Facultad de medicina Universidad Pontificia Bolivariana

1. Miguel Saavedra Valencia.
3° semestre / Facultad de Medicina / Universidad Pontificia Bolivariana

2. CARDIOVASCULAR DISEASES
Recommendations
The most common…
1. Coronary artery disease
(narrowing of the arteries).
2. Heart attack !
3. Abnormal heart rhythms, or
arrhythmias.
4. Heart failure.
5. Heart valve disease.
*the symptoms depend on the type
of disease*
What are the cardiovascular diseases?
Range of conditions that affect your
heart, involve narrowed or blocked blood
vessels and can lead to a heart attack,
chest pain (angina) or stroke.
1
CVD 2
3
17.9 million people died from
CVDs in 2016, representing
31% of all global deaths.
20.728 People in Colombia
• Eat a balanced diet
• Build healthy lifestyle habits
• Visit your doctor frequently

3. cfDNA DNasa I MICROCHIPS
Is a DNA or RNA free in
blood stream
Originates from the
apoptosis or necrosis of all
cell types.
Employing as a liquid biopsy,
providing access to the
entire tumor genome.
Elevated cfDNA
concentration in myocardial
infarction (MI) patients
Is the primary factor
responsable for regulating
the elimination and
degradation of cfDNA from
the blood stream
(pancreas and parotid)
Endonuclease that attacks
double-stranded DNA. This
enzyme requires divalent
cations (Ca2+ and Mg2+)
and is active at neutral pH
suggested to be involved in
internucleosomal DNA
degradation during
apoptosis
is a collection of
microscopic DNA
spots attached to a
solid surface.
are used to determine
the differential
expression of genes in
various circumstances,
for example, The
presence or absence of
DNA in a disease

4. OBJECTIVE
Perform fragment analysis on plasma cfDNA
from cardiac disease patients using microchip
electrophoresis and to investigate the
relationships between DNase I activity and
cfDNA concentration.

5. POBLACIÓN
MÉTODOS

6. EXTRACIÓN DEL DNA
1. Lisis celular 2. RNAasa (opcional)
3. Precipitación de
proteínas con ácido
acético
4. Precipitación del
ADN con isopropanol 5. Almacenaje en TE
(Tris y EDTA)
6. Medición de
absorbancia en
espectrofotómetro
(260nm-280nm)
MÉTODOS

7. MICROCHIPS DE
ELECTROFORESIS
Microchip de electroforesis
Es rápida y sensible, da como resultado una alta
resolución y permite una fácil separación del ADN.
Promete un consumo mínimo de muestras y
reactivos, un tiempo de análisis corto, eficiencia,
integración y automatización
¿Para qué se hace?
Separa proteínas y moléculas como ADN y
ARN.
Fundamento
migración proporcional de las moléculas a
través de un gel u otro tipo de matriz porosa,
según su peso molecular o tamaño;
movimiento generado por el campo eléctrico
MÉTODOS

8. ACTIVIDAD
DE LA DNasa
¿Para qué se hace?
Conocer la cantidad
cfDNA.
Se encarga de
eliminar el
cfDNA de la
sangre.
Fundamento
(SRED) single radial
enzyme diffusion
MÉTODOS

9. RESULTADOS
PACIENTES GRUPO CONTROL
En muestras de control
sanas, se observó una
región de banda única de
150-200 pb en algunos
carriles, pero no se
observaron fragmentos en
algún sujeto sano

10. RESULTADOS
Se observaron tres
fragmentos (150–200 pb,
300–400 pb, y 500–600 pb)
en todos los pacientes con
IM, angina cardíaca y todas
las demás muestras de
pacientes cardíacos.
PACIENTES CON
INFARTO DE MIOCARDIO

11. RESULTADOS
Se observaron tres
fragmentos (150–200 pb,
300–400 pb, y 500–600 pb)
PACIENTES CON ANGINA
CARDIACA

12. RESULTADOS
Los resultados indican que una
proporción de 150 a 200 pb /
500 a 600 pb en el IM fue
significativamente mayor que
la de otros pacientes con
enfermedad cardíaca (dolor de
pecho, angina cardíaca,
fibrilación auricular e
insuficiencia cardíaca)

13. DISCUSSION
AUTHOR CONCEPT YES OR NOT
CP Chang, et al.
a>10-fold elevation of
cfDNA in MI patients
when compared to normal
control individuals
Ershova et al.
Described a negative
correlation between cfDNA
concentration and DNase I
activity in blood of healthy
non-pregnant women
E. Heitzer et al.
Apoptosis produces
fragments that are the
same size as a typical
ladder pattern (150–200 bp,
300–400 bp, and 500–600
bp)

14. CONCLUSIONS
There is a positive correlation between
DNase I activity and cfDNA concentration,
because when one rise the other one does
too.
cfDNA levels are acutely elevated in MI, for
that reason the cfDNA and DNase I may
serve as a potential diagnostic and
prognostic biomarker for MI and may have
advantage for early diagnosis
Currently, the applications of molecular
biology offer to health professionals the
tools that allow them to investigate
pathologies, achieving to diagnose
diseases and its adequate treatment.
All the results of this research are thanks to
the molecular biology.