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Acelerated clinical course of prion
desease in mice compromised in
repair of oxidative DNA damage.
Clara M.O. Jalland, Sylvie L. Benestad, Cecilie Ersdal, Katja
Scheffler, Rajikala Suganthan. Yusaku Nakabeppu, Lars Eide,
Magnar Bjoras, Michael A. Tranulis.
INTRODUCTION:
PRION

Infectious
agent

Are not living
organisms

Acumulates
in infected
tissues

Compossed by a
modify protein

They could
propagate

Tissue damage
and cell death

Misfolded
protein
INTRODUCTION
• PRION REPLICATION:
DNA
DNA
damages
damages
Exogenous
Exogenous
Endogenous
Endogenous

Oxidation of
Oxidation of
bases
bases
Alquilation
Alquilation
of bases
of bases
Hydrolysis
Hydrolysis
of bases
of bases
Mismatch of
Mismatch of
bases
bases
GENERAL OBJECTIVE
OBSERVATE AND REPORT THE PROGRESSION OF
EXPERIMENTAL PRION DISEASE IN MICE WITH
COMPROMISE DNA- REPAIR CAPCITY
MATERIALES Y METODOS
El estudio fue llevado a cabo en concordancia con el
reglamento noruego sobre experimentación animal.

Aprobado por el comité de ética para experimentación con
animales del instituto nacional de veterinaria de Noruega.
12
WILDTYPE

30
RATONES

18
TRANSGENICOS

Criterios de inclusión: ratones hembra que pesaran 20g al
momento de la inoculación.
INOCULO:

Todos los ratones
Todos los ratones
excepto los control
excepto los control
fuero inoculados
fuero inoculados
con RML aislado.
con RML aislado.
DISEÑO EXPERIMENTAL:
Ratones

Anestesiados
e inoculados

Monitoreados

PrPsc yy
PrPsc
GFAP
GFAP

Inmuno
Inmuno
histoquímica
histoquímica

Corte
Corte
longitudinal del
longitudinal del
encéfalo
encéfalo

PrPsc
PrPsc

Western Blot
Western Blot

Congelación del
Congelación del
otro hemisferio
otro hemisferio

Sacrificados

Extracción de
órganos
INMUNOHISTOQUIMICA
Utilizada para
Utilizada para
identificar moléculas
identificar moléculas
especificas en cualquier
especificas en cualquier
clase de tejido, donde
clase de tejido, donde
hay una relación
hay una relación
antígeno- anticuerpo.
antígeno- anticuerpo.
Esto se puede ver al
Esto se puede ver al
microscopio cuando se
microscopio cuando se
utiliza una reacción
utiliza una reacción
colorante.
colorante.
INMUNOHISTOQUIMICA:
WESTERN BLOT:

Las proteínas procedentes de
extractos celulares son separadas por
electroforesis, por medio de la
técnica SDS-PAGE. Tras este proceso
las proteínas se transfieren a un filtro
donde hay anticuerpos que
reaccionan con la proteínas de
interés.
PCR EN TIEMPO REAL:
• Plataforma ideal para el desarrollo de pruebas
moleculares para identificación y
cuantificación de agentes infecciosos de
interés clínico.
• Los procesos de amplificación y detección se
realizan en simultanea.
RESULTADOS
RESULTADOS
RESULTADOS
DISCUSSION
AUTHOR

WHAT DID HE SAY

THEY ARE AGREE? YES OR
NO

(Akhtar, S. et al. 2013 ;Tamguney,
G. et al. 2008).

“Many previous studies have
identified genetic elements that
influence the incubation period of
experimental prion disease”

YES

(Guentchev, M et al, 2002)

The direct contribution of ROS to
the pathogenesis
of prion diseases is somewhat less
explored; however, analysis of
brain areas in humans affected by
CJD showed increased levels of
oxidative DNA damage

YES
DISCUSSION
AUTHOR

WHAT DID HE SAY

(Sandberg, M.K. et al 2011)

“the morphological end-point of
prion disease provides little
information about the rapidity of
clinical deterioration in the toxic
phase of prion disease.”

YES

(Brown, D. R. et al. 1997 ; Brown,
D. R et al. 2002).

“Interestingly, an antioxidant

YES

function has been ascribed to the
cellular prion protein PrPC.”

THEY ARE AGREE? YES OR
NO
CONCLUSIONS
This item is important to note that ROS cause more
damage to specific parts of the body such as the
brain, considering that produces neurodegenerative
diseases, it is important to sensitize the population
on the production of ROS, which although they are
synthesized by the organism inevitably, there are
other situations that stimulate their production and
can be avoided. Is important to note as the PRP may
be a protective factor for susceptibility to oxidative
stress.

It is important with this article to see what the real
effect of the ROS, Prion organism level and mainly
they attack not only generate alterations in DNA but
almost all components of the organism such as
proteins, lipids, in the nucleus, mitochondria.
CONCLUSIONS
•

It is important to do these studies to elaborate
It is important to do these studies to elaborate on the alterations and diseases that can be triggered
on the alterations and diseases that can be
starting with a mutation or alteration alteration
triggered starting with a mutation or in DNA, since diseases are usually of great significance when they
start there since way to cure them and the only we can do is observe their progress
in DNA, is no diseases are usually of great
significance when they start there is no way to
cure them and the only we can do is observe
their progress

This article is useful because ititallows us to
This article is useful because allows us to
recognize the natural mechanisms that make the
recognize the natural mechanisms that make the
body to correct the errors caused by agents such as
body to correct the errors caused by agents such as
ROS in DNA, which in this case is injury escicion
ROS in DNA, which in this case is injury escicion
bases and genes are related in this repair as is
bases and genes are related in this repair as is
MUTYH and OGG1 and the severe impact that may
MUTYH and OGG1 and the severe impact that may
have on the body ififany of these suffers aafailure,
have on the body any of these suffers failure,
usually producing tumor lesions.
usually producing tumor lesions.
MAPA CONCEPTUAL:
MAPA CONCEPTUAL:
Semiario biomol

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Semiario biomol

  • 1. Acelerated clinical course of prion desease in mice compromised in repair of oxidative DNA damage. Clara M.O. Jalland, Sylvie L. Benestad, Cecilie Ersdal, Katja Scheffler, Rajikala Suganthan. Yusaku Nakabeppu, Lars Eide, Magnar Bjoras, Michael A. Tranulis.
  • 2. INTRODUCTION: PRION Infectious agent Are not living organisms Acumulates in infected tissues Compossed by a modify protein They could propagate Tissue damage and cell death Misfolded protein
  • 4. DNA DNA damages damages Exogenous Exogenous Endogenous Endogenous Oxidation of Oxidation of bases bases Alquilation Alquilation of bases of bases Hydrolysis Hydrolysis of bases of bases Mismatch of Mismatch of bases bases
  • 5. GENERAL OBJECTIVE OBSERVATE AND REPORT THE PROGRESSION OF EXPERIMENTAL PRION DISEASE IN MICE WITH COMPROMISE DNA- REPAIR CAPCITY
  • 6. MATERIALES Y METODOS El estudio fue llevado a cabo en concordancia con el reglamento noruego sobre experimentación animal. Aprobado por el comité de ética para experimentación con animales del instituto nacional de veterinaria de Noruega.
  • 7. 12 WILDTYPE 30 RATONES 18 TRANSGENICOS Criterios de inclusión: ratones hembra que pesaran 20g al momento de la inoculación.
  • 8. INOCULO: Todos los ratones Todos los ratones excepto los control excepto los control fuero inoculados fuero inoculados con RML aislado. con RML aislado.
  • 9. DISEÑO EXPERIMENTAL: Ratones Anestesiados e inoculados Monitoreados PrPsc yy PrPsc GFAP GFAP Inmuno Inmuno histoquímica histoquímica Corte Corte longitudinal del longitudinal del encéfalo encéfalo PrPsc PrPsc Western Blot Western Blot Congelación del Congelación del otro hemisferio otro hemisferio Sacrificados Extracción de órganos
  • 10. INMUNOHISTOQUIMICA Utilizada para Utilizada para identificar moléculas identificar moléculas especificas en cualquier especificas en cualquier clase de tejido, donde clase de tejido, donde hay una relación hay una relación antígeno- anticuerpo. antígeno- anticuerpo. Esto se puede ver al Esto se puede ver al microscopio cuando se microscopio cuando se utiliza una reacción utiliza una reacción colorante. colorante.
  • 12. WESTERN BLOT: Las proteínas procedentes de extractos celulares son separadas por electroforesis, por medio de la técnica SDS-PAGE. Tras este proceso las proteínas se transfieren a un filtro donde hay anticuerpos que reaccionan con la proteínas de interés.
  • 13. PCR EN TIEMPO REAL: • Plataforma ideal para el desarrollo de pruebas moleculares para identificación y cuantificación de agentes infecciosos de interés clínico. • Los procesos de amplificación y detección se realizan en simultanea.
  • 14.
  • 18. DISCUSSION AUTHOR WHAT DID HE SAY THEY ARE AGREE? YES OR NO (Akhtar, S. et al. 2013 ;Tamguney, G. et al. 2008). “Many previous studies have identified genetic elements that influence the incubation period of experimental prion disease” YES (Guentchev, M et al, 2002) The direct contribution of ROS to the pathogenesis of prion diseases is somewhat less explored; however, analysis of brain areas in humans affected by CJD showed increased levels of oxidative DNA damage YES
  • 19. DISCUSSION AUTHOR WHAT DID HE SAY (Sandberg, M.K. et al 2011) “the morphological end-point of prion disease provides little information about the rapidity of clinical deterioration in the toxic phase of prion disease.” YES (Brown, D. R. et al. 1997 ; Brown, D. R et al. 2002). “Interestingly, an antioxidant YES function has been ascribed to the cellular prion protein PrPC.” THEY ARE AGREE? YES OR NO
  • 20. CONCLUSIONS This item is important to note that ROS cause more damage to specific parts of the body such as the brain, considering that produces neurodegenerative diseases, it is important to sensitize the population on the production of ROS, which although they are synthesized by the organism inevitably, there are other situations that stimulate their production and can be avoided. Is important to note as the PRP may be a protective factor for susceptibility to oxidative stress. It is important with this article to see what the real effect of the ROS, Prion organism level and mainly they attack not only generate alterations in DNA but almost all components of the organism such as proteins, lipids, in the nucleus, mitochondria.
  • 21. CONCLUSIONS • It is important to do these studies to elaborate It is important to do these studies to elaborate on the alterations and diseases that can be triggered on the alterations and diseases that can be starting with a mutation or alteration alteration triggered starting with a mutation or in DNA, since diseases are usually of great significance when they start there since way to cure them and the only we can do is observe their progress in DNA, is no diseases are usually of great significance when they start there is no way to cure them and the only we can do is observe their progress This article is useful because ititallows us to This article is useful because allows us to recognize the natural mechanisms that make the recognize the natural mechanisms that make the body to correct the errors caused by agents such as body to correct the errors caused by agents such as ROS in DNA, which in this case is injury escicion ROS in DNA, which in this case is injury escicion bases and genes are related in this repair as is bases and genes are related in this repair as is MUTYH and OGG1 and the severe impact that may MUTYH and OGG1 and the severe impact that may have on the body ififany of these suffers aafailure, have on the body any of these suffers failure, usually producing tumor lesions. usually producing tumor lesions.