Enrico Bertini, convegno Mitocon 2013

803 views

Published on

il protocollo diagnostico per bambini
Enrico Bertini, O.P. Bambino Gesù, Roma

Published in: Technology, Health & Medicine
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
803
On SlideShare
0
From Embeds
0
Number of Embeds
313
Actions
Shares
0
Downloads
0
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

Enrico Bertini, convegno Mitocon 2013

  1. 1. Il protocollo diagnostico per bambini: esperienze a confrontoEnrico Bertini – O.P. Bambin Gesù Roma
  2. 2. Diagnosi clinica• Importanza di stabilire una diagnosi sindromica(clinica, biochimica clinica, MRI, biopsia muscolare, biochimica tissutale)• S. di Leigh, MELAS, MERRF, KSS, Leucodistrofia, PCH, epatopatia progressiva,anemia sideroblastica, miopatia, neuro-miopatia, epilessia, etc.• Acidosi lattica nel sangue; acidi organici; acidosi lattica liquorale;acidosi lattica in MRS• Revisione critica delle acidosi lattiche neonatali• acidosi lattiche persistente nel tempo?• acidosi lattica si associa a grave scompenso cardiocircolatorio?• da cardiomiopatia? da malformazione cardiaca?• acidosi lattica si associa ad una sepsi?• acidosi lattica si associa a neutropenia, ad una aciduria organica?
  3. 3. Biochimica clinica• Fumaric aciduria (Fumarase deficiency)• Other Krebs cycle Krebs intermediate metabolites (no discrimination)• 3-methylglutaconic aciduria (BARTH syndrome, OPA3-Costeff syndrome)• Mild methylmalonic aciduria (SUCLA2)• alpha 2-ketoglutaric aciduria, oxoglutarate (PDH)• Glutaric aciduria type II (glutaric acid, lactic, ethylmalonic, butyric,isobutyric, 2-methyl-butyric, and isovaleric acids) ETFA, ETFB, ETFDH
  4. 4. MRI
  5. 5. MRIPCL: Pt 2 SCFemaleSib/Pt1Ages 3, 5&7 yrs199419961991
  6. 6. MRI
  7. 7. Biopsia muscolarelTG TGTGTGSDH SDH SDH SDH
  8. 8. Biopsia muscolare
  9. 9. Biopsia muscolare
  10. 10. Biopsia muscolareCOX COX SDH
  11. 11. Biochimica TissutaleSpectrophotometric analysis of muscle (fibro) homogenate•Complex I•Complex II•Complex III•Complex II+III•Complex IV (cytochrome c oxydase)• Cytrate synthaseNative blue electrophoresis and in situ activity (muscle, fibro)•Complex I•Complex II•Complex III•Complex IV (cytochrome c oxydase)•Complex VATPsynthesis on isolated mitochondria from fibroblasts•Complex I•Complex II•Complex VHPLC: Coenzyme Q10
  12. 12. Biochimica TissutaleMUSCLE BRAINC IC IVCP P CBN-PAGE+IN SITU ACTIVITY70mg of mitochondria were submitted to colorimetric enzymatic staining immediatelyafter the electrophoretic run and complex I (CI=NADH dehydrogenase) and complex IV(CIV=cytochrome c oxidase) activities were evaluated as already reported (Zerbetto,Dabbeni-Sala, 1997, Electrophoresis). C= control; P=patient
  13. 13. Biochimica Tissutale0510152025303540SUCCINATE MALATECPCOMPLEX-V ACTIVITYSuccinateMalate
  14. 14. Genetica MolecolareMitoChipExome sequencingSanger sequencing2 Chip• DNA mitocondriale• POLG1, Twinkle, PDH (E1alfa), DGOUK, MPVT17, TK2SUCL2, SUCLG1, RRM2B,Calvo et al. Molecular diagnosis of infantile mitochondrial disease with targetednext-generation sequencing. Sci Transl Med. 2012 Jan 25;4(118):118ra10.

×