SEMINARIO BIO MOLECULAR MANUELA MEJIA LUIS MELO.pdf

Glyucolysis is reduces in dengue virus 2 infected liver cells

1. Manuela Mejía Jaimes
Luis Manuel Melo Henriquez
Third semester
UPB Medicine

2. DENGUE
INTRODUCTION
According to the World Health Organization,
dengue can be defined as a viral infection that
spreads from mosquitoes (Aedes species) to
humans. it is more common in tropical and
subtropical weathers where there is high humidity.
Most people that get infected by dengue, do not
show any symptoms, but the most common ones are
high fever, body ache, acne, general weakness
among others.

3. Also known as Ebdem-
Meyerhoff pathway.
Consists of a set of reactions
that converts 1 molecule of
glucose into two pyruvate
molecules.
It occurs in aerobic and
anaerobic conditions.
Takes place in the cytoplasm.
First stage of the cellular
respiration process.
INTRODUCTION
GLYCOLISIS
10 steps
Energy-requiring fase and energy-releasing fase

4. GENERAL OBJECTIVE
To analize how Glycolysis is
reduced in dengue virus 2
infected liver cells, while
highlighting the importance
of molecular biology in the
research and treatment of
this disease.

5. METHODS
WESTERN BLOT
Technique used to identify the
presence of a specific protein in a
blood or tissue sample based in an
antigen-antibody reaction.
It was used to confirm the
presence of proteins such as STIP1,
GAPDH, Vinculin, E protein, Actin
and SOD1 analized at 12 & 48
hours in mock infection and DENV
2 infection.

6. METHODS
FLOW CYTOMETRY
Technique used to detect a cell
population in a sample in which
other cells predominate.
It offers information on various
parameters of the analyzed cells
and the relationship between
them.
Used in this study to detect and
quantify the fluorescence signal in
DENV 2-infected Hep3B cells after
being treated with metformin.

7. METHODS
Inmunofluorescence assay:
The immunofluorescence assay detects
specific proteins in cells or tissues. The
process involves fixing and permeabilizing
samples, blocking non-specific sites, and
applying a primary antibody targeting the
protein. A secondary antibody, labeled
with a fluorescent dye, is then added.
After washing away excess antibodies, the
samples are examined under a
fluorescence microscope, where the dye
emits light, revealing the location and
distribution of the target protein.
TAKEN FROMhttps://www.sciencedirect.com/topics/nursing-and-health-
professions/immunofluorescence-test

8. METHODS
PCR
Quantitative reverse transcription
polymerase chain reaction, also
called RT-qPCR, is used to detect
and quantify RNA. Total RNA or
mRNA is first transcribed into
complementary DNA (cDNA). The
cDNA is then used as the template
for the quantitative PCR or real-
time PCR reaction (qPCR).
TAKEN FROMhttps://www.nature.com/articles/s41598-023-38383-4

9. RESULTS

10. RESULTS

11. RESULTS

12. RESULTS

13. Author What did the author say?
The author
agrees or not?
Fontaine and colleagues
DENV infection both induces and requires glycolysis for
optimal viral replication, and it was proposed that the
enhancement of glycolysis was required to support the
metabolic requirements for DENV replication.
NO
Allonso and colleagues
DENV NS1 protein directly interacts with GAPDH and
increases glycolytic flux. The expression of DENV NS1
protein alone could increase glycolytic flux.
NO
Silva and colleagues
DENV NS3 protein interacted with GAPDH and reduced
glycolytic activity. Decrease in GAPDH expression, and a
decrease in glycolytic flux as indicated by significant
increase in NADH levels and a significant decrease in the
NAD+/NADH ratio.
YES
DISCUSSION

14. CONCLUSION
DENV infection in the liver leads to significant
proteome changes. Proteomic analysis shows
GAPDH downregulation in DENV-2 infected
Hep3B and iPSC-derived hepatocytes. Elevated
NADH and a lower NAD⁺/NADH ratio suggest
reduced glycolysis, with metformin reducing
infection and virus production. This indicates a
shift to nucleoside production and increased β-
oxidation, highlighting the need for detailed
molecular insights to develop targeted
therapies.

15. CONCEPT MAP

16. CONCEPT MAP