Our purpose is to provide researchers, consultants, decision‐makers, and other stakeholders with guidance to methods and metrics for investigating nocturnally active birds and bats in relation to utility‐scale wind‐energy development. The primary objectives of such studies are to 1) assess potential impacts on resident and migratory species, 2) quantify fatality rates on resident and migratory populations, 3) determine the causes of bird and bat fatalities, and 4) develop, assess, and implement methods for reducing risks to bird and bat populations and their habitats. We describe methods and tools and their uses, discuss limitations, assumptions, and data interpretation, present case studies and examples, and offer suggestions for improving studies on nocturnally active birds and bats in relation to wind‐energy development. We suggest best practices for research and monitoring studies using selected methods and metrics, but this is not intended as cookbook. We caution that each proposed and executed study will be different, and that decisions about which methods and metrics to use will depend upon several considerations, including study objectives, expected and realized risks to bird and bat populations, as well as budgetary and logistical considerations. Developed to complement and extend the existing National Wind Coordinating Committee document “Methods and Metrics for Assessing Impacts of Wind Energy Facilities on Wildlife” (Anderson et al. 1999), we provide information that stakeholders can use to aid in evaluating potential and actual impacts of wind power development on nocturnally active birds and bats. We hope that decision‐makers will find these guidelines helpful as they assemble information needed to support the permitting process, and that the public will use this guidance document as they participate in the permitting processes. We further hope that the wind industry will find valuable guidance from this document when 1) complying with data requirements as a part of the permitting process, 2) evaluating sites for potential development, 3) assessing impacts of operational wind‐energy facilities, and 4) mitigating local and cumulative impacts on nocturnally active birds and bats.

Stenerlöw, B., Karlsson, K. H., Cooper, B. and Rydberg, B. Measurement of Prompt DNA Double-Strand Breaks in Mammalian Cells Without Including Heat-Labile Sites. Results for Cells Deficient in Non-homologous End Joining. Radiat. Res. Ionizing radiation induces prompt single-strand breaks and double-strand breaks in DNA. In addition, labile sites are induced that can convert to breaks by heat or mild alkali. When such labile lesions are present within multiply damaged sites, additional double-strand breaks can form. Current protocols for measurement of DNA double-strand breaks involve a lysis step at elevated temperature, and consequently breaks from heat-labile sites will be generated during lysis and will be included in the measurement. However, such sites may not develop into breaks within the cell and may not need DNA double-strand break repair processes for elimination. We present here a new lysis and pulsed-field gel electrophoresis protocol that is carried out entirely at 0-4°C and thus avoids inclusion of heat-labile sites into the measurement. The new recommended lysis procedure involves two steps: the first step includes proteinase-K, which has sufficient activity at 0°C to support lysis, and the second step includes a high salt buffer to strip the DNA from histones and other proteins. By various tests we conclude that lysis is sufficient with this procedure to allow accurate determination of double-strand breaks by pulsed-field gel electrophoresis. Using the new protocol, it was found that heat-labile sites account for 30% of the initial number of double-strand breaks measured by conventional protocols after low LET radiation. In addition we show that heat-labile sites that can convert to double-strand breaks are repaired with fast kinetics and are nearly completely eliminated after 1 hr at 37°C. A study of cells deficient in non-homologous end joining reveals that the residual fast repair response typically seen in such cells is solely due to repair at heat-labile sites and is not due to repair of prompt DSBs. 2

There is an emerging body of evidence implicating iron in carcinogenesis and in particular colorectal cancer, but whether this involves Wnt signalling, a major oncogenic signalling pathway has not been studied. We aimed to determine the effect of iron loading on Wnt signalling using mutant APC (Caco-2 and SW480) and wild-type APC (HEK-293 and human primary fibroblasts) containing cell lines. Elevating cellular iron levels in Caco-2 and SW480 cells caused increased Wnt signalling as indicated by increased TOPFLASH reporter activity, increased mRNA expression of two known targets, c-myc and Nkd1, and increased cellular proliferation. In contrast wild-type APC and beta-catenin-containing lines, HEK 293 and human primary fibroblasts were not responsive to iron loading. This was verified in SW480 cells that no longer induced iron-mediated Wnt signalling when transfected with wild-type APC. The cell line LS174T, wild type for APC but mutant for beta-catenin, was also responsive suggesting that the role of iron is to regulate beta-catenin. Furthermore, we show that E-cadherin status has no influence on iron-mediated Wnt signalling. We thus speculate that excess iron could exacerbate tumorigenesis in the background of APC loss, a common finding in cancers.

Characteristics of nocturnal bird migration are poorly understood for many regions of the United States. This information will be critical in areas where wind power projects are proposed. We used portable marine radar to conduct a nocturnal bird migration study at multiple sites along the Allegheny Front, West Virginia, on 45 nights during autumn 2003, to document migration characteristics at a proposed wind power project. Nocturnal passage rates were highly variable among nights, ranging from 8 to 852 targets/km/hour, with a seasonal mean of 241 ± 33 targets/km/hour at the primary (central) study site and 199 targets/km/hour for the entire proposed development. Mean flight altitudes also were highly variable among nights, ranging from 214 to 769 m above ground level (agl), with a mean flight altitude of 410 ± 2 m agl. Flight directions indicated that most migrants crossed, rather than followed, the Allegheny Front ridgeline. We believe portable marine radars, when coupled with a rigorous study design, can collect important baseline information on avian migration and address site specific questions posed at proposed developments. Concurrent collection of low‐altitude migration and avian fatality data could help elucidate which metrics are most useful for predicting avian fatalities at wind power developments.