- 1. ByCristianFranco CaasSara Correa Prez Plasma proteomic
profile of sulfur mustard exposed lung diseases patients using 2D
gel electrophoresis Molecular Biology2012 Hossein Mehrani1, Mostafa
Ghanei, Jafar Aslani and Zahra Tabatabaei
2. General Information 3. Introduction Sulfur mustard is a
chemical warfare that affect human health Chronic bronchitis, lung
fibrosis, corneal ulcer disease Clinical manifestations in lung
disorders due to sulfur mustard are different Mustard lung is not
responsive to corticosteroids Proteomic can identify and quantify
novel proteins in plasma 4. Introduction Biomarkers of the presence
or severity of disease states Proteomics pattern in bronchoalveolar
lavage (BAL) fluid of SM exposed patients Proteins whose expression
is up or down regulated compared to healthy controls Looking new
markers in the plasma, using ethanol and 2-dimensional gel
Understand nature of long term effects of mustard gas 5.
Proteomic
6. Electrophoresis - 2D gel
- Electrophoresis was defined as the movement of charged
particles in an electric field.
- Its a biochemical separation method analytical, useful for
visualization and separation of protein preparations in small.
- It has the advantage that helps the analyst to determine
quickly how many proteins exist in a particular preparation or
degree of purity of a specific protein preparation.
7. Electrophoresis - 2D gel
- 2-dimensional gel possess properties such as:
- Compatibility with a variety of chemical compounds that makes
an excellent support means for electrophoretic separations.
Disponible en:
http://protocolsonline.com/proteomics/two-dimensional-gel-electrophoresis/
Feb 2012
- Electrophoresis allows the experimental determination of
properties such as isoelectric point and molecular weight (MW) of a
protein.
8. Sulfurd Mustard and Lung Diseases
- Remains a threat to human health.
- Some complications of mustard gas exposure are:
- Interstitial lung diseases
- Chronic obstructive lung disease
- Bronchiolitis obliterans (BO).
Disponible en: http://noticias.lainformacion.com/
politica/armas-biologicas-y-quimicas/fuerzas [on line] Feb 2012 9.
Objective The aim of this study was to identify protein expression
signature or biomarkers that reflect chronic lung damages induced
by SM exposure Disponible en:
http://www.3plusplus.net/2010/11/how-to-objective-missions-in-5th.html
[on -line] Feb 2012 10. Materiales y Mtodos 11. Pacientes
- 20 de sexo masculino con graves condiciones de enfermedad
pulmonar
- 20 de sexo masculino sanos
Inclusin
- Dx, de enfermedad pulmonar crnica debido al gas mostaza.
Exclusin
- Antecedente de una enfermedad crnica
- Tratamiento con corticosteroides(2meses antes)
12. Recoleccin de muestras Tomado de: 1.
http://www.gbo.com/es/index_1611.php
2.http://sobredeporteymas.blogspot.com/, 2.
http://labgeminis.com/plantilla.php?id_sub_seccion=1513-4
http://www.auxilab.es/es/catalogo/vidrio_otros_tubos_Tubo-de-ensayo-borosilicato-20x200-mm.aspx5.
http://www.roche-applied-science.com/sis/complete/index
.jsp?id=com_0200006.
http://laclasedelizpi.blogspot.com/2009/11/para-estudiar-el-clima-de-un-lugar-es.html
[On-line] Febrero 2012 -80C 8-10am 4C->10 min HUPOCombinado de
inhibicin de la proteasa 13. Preparacin de muestras 4 C Disponible
en: 1. http://naturasaludonline.com/2.
http://www.insbal.com/productos.php?categoria=Centrifugas 3.
http://www.um.es/molecula/prot.htm 4.labotienda.com/es/
catalogo/espectrofotometria_espectrofotometros.aspx, 5.
www.directindustry.es/prod/eppendorf/extremos-de-pipetas-22548-449221.htm6.
http://www.gram.rs/ser/ etanol_70.htm [On-line] Febrero 2012 1 2 3
4 5 6 14. Preparacin de muestras 4 C Disponible en: 7.
http://www.infofarma.es/suscripcion_rss/detalle/-/asset_publisher/Ta2g/content/distribucion-comoda-de-agua-ultrapura-para-laboratorios-de-investigacion
/8.
http://www.yarethquimicos.uuuq.com/agitador_vortex_gran_bio_pv-1.htm
9.
http://doc20.comunidadcoomeva.com/blog/.html10.www.directindustry.es/prod/eppendorf/extremos-de-pipetas-22548-449221.htm
12. http://brumasenlanoche.blogspot.com/ On-line] Febrero 2012 7 8
9 10 11 12 15. SDS PageSeparacin las protenas de acuerdo a su
movilidad electrofortica, dependiendo desu carga, forma y tamao.
Electroforesisengeldepoliacrilamidacon dodecil-sulfato sdico
Disponible en: http://en.wikipedia.org/wiki/SDS-PAGE [on-line] Feb
2012 16. ELECTROFORESISENGELDEPOLIACRILAMIDACON DODECILSULFATO
SDICO
- Gel->bajo tamao molecular (30 min2-DEAnlisis del
sobrenadantes y precipitado de etanol recogido. SDS Page
- Tcnica de ionizacin suave utilizada en espectrometra de
masas.
- Anlisis debiomolculasfrgiles
Buffer [ ] Tris-HCl (pH 6,8),2% (w / v) de SDS Azul de
bromofenol0,1% (w / v Glicerol 10% (v / v)Buffer [ ] Urea 7 M
tiourea 1 MTris, (pH 7,5) 20 mM 18. Resultados 19. Figure 1
:SDS-PAGE de protenas plasmticas fraccionadas con diferentes
concentraciones(v/v) de etanol.U,Protenas plasmticas antes de la
precipitacin S, Sobrenadante de etanol P, Precipitado de etanol 20.
Figure 2 Cada gel muestra control saludable;Precipitado de etanol,
(B)Sobrenadante de etanol, (C) Sobrenadante de etanol, Pacientes
con enfermedad pulmonar severa Puntosnumerados indicanlosMADITOFMS
/ MS deprotenas analizadas Patrones2-DEde protenas plasmticas de
pacientes sanos y graves con enfermedades pulmonares expuestos a SM
. 21.
- Each gel shows healthy control;
- (C) severe lung diseases patient; ethanol supernatant
- Numbered spots indicate the MADI TOF MS/MS analysed proteins as
listed in Table 3.
Figure2 22. Discussion 23. Discussion Author Comment Agree Sapey
E. andStockley RA. SerumAmyloid A1is an acute-phase protein that is
induced, like CRP, by inflammatory mediators, including IL-6,
IL-1b, and TNF-a, that rises in acute exacerbation of COPD. Yes
Arredouani et al. Theydescribed the role ofhaptoglobinaffecting the
immune system, showing that haptoglobin directly affects T-cells
and suppress T-helper-cells through down-regulation ofcytokine
production. Yes Mehrani H. et at. In a study of BAL fluid
proteomics patterns in SM exposed patients, thehaptoglobinisoforms
were significantly elevated in moderate and severe lung disease
patients compared to mild and healthy controls Yes Nishiokaet al.
The serum concentration ofhaptoglobinwas decreased during acute
exacerbation in asthmatic children. No 24. Conclusions 25.
Conclussion 26. Conclussion 27. Anexos 28. Plasma proteomic profile
of sulfur mustard exposed lung diseases patients using 2D gel
electrophoresis 29. Plasma proteomic profile of sulfur mustard
exposed lung diseases patients using 2D gel electrophoresis 30.
References 31.
- Mehrani H, Ghanei M, Aslani J, Tabatabaei Z.Plasma proteomic
profile of sulfur mustard exposed lung diseases patients using
2-dimensional gel electrophoresis. Clinical Proteomics. 2011; 8
(2)
- Hernndez G. Mtodos de caracterizacin de protenas:Electroforesis
en gel de poliacrilamid. Disponible en:
http://www.medvet.una.ac.cr/carrera/mva505_Practica3.
pdf.P.35-42
References 32. Thanks Disponible en:
http://suu-construyendounafamilia.blogsp ot.com/2012_01
_01_archive.html [on-line] Feb 2012