Seminario Biología Molecular, Tercer Semestre Medicina, Universidad Pontificia Bolivariana

1. N a t a l i a F l ó r e z S a l a z a r
N a t a l i a F l ó r e z S a l a z a r
T h i r d S e m e s t e r
T h i r d S e m e s t e r
B o l i v a r i a n P o n t i f i c i a l U n i v e r s i t y
B o l i v a r i a n P o n t i f i c i a l U n i v e r s i t y

2. D i a b e t e s
D i a b e t e s
M e l l i t u s
M e l l i t u s
Introduction
Introduction
A t r i a l
A t r i a l
F i b r i l l a t i o n
F i b r i l l a t i o n
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It is a group of
It is a group of
diseases that results
diseases that results
in too much sugar in
in too much sugar in
the blood (high
the blood (high
blood glucose).
blood glucose).
Irregular and occasionally
Irregular and occasionally
rapid heart rate that often
rapid heart rate that often
causes poor blood supply.
causes poor blood supply.

3. T h e M A P K 1 0 g e n e i s a c r i t i c a l
T h e M A P K 1 0 g e n e i s a c r i t i c a l
r e g u l a t o r o f D M a s s o c i a t e d A F a n d
r e g u l a t o r o f D M a s s o c i a t e d A F a n d
m a y b e a p o t e n t i a l t h e r a p e u t i c
m a y b e a p o t e n t i a l t h e r a p e u t i c
a p p r o a c h t o t r e a t D M r e l a t e d A F .
a p p r o a c h t o t r e a t D M r e l a t e d A F .
Gen MAPK10
Gen MAPK10

4. General objective
General objective
U s i n g t i m e - s e r i e s R N A s e q u e n c e
U s i n g t i m e - s e r i e s R N A s e q u e n c e
a n a l y s i s , M A P K 1 0 i s i d e n t i f i e d a s a
a n a l y s i s , M A P K 1 0 i s i d e n t i f i e d a s a
c r i t i c a l g e n e i n d i a b e t e s m e l l i t u s -
c r i t i c a l g e n e i n d i a b e t e s m e l l i t u s -
i n d u c e d a t r i a l f i b r i l l a t i o n i n m i c e .
i n d u c e d a t r i a l f i b r i l l a t i o n i n m i c e .

5. Materials and methods
Materials and methods
V a r i a n t o f t h e p o l y m e r a s e c h a i n r e a c t i o n .
V a r i a n t o f t h e p o l y m e r a s e c h a i n r e a c t i o n .
U s e d t o a m p l i f y a n d s i m u l t a n e o u s l y q u a n t i f y t h e
U s e d t o a m p l i f y a n d s i m u l t a n e o u s l y q u a n t i f y t h e
D N A a m p l i f i c a t i o n p r o d u c t .
D N A a m p l i f i c a t i o n p r o d u c t .
I t w a s u s e d t o e x t r a c t t o t a l R N A f r o m t h e l e f t
I t w a s u s e d t o e x t r a c t t o t a l R N A f r o m t h e l e f t
a t r i u m a n d r e v e r s e t r a n s c r i b e d i t .
a t r i u m a n d r e v e r s e t r a n s c r i b e d i t .
P r o t e i n s w e r e e x t r a c t e d f r o m c a r d i a c t i s s u e s a n d
P r o t e i n s w e r e e x t r a c t e d f r o m c a r d i a c t i s s u e s a n d
m e m b r a n e s w e r e i n c u b a t e d w i t h a n t i b o d i e s .
m e m b r a n e s w e r e i n c u b a t e d w i t h a n t i b o d i e s .
T h e p u r p o s e o f t h i s t e s t i s t o i d e n t i f y p r o t e i n s
T h e p u r p o s e o f t h i s t e s t i s t o i d e n t i f y p r o t e i n s
b a s e d o n t h e a n t i g e n - a n t i b o d y r e a c t i o n .
b a s e d o n t h e a n t i g e n - a n t i b o d y r e a c t i o n .
R e a l - t i m e P C R A n a l y s i s
R e a l - t i m e P C R A n a l y s i s W e s t e r n B l o t A n a l y s i s
W e s t e r n B l o t A n a l y s i s
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6. A s o l u t i o n w a s i n j e c t e d i n t o t h e t a i l v e i n o f
A s o l u t i o n w a s i n j e c t e d i n t o t h e t a i l v e i n o f
m i c e a t 1 0 w e e k s .
m i c e a t 1 0 w e e k s .
I t i s u s e d i n g e n e t h e r a p y p r o t o c o l s , s i n c e t h e y
I t i s u s e d i n g e n e t h e r a p y p r o t o c o l s , s i n c e t h e y
h a v e c h a r a c t e r i s t i c s t h a t m a k e t h e m t o o l s f o r
h a v e c h a r a c t e r i s t i c s t h a t m a k e t h e m t o o l s f o r
t h e t r e a t m e n t o f g e n e t i c d i s e a s e s .
t h e t r e a t m e n t o f g e n e t i c d i s e a s e s .
T h e a p o p t o s i s o f c a r d i o m y o c y t e s i n h e a r t
T h e a p o p t o s i s o f c a r d i o m y o c y t e s i n h e a r t
s e c t i o n s w a s e v a l u a t e d u s i n g t h i s t e s t .
s e c t i o n s w a s e v a l u a t e d u s i n g t h i s t e s t .
T h e a s s a y i s b a s e d o n t h e p r e s e n c e o f b r e a k s i n
T h e a s s a y i s b a s e d o n t h e p r e s e n c e o f b r e a k s i n
t h e D N A w h i c h c a n b e i d e n t i f i e d b y t h e T d T
t h e D N A w h i c h c a n b e i d e n t i f i e d b y t h e T d T
t e r m i n a l .
t e r m i n a l .
I n j e c t i o n o f R e c o m b i n a n t
I n j e c t i o n o f R e c o m b i n a n t
A d e n o - A s s o c i a t e d V i r u s
A d e n o - A s s o c i a t e d V i r u s
T u n n e l A s s a y
T u n n e l A s s a y
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7. Results
Results
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MAPK10 (involved gene), TNF α (cytokine), and tubulin (test
MAPK10 (involved gene), TNF α (cytokine), and tubulin (test
control) are associated with WT and db/db mice followed by
control) are associated with WT and db/db mice followed by
follow-up at week 12, 14, and 16.
follow-up at week 12, 14, and 16.
This image shows an experimental model where a protein is
This image shows an experimental model where a protein is
measured and as time passes, its expression increases and,
measured and as time passes, its expression increases and,
likewise, the damage of the material is evaluated.
likewise, the damage of the material is evaluated.

8. ______________________________________________________
In this graph they show us an immunoassay where the main
In this graph they show us an immunoassay where the main
focus is on showing how in the case of Kv4.3 it is intensified in
focus is on showing how in the case of Kv4.3 it is intensified in
non-mutated mice, Kv4.2 is decreased in mutated mice.
non-mutated mice, Kv4.2 is decreased in mutated mice.
ITNFα is intensified in the db/db mutated mice, as is the
ITNFα is intensified in the db/db mutated mice, as is the
MAPK10 gene and tubulin. It can also be seen how p-stat3,
MAPK10 gene and tubulin. It can also be seen how p-stat3,
which is a mediator of the signaling process, increases
which is a mediator of the signaling process, increases
equally in db/db mice.
equally in db/db mice.

9. ______________________________________________________
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Discussion
Discussion
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Author
Author What did he say
What did he say Agree or not
Agree or not
Wang, et al.
Wang, et al.
Liu, et al.
Liu, et al.
Heijman, et al.
Heijman, et al.
Increased AF susceptibility in
Increased AF susceptibility in
db/db mice occurred in
db/db mice occurred in
association with atrial electrical
association with atrial electrical
and estructural remodeling.
and estructural remodeling.
A previous study showed that
A previous study showed that
hyperglycemia contributes to atrial
hyperglycemia contributes to atrial
interstitial fibrosis in an animal
interstitial fibrosis in an animal
model.
model.
Increased atrial fibrosis slows atrial
Increased atrial fibrosis slows atrial
conduction and leads to
conduction and leads to
asynchronous or disorganized
asynchronous or disorganized
conduction.
conduction.

10. Conclusions
Conclusions
M e d i c a l s t u d i e s ( s u c h a s t h i s
M e d i c a l s t u d i e s ( s u c h a s t h i s
o n e ) w o u l d n o t b e p o s s i b l e
o n e ) w o u l d n o t b e p o s s i b l e
w i t h o u t t h e g r a p h i c s a n d
w i t h o u t t h e g r a p h i c s a n d
a p p l i c a t i o n s t h a t a r e s h o w n
a p p l i c a t i o n s t h a t a r e s h o w n
c o r r e s p o n d i n g t o s t u d i e s o f
c o r r e s p o n d i n g t o s t u d i e s o f
i m m u n o a s s a y s .
i m m u n o a s s a y s .
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M o l e c u l a r b i o l o g y p r o v i d e s v e r y
M o l e c u l a r b i o l o g y p r o v i d e s v e r y
i m p o r t a n t s t u d i e s f o r t h e
i m p o r t a n t s t u d i e s f o r t h e
a d v a n c e m e n t o f m e d i c i n e a n d i t s
a d v a n c e m e n t o f m e d i c i n e a n d i t s
r e s e a r c h .
r e s e a r c h .