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Prions in the Urine of Patients with Variant Creutzfeldt–Jakob Disease
1. Prions in the Urine of Patients with Variant
Creutzfeldt–Jakob Disease
Laura Quintero
Daniela Márquez Obando
III Semestre
2.
3. INTRODUCTION
Creutzfeldt-Jakob Disease
(CJD)
• Degenerative neurological disorder.
• CJD is caused by an agent called a
prion.
• Symptoms: progressive dementia,
memory loss, personality changes,
hallucinations, myoclonus, ataxia and
changes in gait.
• Epidemiology: occurring in about one
out of every one million people every
year
4. INTRODUCTION
PRIONS (Proteinaceous infectious particles)
• Infectious agent composed of protein in a
misfolded form.
• Pathogenic sialoprotein havyng altered secundary
structure.
• Acts on the same type of protein present in the
body.
• Its transmission has been linked to consumption of
prion-contaminated cattle meat.
6. INTRODUCTION
Relation CJD with Prions
• Prions cause fatal neurodegenerative
disease.
• Prpc PrpSc in prions pathologies.
• Limited proteolysis: Are added within the
central nervous system to form amyloid
plaques.
7. OBJECTIVE
Investigate the presence of
prions in patients with
different transmissible
spongiform encephalopathies,
primarily in Creutzfeldt-Jakob
disease.
9. MATERIALES Y METODOS
Western Blot o Inmunoblot
Fundamento
1. Separación por tamaño de proteínas por electroforesis
en gel (SDS).
2. Transferencia de proteínas a un filtro.
3. Incubación con anticuerpos (reacción con proteínas).
4. Detectar anticuerpo unido al filtro observando el gel.
12. RESULTADOS
DETECCIÓN DE PRPSC EN ORINADE PACIENTES CON LA VARIANTE DE
LA ENFERMEAD DE CREUTZFELD-JAKOB (PMCA)
Solo la muestra control (banda 9) y la muestra de un paciente
con la variante (banda 3) fueron positivos para PRPSC . Las
muestras de otras enfermedades fueron negativas.
13. RESULTADOS
• DETECCIÓN DE PRPSC EN ORINADE PACIENTES CON LA VARIANTE
DE LA ENFERMEAD DE CREUTZFELD-JAKOB (PMCA)
13/14 muestras de pacientes con la variante de la
enfermedad fueron positivos para PRPSC .
La técnica PMCA solo detecta en pacientes esta enfermedad.
14. RESULTADOS
DETECCIÓN DE PRPSC EN ORINADE PACIENTES CON LA VARIANTE DE
LA ENFERMEAD DE CREUTZFELD-JAKOB (PMCA)
Se hicieron experimentos para mirar su las muestras de orina de
diferentes grupos de control eran positivas, sin embargo fueron negativas.
15. RESULTADOS
Después de la digestión con proteinasa K el comportamiento
electroforético de la PRPSC (sin tener en cuenta el país) fue el esperado
16. RESULTADOS
DETECCIÓN DE PRPSC EN ORINADE PACIENTES CON LA VARIANTE DE
LA ENFERMEAD DE CREUTZFELD-JAKOB (PMCA)
Para determinar si la PRPSC es detectable en diferentes momentos de la fase
clínica de la enfermedad se tomaron dos muestras
PRPSC +
Características similares
Mayor cantidad en la última muestra
17. RESULTADOS
CONCENTRACIÓN DE EN ORINA DE PACIENTES CON LA VARIANTE DE
LA ECJ
se llevó a cabo por medio de pmca, su estimación se basó en lo obtenido en el
western blot y las [] de priones recombinantes.
Concentración en cerebro: 100 microgr/gr
Concentración orina: 1 x10-16 gr/ml
18. DISCUSION
AUTHOR COMMENT AGREEMENT OR
DISAGREEMENT
30. Newman PK, Todd NV,
Scoones D,
et al
“The one patient with urine sample that was found to
be negative has a disease duration of 114 months
and was receiving treartment with
intracerebroventricular administration of pentosan
polysulfate, and agent that has been studied for the
treatmen of variant CJD”
disagreement
5. Wadsworth JD, Joiner S, Hill
AF, et al.
31. Brandner S, Whitfield J,
Boone K, et al.
32. Head MW, Ritchie D, Smith N,
et al
This result is not entirely surprising, considering that
variant Creutzfeldt–Jakob disease is acquired by
infection (probably by oral ingestion), and studies
have shown more extensive peripheral involvement
in variant Creutzfeldt–Jakob disease than in the
sporadic or inherited forms of transmissible
spongiform encephalopathies
disagreement
5. Wadsworth JD, Joiner S, Hill
AF, et al.
32. Head MW, Ritchie D, Smith N,
et al
34. Notari S, Moleres FJ, Hunter
SB, et al.
Indeed, PrPSc is readily detectable by means of
immunohistochemical or biochemical studies in
lymph nodes, tonsil, spleen, portions of the intestinal
tract, kidney, liver, pancreas, and skeletal muscle
from patients with variant Creutzfeldt–Jakob disease
disagreement
19. CONCLUSIONS
• This work is a way to promote the investigation, reading and
analysis of various topics related to our career.
• This seminar allows us to apply prior knowledge to
understand this content. With this we can strengthen
important concepts of molecular biology.
• The use of articles in English allow us improve our second
language and to kwon vocabulary about science.
• With this seminar we can to kwon better the molecular biology
techniques and how these techniques works.