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Jung-Hyun Park, Yoon Mi Jin, Soojin Hwang, Du-Hyong
Cho, Duk-Hee Kang, Inho Jo
Carolina Mora Sánchez
Laura María Moreno León
Are a group of disorders of
the heart and blood vessels
Are the number one
cause of death globally
Can be prevented by
addressing risk factors.
>Coronary heart disease.
>Cerebrovascular disease.
Seminario molecular..! cms & lml
Seminario molecular..! cms & lml
Most uric acid dissolves in blood
and travels to the kidneys, where
it passes out in urine. We can use
uric acid levels in the diagnosis
different pathologies
Is the end product of purine
metabolism in humans
In normouricemic
humans, the daily
production and
ingestion of uric acid
is well balanced by
the daily excretion
hyperuricemia is associated with hypertension
cardiovascular diseases.
eNOS is responsible
for most of the
vascular NO
produced.
That increased blood flow
by dilating blood vessels
Is a gas and is an important
protective molecule in the
vasculature
Dysregulation contributes
to the pathogenesis of
certain diseases Dimerization.
Interaction with calmodulin
(CaM) and
tetrahydrobiopterin (BH4),
Acetylation
Phosphorylation.
eNOS is regulated by
Mediates the control of a large
number of enzymes, ion channels
and other proteins by Ca2+.
Is a calcium-binding
messenger protein
expressed in all
eukaryotic cells.
Among the enzymes to be stimulated by
the calmodulin-Ca2+ complex are a number
of protein kinases and phosphatases.
Bovine aortic endothelial cells (BAEC)
Human umbilical vein EC (HUVEC)
Human plasma and aortic EC – in vitro
Pulmonary artery EC
eNOS CaM in EC
eNOS activity
NO production
Reducing the
binding between
Explain how uric acid
decreases NO production
by direct modulations on
eNOS, leading to causing
cardiovascular problems.
Ácido úrico
eONS bovina recombinante
eNOS, p-eNOS-Thr 495, p-eNOS-Ser1177, p-eNOS- Ser114
Calmodulina recombinanteFluo-4 AM
Ig CaM
FBS
M200Ig Lisina acetilada
LSGSDPBS NCSL-glutamina
Penicilina y estreptomicina
MEM
Tripsina EDTA
Método para el estudio del
comportamiento de las células, muy
adecuado como modelo para el estudio
del desarrollo y la diferenciación, por lo
que las líneas celulares que conservan
la capacidad de diferenciarse
La producción de ON es medida como concentración de
nitrito en la células cultivadas.
La cuantificación de la actividad de eNOS se
lleva a cabo midiendo la conversión de L-arginina
en L citrulina, en presencia de concentraciones
saturantes de los cofactores enzimáticos
adecuados.
Seminario molecular..! cms & lml
Proteínas procedentes de extractos celulares son
separados por electroforesis en gel según su tamaño kDa
Técnica: SDS-PAGE policrámida en
la que son disueltas en una solución,
con el detergente sodio duodecil
sulfato.
Manipulación que se hace con
células o sistemas aislados de
células, en el laboratorio con los que
se trata de simular las condiciones
que tendrían en los organismo de los
que fueron aislados
Es una técnica popular para identificar
las interacciones proteína-proteína
fisiológicamente relevantes mediante el
uso de anticuerpos para determinar la
presencia de una proteína específica.
Seminario molecular..! cms & lml
Es una enzima del ciclo de la urea que
cataliza la hidrólisis de la arginina,
produciendo ornitina y urea.
Amplificación de RNA a través de la
síntesis previa de su cDNA (DNA copia),
utilizando una transcriptasa inversa,
seguida de varios ciclos de PCR
convencional.
Determinar
expresión genética
en diversos tejidos.
Seminario molecular..! cms & lml
Seminario molecular..! cms & lml
Seminario molecular..! cms & lml
Seminario molecular..! cms & lml
Seminario molecular..! cms & lml
RESEARCHER COMMENT AGREE DISAGREE
D.I Feing
¨Epidemiology
studies showed
that higher blood
level of uric
acid is significantly
associated with
the risk of
cardiovascular
diseases¨.
X
RESEARCH
ER
COMMENT AGREE DISAGREE
I.
Papeziková
¨Most recent
study revealed
that uric acid
decreased NO
production, both
indirectly
and directly; by
increasing
arginase activity
and decreasing
eNOS-Ser1177
phosphorylation,
respectively¨.
X
RESEARCHER COMMENT AGREE DISAGREE
C. Nathan
¨CaM is one of
the key proteins
for eNOS
activation.
Although
CaM binds
tightly to eNOS,
even at the
resting basal
levels of
intracellular
Ca2+ levels¨.
X
RESEARCHER COMMENT AGREE DISAGREE
J. White
¨increased
intracellular Ca2+
by
extrastimuli such
as ionomycin
induced more
association of
CaM
with eNOS¨.
X
• Uric acid inhibits the production of nitric oxide which is
a vasodilator important for the prevention of heart
disease
• This project can help people to understand how if they
do not have healthy habits can cause cardiovascular
disease.
• Ionomycin is used in research to raise the intracellular
level of calcium, this help the interaction between
eNOs y CaM.
• Uric acid reduced the interaction between CaM and
eNOS this produces a disminusion of the production of
NO.
Seminario molecular..! cms & lml
Carolina Mora Sánchez

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Seminario molecular..! cms & lml

  • 1. Jung-Hyun Park, Yoon Mi Jin, Soojin Hwang, Du-Hyong Cho, Duk-Hee Kang, Inho Jo Carolina Mora Sánchez Laura María Moreno León
  • 2. Are a group of disorders of the heart and blood vessels Are the number one cause of death globally Can be prevented by addressing risk factors. >Coronary heart disease. >Cerebrovascular disease.
  • 5. Most uric acid dissolves in blood and travels to the kidneys, where it passes out in urine. We can use uric acid levels in the diagnosis different pathologies Is the end product of purine metabolism in humans In normouricemic humans, the daily production and ingestion of uric acid is well balanced by the daily excretion
  • 6. hyperuricemia is associated with hypertension cardiovascular diseases.
  • 7. eNOS is responsible for most of the vascular NO produced. That increased blood flow by dilating blood vessels Is a gas and is an important protective molecule in the vasculature
  • 8. Dysregulation contributes to the pathogenesis of certain diseases Dimerization. Interaction with calmodulin (CaM) and tetrahydrobiopterin (BH4), Acetylation Phosphorylation. eNOS is regulated by
  • 9. Mediates the control of a large number of enzymes, ion channels and other proteins by Ca2+. Is a calcium-binding messenger protein expressed in all eukaryotic cells. Among the enzymes to be stimulated by the calmodulin-Ca2+ complex are a number of protein kinases and phosphatases.
  • 10. Bovine aortic endothelial cells (BAEC) Human umbilical vein EC (HUVEC) Human plasma and aortic EC – in vitro Pulmonary artery EC
  • 11. eNOS CaM in EC eNOS activity NO production Reducing the binding between
  • 12. Explain how uric acid decreases NO production by direct modulations on eNOS, leading to causing cardiovascular problems.
  • 13. Ácido úrico eONS bovina recombinante eNOS, p-eNOS-Thr 495, p-eNOS-Ser1177, p-eNOS- Ser114 Calmodulina recombinanteFluo-4 AM Ig CaM FBS M200Ig Lisina acetilada LSGSDPBS NCSL-glutamina Penicilina y estreptomicina MEM Tripsina EDTA
  • 14. Método para el estudio del comportamiento de las células, muy adecuado como modelo para el estudio del desarrollo y la diferenciación, por lo que las líneas celulares que conservan la capacidad de diferenciarse
  • 15. La producción de ON es medida como concentración de nitrito en la células cultivadas.
  • 16. La cuantificación de la actividad de eNOS se lleva a cabo midiendo la conversión de L-arginina en L citrulina, en presencia de concentraciones saturantes de los cofactores enzimáticos adecuados.
  • 18. Proteínas procedentes de extractos celulares son separados por electroforesis en gel según su tamaño kDa Técnica: SDS-PAGE policrámida en la que son disueltas en una solución, con el detergente sodio duodecil sulfato.
  • 19. Manipulación que se hace con células o sistemas aislados de células, en el laboratorio con los que se trata de simular las condiciones que tendrían en los organismo de los que fueron aislados
  • 20. Es una técnica popular para identificar las interacciones proteína-proteína fisiológicamente relevantes mediante el uso de anticuerpos para determinar la presencia de una proteína específica.
  • 22. Es una enzima del ciclo de la urea que cataliza la hidrólisis de la arginina, produciendo ornitina y urea.
  • 23. Amplificación de RNA a través de la síntesis previa de su cDNA (DNA copia), utilizando una transcriptasa inversa, seguida de varios ciclos de PCR convencional. Determinar expresión genética en diversos tejidos.
  • 29. RESEARCHER COMMENT AGREE DISAGREE D.I Feing ¨Epidemiology studies showed that higher blood level of uric acid is significantly associated with the risk of cardiovascular diseases¨. X
  • 30. RESEARCH ER COMMENT AGREE DISAGREE I. Papeziková ¨Most recent study revealed that uric acid decreased NO production, both indirectly and directly; by increasing arginase activity and decreasing eNOS-Ser1177 phosphorylation, respectively¨. X
  • 31. RESEARCHER COMMENT AGREE DISAGREE C. Nathan ¨CaM is one of the key proteins for eNOS activation. Although CaM binds tightly to eNOS, even at the resting basal levels of intracellular Ca2+ levels¨. X
  • 32. RESEARCHER COMMENT AGREE DISAGREE J. White ¨increased intracellular Ca2+ by extrastimuli such as ionomycin induced more association of CaM with eNOS¨. X
  • 33. • Uric acid inhibits the production of nitric oxide which is a vasodilator important for the prevention of heart disease • This project can help people to understand how if they do not have healthy habits can cause cardiovascular disease.
  • 34. • Ionomycin is used in research to raise the intracellular level of calcium, this help the interaction between eNOs y CaM. • Uric acid reduced the interaction between CaM and eNOS this produces a disminusion of the production of NO.