Increased levels of prothrombin, apolipoprotein A-IV, and haptoglobin were found in the cerebrospinal fluid of Huntington's disease patients compared to controls. Proteomic studies on cerebrospinal fluid are important for determining if increases in these proteins are correlated with Huntington's disease development and could serve as specific biomarkers. While BBB integrity was maintained, the findings suggest the proteins are produced in the central nervous system and are not related to serum concentrations, indicating a role in the disease itself rather than passive diffusion from blood.

1. “INCREASED PROTHROMBIN, APOLIPOPROTEIN A-IV, AND HAPTOGLOBIN IN THE CEREBROESPIAL FLUID OF PATIENTS WITH HUNTINGTON´S DISEASE.”Yen-Chu Huang.Maria Camila Montufar PantojaHeyly Milena OrtegaIII SEMESTER2011UNIVERSIDAD PONTIFICIA BOLIVARIANA

2. GENERAL OBJETIVEDemonstrate how Prothrombin, Apolipoprotein A-IV and Haptoglobin concentrations in CSF has an important role in the HD evolution and compare these concentrations in HD and control patients .

3. INTRODUCTIONHD is a hereditary, degenerative brain disorder. It is caused by an unstable CAG trinucleotide. Today there are others proteins involved in HD development and symptoms (Apolopoprotein A-IV, Haptoglobin , Prothrombin) and its important to clarify throughproteomicsstudies in CSF wich are theconcentrations (up ordown) and ifthislevels of biomarkers are correlated in HD.

4. PROTHROMBINPlasma protein synthesized in the liver.Itplaysanimportantfunctionin coagulation processby reaction with the thromboplastin enzyme , with Ca ++ cation and V activate factor as coenzymes.In HD prothrombin proinflammatoryagent : DISEASE STATUS BIOMARKERXaCa++VaTromboplastina

5. APOLIPOPROTEIN AIV (APO AIV)glycoprotein synthesized by the human intestine and liver The formation of chylomicrons acts as a signal for the induction of Apo AIV synthesisalters the activity of the enzymes (LPL and LCAT) of lipoprotein metabolism and cholesterol efflux from extrahepatic tissuesIs involved in the long-term regulation of both food intake and body weight

6. HAPTOGLOBINprotein produced by the liverhaptoglobin binds free hemoglobin (Hb) released from erythrocytes with high affinity and thereby inhibits its oxidative activityThe haptoglobin-hemoglobin complex will then be removed by the reticuloendothelial system It prevents loss of iron through the kidneys and protecting the kidneys from damage by hemoglobin

7. ENFERMEDAD DE HUNITNGTON - HDProgressive neurodegenerative disorder with autosomaldominant inheritance: - MovementDisorders.- Cognitiveimpairment.- Psychiatricdisorders.

8. MATERIALES Y METODOSPREPARACION DE LAS MUESTRASLCR y suero extraido de 9 HD ( 5 hombres- 4 mujeres).LCR y suero de 18 pacientes tipo control (7 hombres- 11 mujeres). Cuatro grupos:1 Electroforesis de dos dimensiones.

9. 3 Cuantificacion de protrombina, Apo-A IV y haptoglobina.

10. ELECTROFORESIS DE DOS DIMENSIONES (2-DE ) Técnica de alta resolución cuyo objetivo es la separación de mezclas de proteínas altamente complejas. las proteínas son separadas por dos criterios físicos. Punto isoeléctrico (isoelectroenfoque).

11. Masa molecular por electroforesis discontinua en gel de poliacrilamida en presencia de SDS (SDS-PAGE). DIGESTION EL GELForma parte de la preparación de las muestras para la espectrometría de masas, consta de cuatro pasos: Decoloración.

12. Reducción y alquilación.

13. Corte de la proteína serina proteasa tripsina

14. Extracción del péptido generado.ESPECTROMETRIA DE MASAtécnica experimental que permite la medición de iones derivados de moléculasse basa en el fenómeno conocido como desbastado (sputtering, en inglés) de partículas centradas en un blanco, que son bombardeadas por iones, átomos o moléculas. Dependiendo del intervalo de energía de la partícula primaria, ocurren colisiones.el patrón de fragmentación resultante así como losiones residuales constituyen el espectro de masas

15. WESTERN BLOTBusco proteínas especificasUso acUso técnica SDS-PAGESeparación de proteínas por electroforesis en gel

16. ELISAEnsayo por inmunoabsorción ligado a enzimasFosfatasa alcalina—peroxidasa de rabanoDetección ag-ac3 tipos:Ensayo tipo sandwichEnsayo competitivoEnsayo indirecto

17. RESULTADOSRepresentative Western blots of prothrombin (72 kD) and albumin (64 kD) in CSF and serum of HD patients and their correspondingcontrols.

18. RESULTADOSRepresentative Western blots of Apo A-IV (46 kD) and albumin (64 kD) in CSF and serum, respectively, of HD patients and theircorrespondingcontrols (C).

19. RESULTADOSQalb, CSF haptoglobin, and serumhaptoglobinconcentration in HD patients.

20. DISCUSSION

21. CONCLUTIONSTheproteomicstudies in CFS in patients HD and control patients are veryimportanttodeterminateiftheincreaselevels of Haptoglobin, Apolipoprotein A-IV and Prothrombin are correlatedwith HD development.Itisimportanttorecognizethisproteinslikespecificsbiomarkers in HD; thatswhytheconcentrations of eachonewerequantified in CSF and serum; thusindirectlyevaluatethe BBB integrity.

22. CONCLUTIONSTo be able to correlate the levels of these proteins with other mental diseases such as Alzheimer's, multiple sclerosis, among others; allow us to determine whether the activity of proteins mentioned in the study are a common phenomenon in a variety of neurodegenerative-tive disease or are specifically for HD.Becouseof the intact values of BBB and the increase levels of proteins, it suggest that they are produced in the CNS itself and that the relation within de concentrations of proteins in CSF has nothing to do with the concentration of these in serum.

23. MAPA CONCEPTUAL

24. MAPA CONCEPTUAL

25. BIBLIOGRAFIAMARTINEZ, L. Biología Molecular. ED UPB. Segunda edición. Medellín. 2011.HUANG Y-C, TSENG M-Y, et al. Increasedprothrombin, Apolipoprotein A-IV, and Haptoglobin in thecerebrospinal fluid of patientswithHuntington`sdisease. Enero 31, 2011