prezentácia programu powerpoint - imbm · extracelulárnadna mandel p, metais p. les acides...
TRANSCRIPT
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Plazma pod mikroskopom
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Extracelulárna DNA
Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243.
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Extracelulárna DNA
Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243.
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Extracelulárna DNA
NIPD
CVS
Cancer
![Page 9: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/9.jpg)
Extracelulárna DNA
Lo YM et al.: Presence of fetal DNA in maternal plasma and serum. Lancet 1997; 350(9076):485-7.
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Fetálna DNA v maternálnej plazme
• Aplikácie• Diagnostika fetálneho pohlavia
• SRY, DYS, XX?
• Diagnostika fetálneho RhD statusu
• Aneuploídie
• Monogénne podmienené poruchy
• ...
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Fetálna DNA & preeklampsia
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Príčina→Dôsledok?
?Dôsledok←Príčina
Fetálna DNA
Preeklampsia
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Fetálna DNA & preeklampsia
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Fetálna DNA & preeklampsia
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P l a c e n t a l w e i g h t
C T R L a d u l t D N A
m o u s e
f e t a l D N A
m o u s e
C p G f e t a l D N A
h u m a n
L P S
0 . 0 0
0 . 0 5
0 . 1 0
0 . 1 5
g
* * *
F e t a l w e i g h t
C T R L a d u l t D N A
m o u s e
f e t a l D N A
m o u s e
C p G f e t a l D N A
h u m a n
L P S
0 . 0
0 . 5
1 . 0
1 . 5
2 . 0
g
*
L i t t e r s i z e
C T R L a d u l t D N A
m o u s e
f e t a l D N A
m o u s e
C p G f e t a l D N A
h u m a n
L P S
0
5
1 0
1 5
nu
mb
er
of f
etu
se
s
* *
Reprodukovateľnosť
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P l a c e n t a l w e i g h t
C T R L a d u l t D N A
m o u s e
f e t a l D N A
m o u s e
C p G f e t a l D N A
h u m a n
L P S
0 . 0 0
0 . 0 5
0 . 1 0
0 . 1 5
g
* * *
F e t a l w e i g h t
C T R L a d u l t D N A
m o u s e
f e t a l D N A
m o u s e
C p G f e t a l D N A
h u m a n
L P S
0 . 0
0 . 5
1 . 0
1 . 5
2 . 0
g
*
L i t t e r s i z e
C T R L a d u l t D N A
m o u s e
f e t a l D N A
m o u s e
C p G f e t a l D N A
h u m a n
L P S
0
5
1 0
1 5
nu
mb
er
of f
etu
se
s
* *
Reprodukovateľnosť
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Je fetálna DNA vyššia pri preeklampsii?
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Čo môže byť príčinou vyššej ecDNA?
• Nefunkčné odstraňovanie• Poruchy pečene
• Nižšia DNázová aktivita
• Rezistencia voči fragmentácii
• ...
• Vyššia produkcia• Poškodenie tkaniva
• NETóza
• ...
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Pôvod CNA
• DNA/RNA
• Voľné/viazané na proteíny/viazané na povrch buniek
• Nekróza/apoptóza/aktívna sekrécia?
• Hematopoetické bunky/ostatné tkanivá/nádory-plody
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Fragmentácia CNA
• Vysoká fragmentácia ecDNA
• Nukleozomálny rebrík
• Fragmenty – n x 200 bp
• Deoxyribonukleázy v krvi
• Sekrécia do moča
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Akútny infarkt myokardu & ecDNA
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Akútny infarkt myokardu & ecDNA
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NCMP & ecDNA
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Maratón & ecDNA
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ecDNA v onkológii
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• Dlhšie fragmenty – nekróza?
• Kratšie fragmenty – ctDNA
• mtDNA vyššia
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Genometastáza
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Experiment
• Thioacetamide (TAA)• 2x300 mg/kg
TAA 24h TAA 6h
![Page 45: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/45.jpg)
PečeňA S T
k
at/
l
C T R L T AA T AA+ D N a s e
0
5 0
1 0 0
1 5 0
**
*
G M T
k
at/
l
C T R L T AA T AA+ D N a s e
0 .0 0
0 .0 1
0 .0 2
0 .0 3
0 .0 4
0 .0 5 ***
*
B iliru b in
m
ol/
l
C T R L T AA T AA+ D N a s e
0
5
1 0
1 5
2 0
*
A L T
k
at/
l
C T R L T AA T AA+ D N a s e
0
5
1 0
1 5
2 0
**
![Page 46: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/46.jpg)
Obličky
C re a t in in e
m
ol/
l
C T R L T AA T AA+ D N a s e
0
2 0
4 0
6 0
8 0 **
+
B U N
mm
ol/
l
C T R L T AA T AA+ D N a s e
0
5
1 0
1 5
**
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T o t a l e x t r a c e l l u l a r D N A
ng
/ml
pla
sm
a
C T R L T A A T A A + D N a s e
0
1 0 0 0 0
2 0 0 0 0
3 0 0 0 0
4 0 0 0 0
* *
EcDNA
N u c l e a r D N A
GE
/ml
pla
sm
a
C T R L T A A T A A + D N a s e
0
5 0 0 0 0
1 0 0 0 0 0
1 5 0 0 0 0
2 0 0 0 0 0 **
M i t o c h o n d r i a l D N A
ng
/ml
pla
sm
a
C T R L T A A T A A + D N a s e
0 . 0 0
0 . 0 2
0 . 0 4
0 . 0 6
*
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M y e l o p e r o x i d a s e a c t i v i t y
mU
/ml
C T R L T A A T A A + D N a s e
0 . 0
0 . 1
0 . 2
0 . 3
0 . 4
*
Aktivácia neutrofilov
![Page 49: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/49.jpg)
Príčina→Dôsledok?
?Dôsledok←Príčina
EcDNA
Poškodenie pečene
![Page 50: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/50.jpg)
Experiment
• Thioacetamide (TAA)• 2x300 mg/kg
• Deoxyribonukleáza (DNase)• 2x10 mg/kg
TAA 24h TAA 6h
DNase DNase
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Plazmatická DNáza
C T R L T A A T A A + D N a s e
0 . 0 0
0 . 0 1
0 . 0 2
0 . 0 3
0 . 0 4
0 . 0 5
P l a s m a D N a s e a c t i v i t y
Ku
nit
z u
nit
s
* *
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T o t a l e x t r a c e l l u l a r D N A
ng
/ml
pla
sm
a
C T R L T A A T A A + D N a s e
0
1 0 0 0 0
2 0 0 0 0
3 0 0 0 0
4 0 0 0 0
* *
EcDNA
N u c l e a r D N A
GE
/ml
pla
sm
a
C T R L T A A T A A + D N a s e
0
5 0 0 0 0
1 0 0 0 0 0
1 5 0 0 0 0
2 0 0 0 0 0 **
M i t o c h o n d r i a l D N A
ng
/ml
pla
sm
a
C T R L T A A T A A + D N a s e
0 . 0 0
0 . 0 2
0 . 0 4
0 . 0 6
*
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M y e l o p e r o x i d a s e a c t i v i t y
mU
/ml
C T R L T A A T A A + D N a s e
0 . 0
0 . 1
0 . 2
0 . 3
0 . 4
*
Aktivácia neutrofilov
![Page 54: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/54.jpg)
PečeňA S T
k
at/
l
C T R L T AA T AA+ D N a s e
0
5 0
1 0 0
1 5 0
**
*
G M T
k
at/
l
C T R L T AA T AA+ D N a s e
0 .0 0
0 .0 1
0 .0 2
0 .0 3
0 .0 4
0 .0 5 ***
*
B iliru b in
m
ol/
l
C T R L T AA T AA+ D N a s e
0
5
1 0
1 5
2 0
*
A L T
k
at/
l
C T R L T AA T AA+ D N a s e
0
5
1 0
1 5
2 0
**
![Page 55: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/55.jpg)
Obličky
C re a t in in e
m
ol/
l
C T R L T AA T AA+ D N a s e
0
2 0
4 0
6 0
8 0 **
+
B U N
mm
ol/
l
C T R L T AA T AA+ D N a s e
0
5
1 0
1 5
**
![Page 56: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/56.jpg)
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ecDNA & Trauma
![Page 60: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/60.jpg)
ecDNA & Trauma
![Page 61: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/61.jpg)
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Nukleárna ecDNA
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Nukleárna ecDNA
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Nukleárna ecDNA
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C T R L IC P
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
T o ta l e x tra c e llu la r D N A
ng
/ml
pla
sm
a
C T R L IC P
0
5 0 0
1 0 0 0
1 5 0 0
N u c le a r D N A
GE
/ml
pla
sm
a
*
C T R L IC P
0
5 0 0 0 0
1 0 0 0 0 0
1 5 0 0 0 0
M ito c h o n d r ia l D N A
GE
/ml
pla
sm
a
Intrahepatálna cholestáza tehotných
![Page 73: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/73.jpg)
Nuleárna ecDNA
C T R L IC P
0
2 0 0 0
4 0 0 0
6 0 0 0
8 0 0 0
N u c le a r D N A - s h o r t fra c tio n
GE
/ml
pla
sm
a
**
C T R L IC P
0
5 0 0
1 0 0 0
1 5 0 0
2 0 0 0
2 5 0 0
N u c le a r D N A - lo n g f ra c tio n
GE
/ml
pla
sm
a
C T R L IC P
0
2
4
6
8
F ra g m e n ta t io n in d e x
AU
*
![Page 74: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/74.jpg)
Preeklampsia
C T R L P E
0
2 0 0
4 0 0
6 0 0
T o ta l e x tra c e llu la r D N A
ng
/ml
pla
sm
a
**
C T R L P E
0
1 0 0
2 0 0
3 0 0
4 0 0
N u c le a r D N A
GE
/ml
pla
sm
a
***
C T R L P E
0
5 0 0 0
1 0 0 0 0
1 5 0 0 0
2 0 0 0 0
2 5 0 0 0
M ito c h o n d r ia l D N A
GE
/ml
pla
sm
a
![Page 75: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/75.jpg)
Preeklampsia
C T R L P E
0
5 0 0
1 0 0 0
1 5 0 0
N u c le a r D N A - s h o r t fra c tio n
GE
/ml
pla
sm
a
**
C T R L P E
0
2 0 0
4 0 0
6 0 0
N u c le a r D N A - lo n g f ra c tio n
GE
/ml
pla
sm
a
C T R L P E
0
2 0
4 0
6 0
8 0
1 0 0
F e ta l D N A - s h o rt fra c tio n
GE
/ml
pla
sm
a
C T R L P E
0
1 0
2 0
3 0
F e ta l D N A - lo n g f ra c tio n
GE
/ml
pla
sm
a
C T R L P E
0
1
2
3
4
5
N u c le a r D N A fra g m e n ta t io n
AU
*
![Page 76: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/76.jpg)
CTRL Mitochondria Sepsis
![Page 77: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/77.jpg)
Circulus vitiosus
Poškodenie tkaniva,
Infekcia, ...
NETóza
TLR9
Vrodená imunita Preeklampsia,
Hepatopatie, Sepsa, Lupus, RA
....
![Page 78: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/78.jpg)
Circulus vitiosus
Poškodenie tkaniva,
Infekcia, ...
NETóza
TLR9
Vrodená imunita Preeklampsia,
Hepatopatie, Sepsa, Lupus, RA
....
AspirínImunosupresíva
DNase
Mg2+
Metformín
![Page 80: Prezentácia programu PowerPoint - IMBM · ExtracelulárnaDNA Mandel P, Metais P. Les acides nucleiques du plasma sanguin chez l'homme. C R Acad Sci Paris 1948; 142:241-243](https://reader033.vdocuments.net/reader033/viewer/2022041809/5e5653255009135ff4302a5b/html5/thumbnails/80.jpg)
Financial efficiency