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Full-Text Articles in Medicine and Health Sciences

Establishing The Purity Of Mononuclear Cell Preparations Using Morphology And Flow Cytometry, Adam Holtz, Bs, Adam D. Toll, Md, Susan W. Burkholder, Md, Alicia M. Carlin, Ba, Linda F. Blumstein, Bs, Kathleen A. Schroeder, Bs, Eric M. Behling, Md, Tina B. Edmonston, Md Oct 2009

Establishing The Purity Of Mononuclear Cell Preparations Using Morphology And Flow Cytometry, Adam Holtz, Bs, Adam D. Toll, Md, Susan W. Burkholder, Md, Alicia M. Carlin, Ba, Linda F. Blumstein, Bs, Kathleen A. Schroeder, Bs, Eric M. Behling, Md, Tina B. Edmonston, Md

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Context: Simple tandem repeat loci are used to track bone marrow engraftment using mononuclear buffy coat cells and T-cells. Poor isolation purity of these subpopulations can result in lower analytical sensitivity of the bone marrow engraftment assay by diluting the cell population in question with other nucleated cells. Validation of the mononuclear cell preparation can be performed by flow cytometry or by counting cell populations on the slide. Conclusions: Our results show that the purity of the Histopaque-1077 mononuclear cell preparation is excellent and that morphology may be sufficient to validate the mononuclear cell isolation method if flow cytometry is …


N-Glycosylation Status Of E-Cadherin Controls Cytoskeletal Dynamics Through The Organization Of Distinct Β-Catenin- And Γ-Catenin-Containing Ajs., Basem T Jamal, Mihai Nita-Lazar, Zhennan Gao, Bakr Amin, Janice Walker, Maria A Kukuruzinska Sep 2009

N-Glycosylation Status Of E-Cadherin Controls Cytoskeletal Dynamics Through The Organization Of Distinct Β-Catenin- And Γ-Catenin-Containing Ajs., Basem T Jamal, Mihai Nita-Lazar, Zhennan Gao, Bakr Amin, Janice Walker, Maria A Kukuruzinska

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

N-glycosylation of E-cadherin has been shown to inhibit cell-cell adhesion. Specifically, our recent studies have provided evidence that the reduction of E-cadherin N-glycosylation promoted the recruitment of stabilizing components, vinculin and serine/threonine protein phosphatase 2A (PP2A), to adherens junctions (AJs) and enhanced the association of AJs with the actin cytoskeleton. Here, we examined the details of how N-glycosylation of E-cadherin affected the molecular organization of AJs and their cytoskeletal interactions. Using the hypoglycosylated E-cadherin variant, V13, we show that V13/β-catenin complexes preferentially interacted with PP2A and with the microtubule motor protein dynein. This correlated with dephosphorylation of the microtubule-associated protein …


Intra-Tumor Heterogeneity Of Mlh1 Promoter Methylation Revealed By Deep Single Molecule Bisulfite Sequencing., Katherine E Varley, David G Mutch, Tina B Edmonston, Paul J Goodfellow, Robi D Mitra Aug 2009

Intra-Tumor Heterogeneity Of Mlh1 Promoter Methylation Revealed By Deep Single Molecule Bisulfite Sequencing., Katherine E Varley, David G Mutch, Tina B Edmonston, Paul J Goodfellow, Robi D Mitra

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

A single tumor may contain cells with different somatic mutations. By characterizing this genetic heterogeneity within tumors, advances have been made in the prognosis, treatment and understanding of tumorigenesis. In contrast, the extent of epigenetic intra-tumor heterogeneity and how it influences tumor biology is under-explored. We have characterized epigenetic heterogeneity within individual tumors using next-generation sequencing. We used deep single molecule bisulfite sequencing and sample-specific DNA barcodes to determine the spectrum of MLH1 promoter methylation across an average of 1000 molecules in each of 33 individual samples in parallel, including endometrial cancer, matched blood and normal endometrium. This first glimpse, …


Decorin Is A Novel Antagonistic Ligand Of The Met Receptor., Silvia Goldoni, Ashley Humphries, Alexander Nyström, Sampurna Sattar, Rick T Owens, David J Mcquillan, Keith Ireton, Renato V Iozzo May 2009

Decorin Is A Novel Antagonistic Ligand Of The Met Receptor., Silvia Goldoni, Ashley Humphries, Alexander Nyström, Sampurna Sattar, Rick T Owens, David J Mcquillan, Keith Ireton, Renato V Iozzo

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Decorin, a member of the small leucine-rich proteoglycan gene family, impedes tumor cell growth by down-regulating the epidermal growth factor receptor. Decorin has a complex binding repertoire, thus, we predicted that decorin would modulate the bioactivity of other tyrosine kinase receptors. We discovered that decorin binds directly and with high affinity (K(d) = approximately 1.5 nM) to Met, the receptor for hepatocyte growth factor (HGF). Binding of decorin to Met is efficiently displaced by HGF and less efficiently by internalin B, a bacterial Met ligand. Interaction of decorin with Met induces transient receptor activation, recruitment of the E3 ubiquitin ligase …


Graft-Versus-Host Disease-Like Pattern In Mycophenolate Mofetil Related Colon Mucosal Injury: Role Of Fish In Establishing The Diagnosis., Kathryn C Behling, Dorothy M J Foster, Tina B Edmonston, Agnieszka K Witkiewicz Jan 2009

Graft-Versus-Host Disease-Like Pattern In Mycophenolate Mofetil Related Colon Mucosal Injury: Role Of Fish In Establishing The Diagnosis., Kathryn C Behling, Dorothy M J Foster, Tina B Edmonston, Agnieszka K Witkiewicz

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

Mycophenolate mofetil (CellCept®), a commonly used immunosuppressive drug in solid organ transplantation, has recently been shown to cause graft-versus-host disease (GVHD)-like changes in the gastrointestinal tract. On rare occasions, true GVHD has also been documented in the gastrointestinal tract of solid organ transplant patients. Because the treatment for these two entities is different, i.e. removal of the offending agent versus the administration of steroids, proper identification of the cause is imperative. We present a case of mycophenolate mofetil colitis mimicking grade I GVHD of the gut. In our study, we used fluorescence in situ hybridization for the Y chromosome to …


Developing Tta Transgenic Rats For Inducible And Reversible Gene Expression., Hongxia Zhou, Cao Huang, Min Yang, Carlisle P Landel, Pedro Yuxing Xia, Yong-Jian Liu, Xu Gang Xia Jan 2009

Developing Tta Transgenic Rats For Inducible And Reversible Gene Expression., Hongxia Zhou, Cao Huang, Min Yang, Carlisle P Landel, Pedro Yuxing Xia, Yong-Jian Liu, Xu Gang Xia

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

To develop transgenic lines for conditional expression of desired genes in rats, we generated several lines of the transgenic rats carrying the tetracycline-controlled transactivator (tTA) gene. Using a vigorous, ubiquitous promoter to drive the tTA transgene, we obtained widespread expression of tTA in various tissues. Expression of tTA was sufficient to strongly activate its reporter gene, but was below the toxicity threshold. We examined the dynamics of Doxycycline (Dox)-regulated gene expression in transgenic rats. In the two transmittable lines, tTA-mediated activation of the reporter gene was fully subject to regulation by Dox. Dox dose-dependently suppressed tTA-activated gene expression. The washout …