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Laura Quintero
Daniela Márquez Obando
III Semestre
Prions in the Urine of Patients with Variant Creutzfeldt–Jakob Disease
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
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.
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.
OBJECTIVE
Investigate the presence of prions in patients with different transmissible spongiform encephalopathies, primarily in Creutzfeldt-Jakob disease.
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.
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.
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.
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.
RESULTADOS
Después de la digestión con proteinasa K el comportamiento electroforético de la PRPSC (sin tener en cuenta el país) fue el esperado
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
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
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
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.