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University of Pittsburgh Cancer Institute

3 ARTICLES PUBLISHED IN JoVE

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Biology

Quantitative, Real-time Analysis of Base Excision Repair Activity in Cell Lysates Utilizing Lesion-specific Molecular Beacons
David Svilar 1,2, Conchita Vens 3, 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.

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Cancer Research

Modified Terminal Restriction Fragment Analysis for Quantifying Telomere Length Using In-gel Hybridization
Frank J. Jenkins 1,2, Charles M. Kerr 2, 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.

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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. Yang 1, Benjamin K. Schilling 1, Jocelyn S. Baker 1, Sheeba Shabbir 2, Renee Fisher 3, Michael W. Epperly 3, Joel S. Greenberger 3, J. Peter Rubin 1, Asim Ejaz 1
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.

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