A. Personal Statement 

Since 1997, I have been the associate director of the Center for Biologic Imaging where my efforts have focused on running this state-of-the-art microscopy facility, specifically as the director of the electron microscopy arm of the center. I have also remained highly engaged in any research that is related to tissue and cell imaging, including confocal reconstruction and ultrastructural analysis using transmission and scanning electron microscopy, and freeze-fracture of a variety of organs and cells. As a result of these expertise and collaborations with a variety of principle investigators, I have published over 275 peer-reviewed publications where my contributions have covered all aspects of imaging technologies, including experimental design, execution, interpretation and troubleshooting in light, fluorescence and electron microscopic modalities. Additionally I have been very involved in undergraduate and graduate training. I currently serve as the Director of the Cell Biology and Molecular Physiology Graduate program. I have trained over 30 undergraduates and 3 graduate students in my laboratory and have served on over 40 dissertation committees. I am very involved in classroom, lab and workshop teaching in both the graduate, post-graduate and medical schools in the fields of imaging, general vascular biology, angiogenesis, vasculogenesis, general tissue structure and function with particular concentration in kidney and liver histology. This expertise is helpful in assisting with collaborations where training of students and staff is involved. As such, I am very qualified to assist Dr. Zhou Wang and his laboratory in the imaging aspect of his grant entitled “Luminal Epithelial Junctions, Polarity, and Permeability in BPH Pathogenesis". Below are a set of publications where I have been involved in imaging prostate cancer. 

1. Yates, CC, CR Shepard, DB Stolz, A Wells. Co-culturing human prostate carcinoma cells with hepatocytes lead to increased expression of E-cadherin. Br J Cancer. 96(8):1246-1252. 2007 PMID 17406365 

2. Yates C, CR Shepard, G Papworth, A Dash, D Beer-Stolz, S Tannenbaum, L Griffith, A Wells. Novel three-dimensional organotypic liver bioreactor to directly visualize early events in metastatic progression. Adv Cancer Res 97:225-246. 2007. PMID 17419948 

3. Vyas, AR, ER Hahm, JA Arlotti, S Watkins, D Beer-Stolz, D Desai, S Amin, SV Singh. Chemoprevention of Prostate cancer by D,L-Sulforaphane is augmented by pharmacological inhibition of autophagy. Cancer Res. 73(19):5985-5995. 2013. PMID: 23921360

B. Positions and Honors 

Positions and Employment 

1997--2008 Res. Assistant Professor Dept. of Cell Biology and Physiology, University of Pittsburgh 

1997-present Associate Director, Center for Biologic Imaging. University of Pittsburgh 

2001-present Member Faculty, McGowan Institute for Regenerative Medicine, University of Pittsburgh. 

2007-present Member Faculty University of Pittsburgh Cancer Institute 

2008-2012 Associate Professor, University of Pittsburgh Dept. of Cell Biology; 2o appointment in Pathology. 

2012-present Associate Professor with tenure, University of Pittsburgh Dept. of Cell Biology; 2o appointment in Pathology. 

Other Experience and Professional Memberships 

1991- American Society of Cell Biology 

1997- North American Vascular Biology Organization 

1998- Microscopy Society of America 

1999- American Society for Experimental Pathology 

2011- American Physiological Society 

Grant Reviews: 

2008 (November) Ad hoc P30 Special Emphasis Review Panel, Imaging cores reviewer for ZDK1 GRB-8 M2 1; (2 core grants) NIDDK 

2009 (April 7) Ad hoc P01 reviewer (Imaging cores reviewer for ZDK1 GRB-8 M2 1; 1 grant) NIDDK 

2009 (April 13-14) Ad hoc reviewer for Silvio Conti Digestive Diseases Centers Special Emphasis Panel (5 imaging core grants) NIDDK. 

2009 (October 27-29) NIEHS Ad Hoc Reviewer Superfund Research Training Program. (ZES1-LWJ-M (01) Phone review of 1 grant. 

2010 (March 25-26) Ad hoc reviewer for Silvio Conti Digestive Diseases Centers (6 imaging and core grants) NIDDK. ZDK1 GRB-8-M1 

2011 NIDDK GRB-8 (J1) December 2 –Digestive Diseases Core Centers Meeting (4 imaging core grants). 

2014 (April 3-4) Ad hoc reviewer for Silvio Conti Digestive Diseases Centers (6 imaging and core grants) NIDDK ZDK1-GRB-8-M2 1 

2014 (July) Ad hoc reviewer for S10 Shared Instrumentation grants, Electron Microscopy. S10 ZRG1 CB-D (30). (7 EM grants to review). 

Honors

1986 Commonwealth Scholar, University of Massachusetts, Amherst, MA 

1986 Graduated Summa Cum Laude, University of Massachusetts, Amherst, MA 

1997-2000 American Liver Foundation Liver Scholar Award 

1997 Charlotte Geyer Foundation Cancer Research Grant 

2006 Honorable Mention, Olympus Bioscapes Photomicrograph Award 

2011 2nd and 19th place, Nikon Small World Photomicrograph Awards 

2012 Honorable Mention, Nikon Small World Photomicrograph Award 

2012-2013 Biomedical Graduate Student Association Distinguished Mentor Award, University of Pittsburgh 

C. Contributions to Science 

1. Endothelial cell membrane protein polarity and isolation of vascular-specific proteins and cells in vitro and in vivo. As a graduate student, I developed subcellular isolation techniques to evaluate membrane protein polarity in primary endothelial cell monolayers. The cationic colloidal silica membrane perturbation technique allows for isolation of the apical membrane of cell monolayers from basolateral membranes with high yield and enrichment by ionically coupling cationic silica beads (~50 nm in diameter) to the membrane followed by hypotonic lysis of cells and density centrifugation of the modified membranes. The in vitro techniques were modified to isolate proteins and cells from the vascular components of intact tissues. These in vivo techniques took advantage density modification and/or magnetization of the apical membrane of perfused tissues in order to isolate and enrich apical membranes or cells, respectively, from tissues. These techniques remain relevant in today’s context as they lend themselves to proteomics and also to separating out cell specific signals in tissue. Since cationic colloidal silica is not commercially available, I continue synthesize this reagent and readily supply it to any requesting researcher, as well as provide detailed protocols and troubleshooting for adapting the technique to specific isolation conditions. To date, I have provided the reagent to at least 60 investigators in over 19 countries, and continue to make and share the reagent with anyone requesting it. This technique and my expertise will be available to Dr. Wang should he need to use it. 

a. Stolz DB & BS Jacobson. Examination of transcellular membrane protein polarity of bovine aortic endothelial cells in vitro using the cationic colloidal silica microbead isolation procedure. J. Cell Science 103, 39-51. 1992. PMID 1331135 

b. Stolz DB, G Bannish, BS Jacobson. The role of the cytoskeleton and intercellular junctions on the transcellular membrane Protein Polarity of Bovine Aortic Endothelial Cells In Vitro. J. Cell Science 103, 53-68. 1992. PMID 1429910 

c. Jacobson, B.S., DB Stolz & JE Schnitzer. Identification of endothelial cell surface proteins as potential targets for diagnosis and disease. Nature Medicine. 2(4)482-484. 1996 PMID: 8597963 

d. Stolz DB, MA Ross, HM Salem, W M Mars, GK Michalopoulos, K Enomoto. Cationic Colloidal Silica Membrane Perturbation as a Means of Examining Changes at the Sinusoidal Surface During Liver Regeneration. Am. J. Path. 155:1487-1498. 1999. PMID 10550305; PMCID: PMC1866959. 

e. Ross MA, CM Sander, TB Kleeb, SC Watkins, DB Stolz. Spatiotemporal expression of angiogenesis growth factor receptors during the revascularization of regenerating rat liver. Hepatology. 34:1135-1148. 2001. PMID 11732003 

f. Stolz, DB, MA Ross, A Ikeda, K Tomiyama, T Kaizu, DA Geller, N Murase. Sinusoidal endothelial cell repopulation following ischemia/reperfusion injury in rat liver transplantation. Hepatology, 46:1464-1475. 2007. PMID 17929236; PMCID: PMC:2190086. 

2. Hepatocyte and liver sinusoidal endothelium signaling. As a post-doc and into my years as an independent investigator, I have contributed to our understanding of signaling events that contribute to growth factor and intra- and inter-cellular signaling in hepatocytes and endothelium during liver regeneration as well as other systemic events such as sepsis, hypoxia and ageing. My initial work in liver evaluated in vivo growth factor signaling in liver during liver regeneration following 70% partial hepatectomy. The multiple publications from my work as a post-doc established for the first time that signaling events that trigger liver growth occur very rapidly, often within 5 minutes, following surgical resection. Subsequently, as I assumed the associate directorship of the CBI in 1997, my studies focused on multi-modal imaging approaches of cells, parenchymal tissue and vasculature, including the use of confocal reconstruction, quantitative light, fluorescence, transmission and scanning electron microscopy. Over 90 of my 275 peer-reviewed publications are centered on liver biology or associated tissue. Additionally, due to my extensive publication record on liver sinusoidal endothelium, I am considered an expert in this field of liver physiology. 

a. Stolz DB, WM Mars, BE Petersen, T-H Kim, GK Michalopoulos. Growth factor signal transduction immediately following two-thirds partial hepatectomy in the rat. Cancer Research 59:3954-3960. 1999. PMID 10463591 

b. Wack KE, MA Ross, V Zegarra, SC Watkins, DB Stolz. Sinusoidal ultrastructure evaluated during the 

revascularization of regenerating rat liver. Hepatology 33:363-378. 2001. PMID 11172338 (Cover) 

c. Stolz DB, R Zamora, Y Vodovotz, PA Loughran, Y-M Kim, TR Billiar, RL Simmons, SC Watkins. Peroxisomal localization of inducible nitric oxide synthase in rat hepatocytes. Hepatology. 36:81-93. 2002. PMID 12085352 

d. Khan, Z, GK Michalopoulos, DB Stolz. Peroxisomal localization of hypoxia-inducible factors and HIF regulatory hydroxylases in primary rat hepatocytes exposed to hypoxia-reoxygenation Am J Path 126(4):1251-1269. 2006. PMID 17003483; PMCID: PMC1698853. 

e. Sellaro, TL, AK Ravindra, DB Stolz, SF Badylak. Maintenance of hepatic sinusoidal endothelial cell phenotype in vitro using organ-specific extracellular matrix scaffolds. Tissue Eng 13(9):2301-2310. 2007. PMID 17561801 

f. Hwa, AJ, RC Fry, A Sivaraman, PT So, LD Samson, DB Stolz, LG Griffith. Rat liver sinusoidal endothelial cells survive without exogenous VEGF in 3D perfused co-cultures with hepatocytes. FASEB J 21(10):2564-2579. 2007. PMID 17426068 

g. Straub, AC, KA Clark, MA Ross, AG Chandra. S Li, X Gao, PJ Pagano, DB Stolz, A Barchowsky. Arsenic-stimulated liver sinusoid capillarization in mice requires NADPH oxidase-generated superoxide. J. Clinical Invest. 118(12):3980-3989. 2008. PMID 19033667; PMCID: PMC2582440.

h. Gregg SQ, V Gutiérrez, AR Robinson, T Woodell, A Nakao, MA Ross, GK Michalopoulos, L Rigatti, CE Rothermel, I Kamileri, George Garinis, D Beer Stolz, LJ Niedernhofer. A mouse model of accelerated liver aging due to a defect in DNA repair. Hepatology, 55(2):609-621. 2012. PMID 21953681; PMCID: PMC3250572. 

i. Hang TC, Lauffenburger, LG Griffith and DB Stolz. Lipids promote survival, proliferation, and maintenance of differentiation of rat liver sinusoidal endothelial cells in vitro. Am J Physiol Gastrointestinal Liver Physiol 302(3):G375-388. 2012 PMID 22075778. PMCID: PMC3287397. 

j. El Filali, EE, J Hiralall, HA van Veen, DB Stolz, J Seppen. Human liver endothelial cells, but not macrovascular or microvascular endothelial cells engraft in the mouse liver. Cell Transplant. 22(10):1801-1811. 2012 PMID 23044355 (Cover) 

k. Loughran, PA, DB Stolz, SR Barrick, DS Wheeler, PA Friedman, RA Ruchubinski, SC Watkins, TR Billiar. PEX7 and EPB50 target iNOS to the peroxisome. Nitric Oxide. 31:9-19. 2013. PMID 23474170; PMCID: PMC3642215. 

Complete List of Publications