University of Pittsburgh Cancer Institute

3 ARTICLES PUBLISHED IN JoVE

Biology

Quantitative, Real-time Analysis of Base Excision Repair Activity in Cell Lysates Utilizing Lesion-specific Molecular Beacons

David Svilar^1,2, Conchita Vens³, Robert W. Sobol^1,2,4

¹Department of Pharmacology & Chemical Biology, University of Pittsburgh School of Medicine, ²Hillman Cancer Center, University of Pittsburgh Cancer Institute, ³Department of Experimental Therapy, The Netherlands Cancer Institute, ⁴Department of Human Genetics, University of Pittsburgh School of Public Health

We describe a method for the quantitative, real-time measurement of DNA glycosylase and AP endonuclease activities in cell nuclear lysates. The assay yields rates of DNA Repair activity amenable to kinetic analysis and is adaptable for quantification of DNA Repair activity in tissue and tumor lysates or with purified proteins.

Cancer Research

Modified Terminal Restriction Fragment Analysis for Quantifying Telomere Length Using In-gel Hybridization

Frank J. Jenkins^1,2, Charles M. Kerr², Elise Fouquerel^2,3, Dana H. Bovbjerg^2,4, Patricia L. Opresko^2,3

¹Departments of Pathology and Infectious Diseases and Microbiology, University of Pittsburgh, ²University of Pittsburgh Cancer Institute, ³Department of Environmental and Occupational Health, University of Pittsburgh, ⁴Departments of Psychiatry, Psychology, Behavioral & Community Health Sciences, University of Pittsburgh

In this article, a detailed protocol for quantifying telomere length using a modified terminal restriction fragment analysis is discussed that provides fast and efficient direct measurement of telomere length. This technique can be applied to a variety of cell sources of DNA for quantifying telomere length.

Cancer Research

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Yusuf Surucu^*1, Fuat Baris Bengur^*1, Katherine S. Yang¹, Benjamin K. Schilling¹, Jocelyn S. Baker¹, Sheeba Shabbir², Renee Fisher³, Michael W. Epperly³, Joel S. Greenberger³, J. Peter Rubin¹, Asim Ejaz¹

¹Department of Plastic Surgery, University of Pittsburgh, ²Department of Forensic Medicine and Toxicology, HBS Medical and Dental College, ³Department of Radiation Oncology, University of Pittsburgh Cancer Institute

Here we present a protocol to induce radiation-induced skin fibrosis in the hind limb of mice and perform post-irradiation measurements of chronic impairment via limb excursion and gait index analyses to evaluate the functional outcome. The model elucidates radiation-related skin fibrosis mechanisms and is useful in subclinical therapeutic studies.