This report provides summarized physical, chemical, and biological data collected during a study of the effects of urbanization on stream ecosystems as part of the U.S. Geological Survey’s National Water-Quality Assessment study. The purpose of this study was to examine differences in biological, chemical, and physical characteristics of streams across a gradient of urban intensity. Thirty sites were selected along an urbanization gradient that represents conditions in the North Carolina Piedmont ecoregion, including the cities of Raleigh, Durham, Cary, Greensboro, Winston-Salem, High Point, Asheboro, and Oxford. Data collected included streamflow variability, stream temperature, instream chemistry, instream aquatic habitat, and collections of the algal, macroinvertebrate, and fish communities. In addition, ancillary data describing land use, socioeconomic conditions, and urban infrastructure were compiled for each basin using a geographic information system analysis. All data were processed and summarized for analytical use and are presented in downloadable data tables, along with the methods of data collection and processing.