullbull

bullbullbullbullOH

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

16



17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

17


Chromosomes



bull DNA Damage


18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

18







19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

19


20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

20


Sugar



formed




21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

21


From McMillan

and Steel 1993

Breaks

bull SSB

bull DSB

Base damages

bull Change


Crosslinks

bull DNA-DNA

bull DNA-protein

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

22


bull Base damages




bull Strand breaks


neutral for DSB)


DSB)


23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

23


24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

24



bull Electrophorese


25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

25


(from Olive 1992)

Note

bull More SSBs

than DSBs

bull Breaks usually

linear with

dose


shouldered

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

26



(from Bonner 2003)




27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

27








28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

28





peroxide




RAD51

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

29











30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

30


blobs short tracks)

100 to 500 eV

lt 100 eV lt5000 eV

Primary

gt5000 eV


Blobs

Short Tracks

Spurs




bullbullbull

bull

bullbullbull

bullbull bullbull

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

31


LET Radiation

ENTITY

ENERGY

DEPOSITED SIZE

NUMBER OF

WATER

MOLECULES

PER EVENT

ENERGY

()

EVENTS

()

Spur lt100 eV 4 nm (diam) 1100 ~80 95

Blob lt500 eV 7 nm (diam) 6000 ~20 5


DNA 2 nm (diam)

Nucleosome

disc

Thickness 57 nm

Radius of 55 nm


Ward 1988)

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

32



33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

33




lesions



bull Mispaired




34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

34



in Human Cells

DSB 1

Oxidized purines 1


Abasic sites 075


35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

35



chromatin

condensation


damaged sites


vs non-transcribing

DNA

bull Nuclear matrix

attachment sites

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

36


Condensation




37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

37


biologically

Good correlation

between DNA DSBs

and cell killing

(from Radford 1985)

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

38




Clustered Lesions



39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

39


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

40


Chromosomes



bull DNA Damage


41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

41


bull Reflect





bull G1 irradiation





42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

42


43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

43


dicentrics

tricentric

fragment

ring

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

44


quadra-radials

complex

exchange

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

45



ndash Dicentrics

ndash Rings







46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

46



Micronuclei can

result from

chromosome

deletions or

fragments

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

47


Analysis






48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

48

Example of mFISH


from lymphocytes of

plutonium-exposed

individual showing


49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

49



(From Hall 2000)

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

50


(from Hall

2000)

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

51


DNA damage


Mutations


Genomic instability



Mitotic

Apoptotic

Long-term arrest

Survival

no mutations

52







53










not straightforward

52







53










not straightforward

53










not straightforward

Documents

Radiation Chemistry; Effects of Radiation On DNA and ... · DNA is a Primary Target • Microbeam experiments show cell nucleus to be more sensitive than cytoplasm. • Halogenated