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Proposal1

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  • Researches as Malformations of cortical development in children with congenital cytomegalovirus infection - A study of nine children with proven congenital cytomegalovirus infection affirm that... “Congenital cytomegalovirus (CMV) infection is the most common vertically transmitted disease with the rate of the infection ranging from 0.2 to 2.4% in newborn infants”.
  • A chemical remedy had already been experimented inOral Hexadecyloxypropyl-Cidofovir Therapy in Pregnant Guinea Pigs Improves Outcome in the Congenital Model of Cytomegalovirus Infectionas an alternative orally supply analog of the existing drug, Cidofovir, against CMV
  • 125 pregnant women tested, IgG antibody was found 84% (105 women) of the times, but IgM only on 7.2% (9 women)It means that they weren’t always present, signifying lack of antibody levels. Therefore, that can be reasoned that... (hypothesis)
  • Musmusculus, because:resemblance to human, able to be infected with CMV, more ethical to work with than humans, small sized, highly fertile, principally because isgraded as ideal for ‘in vivo’ hybridomasto produce continuous supply of antibody. Division:9 for each experimental group + 3 for the control group.Image Citometri = stores image data and the microscopic parameters describing the optics during the acquisition.
  • Dextran Leukocyte Separation Kit = isolation of the PMN by dextran separation PMN Polymorphonuclear leukocytes:CMV DNA can be detected in different fractions of leukocytes like this one during active infectionFormaldehyde-Noidet P-40=solution made of a simple Aldehyde and a detergent.MRI= Magnetic Resonance Image
  • Selective Optical Control of Synaptic Transmission in the Subcortical Visual Pathway by Activation of Viral Vector-Expressed Halorhodopsin.Anestethiaze with ketamineand xylazineInject mice with unilateral pressure injections of 1.0–1.5 µl of CMV vector solution (Recombinant CMV) into the body using a glass micropipette with a tip diameter of 20–30 µm.Place pipette for an 30 s and then slowly withdrawn.  
  • IL 6 =B-cell stimulatory factorIL 28=inmune defense against virusIgG=  battle bacterial and viral infectionsIgM= first reponse against antigen presenceEach will be for a complete of period fifteen days, because the sampling can be made days before the offspring born.
  • After fifteen days of gestation, the CVS (Chronic Villous Sampling) in order to analyze the placenta tissue. Placenta is the organ where the mother supplies the fetus important needs for the development as nutritional and respiratory.
  • Transcript

    • 1. Increase antibody production as treatment against development of Lissencephaly caused by CMV in pregnant Musmusculus
      Celizbets Colón Ortiz
      RISE (Summer Bridge)
      July 12, 2011
    • 2. Background
    • 3. Lyssencephaly
      Smoothbrain (froomtheGreek“lissos” and “enkhapalos”)
      Cerebral cortexscarces of:
      Gyri
      Groove
      Fissures
    • 4. Malformation of thephysicalstructureisbecauseitdidn’tdevelopedcompletelywhileembryonicdevelopment.
      Lack of embryonicdevelopment:
      Low blood flow
      CongenicCitomegalovirus (cCMV)
      Genetic mutation (LIS1, XLIS)
    • 5. Researches
      Malformations of cortical development in children with congenital cytomegalovirus infection - A study of nine children with proven congenital cytomegalovirus infection
      Development of Lissencephaly in the fetus as direct consequence of early infection.
      Most common vertically transmitted disease
      rate of the infection ranging:
      0.2 to 2.4% in newborn infants
      40.000 infants per year
    • 6. Chemical remedy
      Oral Hexadecyloxypropyl-Cidofovir Therapy in Pregnant Guinea Pigs Improves Outcome in the Congenital Model of Cytomegalovirus Infection
      Alternative orally supply analog of the existing drug, Cidofovir
      Only proved effiency of Guinea Pigs
    • 7. Why supply antibodies?
    • 8. Hypothesis
      The augment of antibodies IgG and IgM may reduce the presence of CVM, representing a minor risk to the fetus in developing Lissencephaly.
    • 9. Objectives
      • Develop a treatment with antibodies against the growth of Citomegalovirus that causes Lyssencephaly
      • 10. Reduce the CMV presence in the placenta
      • 11. Lower the risks of developing Lissencephaly caused by cCMV.
    • Methodology
    • 12. Materials
      CMV Pp38 (UL80a), Cytomegalovirus Antigen, Recombinant
      IgMAntibody, mAb, Mouse
      Mouse IgG control (Whole Molecule), Purified
      IL-4 Antibody (2G6A8), mAb, Mouse
      Mouse IL-28A/B
      39 female pregnant MusMusculus
      Image Cytometri
    • 13. Light diagnostics CMV direct Immunofluorescence Assay Kit
      Dextran Leukocyte Separation Kit
      Formaldehyde-Noidet P-40
      MRI
    • 14. Methodology
    • 15. Step: Prepare research models
    • 16. Prepare research models
      Mice from 12-13 days
      Anesthiaze mice
      Inject with CMV vector solution with a glass micropipette
      Place pipette for an 30 s and then slowly withdrawn.
    • 17. Research model’s distribution
      Groups:
      IL 6
      IL 28
      IgG
      IgM
      Sub-groups:
      A= every week
      B= every 48 hrs
      C= every three days
    • 18. Step: Viral presence assays
    • 19. Extract serology: CVS
      Placenta is the common path
      Chronic Villous Sampling
      Suction of a tissue from placenta
    • 20. Step: ELISPOT
      Quantify presence of IgG and IgM in all experimental groups
      Procedure according to:
      “General ELISPOT procedure” by abcam.com
    • 21. Step: Direct immunofluorescence
      Cualitative presence of IgG and IgM in all experimental groups
      Process according to:
      “Immunostaining of cells in tissue culture” by ENCORE biotechnology INC.
    • 22. Spected results
      ELISPOT will indicate:
      augment in antibodies in experimental groups
      will mean lower presence of CMV
      Direct immunofluorescence:
      Control group will be brighter than experimental groups
      will mean high presence of CMV
    • 23. Bibliography
      Bosnjak VM, Daković I, et al. Malformations of cortical development in children with congenital cytomegalovirus infection - A study of nine children with proven congenital cytomegalovirus infection. January, 2011. http://www.ncbi.nlm.nih.gov/pubmed/21648339
      Ravo FJ, Bernstein DI, et al. Oral Hexadecyloxypropyl-Cidofovir Therapy in Pregnant Guinea Pigs Improves Outcome in the Congenital Model of Cytomegalovirus Infection. November 15, 2011. http://www.ncbi.nlm.nih.gov/pubmed?term=Oral%20Hexadecyloxypropyl-Cidofovir%20Therapy%20in%20Pregnant%20Guinea%20Pigs%20Improves%20Outcome%20in%20the%20Congenital%20Model%20of%20Cytomegalovirus%20Infection%20
    • 24. Araswathy TS, Az-Ulhusna A, et al. Seroprevalence of cytomegalovirus infection in pregnant women and associated role in obstetric complications: a preliminary study. March, 2011. http://www.ncbi.nlm.nih.gov/pubmed/21710852
      Kaneda K, Kasahara H, et al.Selective Optical Control of Synaptic Transmission in the Subcortical Visual Pathway by Activation of Viral Vector-Expressed Halorhodopsin. April 5, 2011. http://www.ncbi.nlm.nih.gov/pubmed?term=Selective%20Optical%20Control%20of%20Synaptic%20Transmission%20in%20the%20Subcortical%20Visual%20Pathway%20by%20Activation%20of%20Viral%20Vector-Expressed%20Halorhodopsin
      Boeckh, M. and Boivin, G. Quantitation of Cytomegalovirus: Methodologic Aspects and Clinical Applications. Clinical Microbiology Reviews, July 1998, p. 533-554, Vol. 11, No. 3. http://cmr.asm.org/cgi/content/full/11/3/533