1) Cardiovascular diseases (CVDs) are conditions that affect the heart and blood vessels. The most common CVDs are coronary artery disease, heart attack, arrhythmias, heart failure, and heart valve disease. 2) CVDs killed 17.9 million people worldwide in 2016, accounting for 31% of all global deaths. In Colombia, 20,728 people died from CVDs. 3) The study analyzed cell-free DNA (cfDNA) fragments in plasma samples from CVD patients using microchip electrophoresis. Three cfDNA fragment sizes (150–200 bp, 300–400 bp, and 500–600 bp) were observed in patients with myocardial infarction, angina, and other heart

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
MÉTOD
OS

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)
MÉTOD
OS

7. MICROCHIPS DE
ELECTROFORESI
S
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
MÉTOD
OS

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
MÉTOD
OS

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. CONCLUSION
S
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.