Mark M Bouzyk

Publications (2)8.04 Total impact

• Source

  Available from: emory.edu

  Article: DNA extraction from formalin-fixed, paraffin-embedded tissue.

  Weining Tang, Freedom B David, Malania M Wilson, Benjamin G Barwick, Brian R Leyland-Jones, Mark M Bouzyk
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  ABSTRACT: Introduction Formalin-fixed, paraffin-embedded (FFPE) tissue is one of the most widely practiced methods for clinical sample preservation and archiving. It is estimated that, worldwide, over a billion tissue samples, most of them FFPE, are being stored in numerous hospitals, tissue banks, and research laboratories. These archived samples could potentially provide a wealth of information in retrospective molecular studies of diseased tissues. While standard for histopathology and microscopic investigation (e.g., hematoxylin and eosin [H&E] staining, immunohistochemistry [IHC], and tissue microarray [TMA]), FFPE samples pose a major challenge for molecular pathologists, because nucleic acids are heavily modified and trapped by extensive protein-nucleic acid and protein-protein cross linking. Historically, FFPE samples were not considered to be a viable source for molecular analyses. Recently, however, it has been discovered that with appropriate protease digestion, it is possible to release microgram amounts of DNA and RNA from FFPE samples. The purified nucleic acids, although highly fragmented, are suitable for a variety of downstream genomic and gene expression analyses, such as polymerase chain reaction (PCR), quantitative reverse transcription PCR (qRT-PCR), microarray, array comparative genomic hybridization (CGH), microRNA, and methylation profiling. Several commercial kits are currently available for FFPE extraction. The protocol reported here is adapted from the Ambion RecoverAll Total Nucleic Acid Isolation Kit, but includes several modifications. Although our protocol focuses on DNA isolation, the RecoverAll Kit can also be utilized to recover RNA, including microRNA.
  Cold Spring Harbor Protocols 02/2009; 2009(2):pdb.prot5138. · 4.63 Impact Factor
• Article: The endothelin-1 G5665T polymorphism impacts transplant-free survival for single ventricle patients.

  Paul M Kirshbom, William T Mahle, Ronald W Joyner, Traci Leong, Malania Wilson, Brian E Kogon, Kirk R Kanter, Mark M Bouzyk
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  ABSTRACT: Early survival for children with single-ventricle congenital heart disease has improved, but late attrition remains a serious problem. The endothelin-1 G5665T single nucleotide polymorphism has been linked to increased vascular reactivity and hypertension. The goal of this study was to determine whether this single nucleotide polymorphism alters transplant-free survival for children with single-ventricle congenital heart disease. DNA was isolated from 165 children with single-ventricle congenital heart disease born between January 1980 and December 2006. The endothelin-1 G5665T single nucleotide polymorphism genotype was determined by using real-time polymerase chain reaction. Kaplan-Meier survival curves were generated with a combined end point of death or transplantation. The Cox proportional hazard method was used to evaluate potential covariates. The endothelin-1 G5665T genotype was significantly associated with transplant-free survival for the group as a whole (P = .002), with the greatest effect in children with hypoplastic left heart syndrome (n = 64, P = .0002) as opposed to patients with other types of single-ventricle anatomy (n = 101, P = .1). Cox proportional hazard modeling revealed 3 independent predictors of poor outcome: endothelin G5665T genotype (T/T genotype, P = .001), prematurity (gestational age <37 weeks, P = .02), and era of surgical intervention (1990s vs other decades, P = .02). Long-term survival of single-ventricle patients is dependent on many factors. These data suggest that genetic variability involving vascular resistance modifiers, such as endothelin-1, might play an important role, particularly in patients with hypoplastic left heart syndrome. Future studies should include evaluation of the relationship between endothelin genotype and plasma endothelin levels and possibly therapeutic trials of endothelin blockers in high-risk patients.
  The Journal of thoracic and cardiovascular surgery 07/2008; 136(1):117-22. · 3.41 Impact Factor