Convection-permitting Weather Research and Forecasting (WRF) Model forecasts with 3-km horizontal grid spacing were produced for a 50-member ensemble over a domain spanning three-quarters of the contiguous United States between 25 May and 25 June 2012. Initial conditions for the 3-km forecasts were provided by a continuously cycling ensemble Kalman filter (EnKF) analysis–forecast system with 15-km horizontal grid length. The 3-km forecasts were evaluated using both probabilistic and deterministic techniques with a focus on hourly precipitation. All 3-km ensemble members overpredicted rainfall and there was insufficient forecast precipitation spread. However, the ensemble demonstrated skill at discriminating between both light and heavy rainfall events, as measured by the area under the relative operating characteristic curve. Subensembles composed of 20–30 members usually demonstrated comparable resolution, reliability, and skill as the full 50-member ensemble. On average, deterministic forecasts initialized from mean EnKF analyses were at least as or more skillful than forecasts initialized from individual ensemble members “closest” to the mean EnKF analyses, and “patched together” forecasts composed of members closest to the ensemble mean during each forecast interval were skillful but came with caveats. The collective results underscore the need to improve convection-permitting ensemble spread and have important implications for optimizing EnKF-initialized forecasts.

The Advanced Research version of the Weather Research and Forecasting Model (ARW) coupled with an urban canopy model is used to investigate the potential of vegetative (green) roof technology to mitigate against ongoing climate warming and continued urban sprawl for a day representing average summer conditions in late-twenty-first-century Chicago, Illinois. Effects related particularly to human health hazards resulting from excessive heat and high pollution concentrations are emphasized. Continued expansion of the urban environment over the next century is shown to lead to an expansion of the warming signal across the metropolitan region. Widespread adoption of vegetative rooftops, through increased albedo and evapotranspiration, reduces temperatures in the urban environment by as much as 38C, an effect similar to the simpler but less appealing alternative of employing painted or other reflective rooftop structures (e.g., white roofs). A significant limitation to the green roof approach for the case studied is that the increase in moisture resulting from transpiration leads to only marginal cooling when apparent temperatures are considered. An additional complication arises in that the reduced temperatures alter the lake-breeze circulation, potentially reducing circulation of pollutants into the city core, but also reducing natural cooling in the most urbanized areas during the climatologically warmest hours. Future work that evaluates these impacts over a broader range of synoptic settings, documents changes in the planetary boundary layer structure and attendant pollution, and considers the multiple-day dependence of these effects is needed.

Thymidine Kinase 1 (TK1) is primarily known as a cancer biomarker with good prognostic capabilities for both hematological and solid malignancies. However, recent studies targeting TK1 at protein and mRNA levels have shown that TK1 may be useful as a therapeutic target. In order to examine the use of TK1 as a therapeutic target, it is necessary to develop therapeutics specific for it. Single domain antibodies (sdAbs), represent an exciting approach for the development of immunotherapeutics due to their cost-effective production and higher tumor penetration than conventional antibodies. In this study, we isolated sdAb fragments specific to human TK1 from a human sdAb library. A total of 400 sdAbs were screened through 5 rounds of selection by monoclonal phage ELISA. The most sensitive sdAb fragments were selected as candidates for preclinical testing. The sdAb fragments showed specificity for human TK1 in phage ELISA, Western blot analysis and had an estimated limit of detection of 3.9 ng/ml for the antibody fragments 4-H-TK1_A1 and 4-H-TK1_D1. The antibody fragments were successfully expressed and used for detection of membrane associated TK1 (mTK1) through flow cytometry on cancer cells [lung (~95%), colon (~87%), breast (~53%)] and healthy human mononuclear cells (MNC). The most sensitive antibody fragments, 4-H-TK1_A1 and 4-H-TK1_D1 were fused to an engineered IgG1 Fc fragment. When added to cancer cells expressing mTK1 co-cultured with human MNCs, the anti-TK1-sdAb-IgG1_A1 and D1 were able to elicit a significant antibody-dependent cell-mediated cytotoxicity (ADCC) response against lung cancer cells compared to isotype controls (P<0.0267 and P<0.0265, respectively). To our knowledge this is the first time that the isolation and evaluation of human anti-TK1 single domain antibodies using phage display technology has been reported. The antibody fragments isolated here may represent a valuable resource for the detection and the targeting of TK1 on tumor cells.

This study explores the potential therapeutic use of monoclonal antibodies against the tumor proliferation biomarker TK1 for the immunotargeting of lung and breast cancer cells. Recent clinical trials have led to the approval of monoclonal antibodies for the treatment of several solid tumors including lung and breast tumors. However, a common limitation that antibody-based therapies face is that a significant portion of the current tumor targets are expressed on normal tissues, thus creating off-target tumor effects. TK1 is a tumor proliferation biomarker that is up-regulated in malignant tissues. Recently, we have reported the expression of TK1 on the cell membrane of several cancer cell lines, mononuclear cells (MNC) from patients with leukemia and cells from breast and colon tumors. No membrane expression of TK1 was found on normal cells. To further explore the potential of TK1 as an immunotherapeutic target, we evaluated the capacity of a novel panel of anti TK1 antibodies to target TK1 on the cell membrane of lung and breast cancer cells and to elicit an antibody-dependent cell-mediated-cytotoxicity (ADCC) response in vitro. Antibodies previously developed in our lab were validated to specifically target six different regions of the TK1 molecule. Four different antibodies targeting three different regions in the TK1 molecule were selected for ADCC experiments. Nuclear restricted GFP versions of the NCI-H460 and MDA-MB-231 cell lines were utilized in conjunction with the real-time cell imaging system ImageXpress® Pico to measure the ADCC response. Mono nuclear cells (MNCs) from healthy donors were co-cultured with target cells and 10ug/ml of each anti-TK1 antibody were added to the media. An antibody with the same isotype than the TK1 antibodies, but that doesn't bind to any human surface antigen was utilized as a control. All ADCC responses elicited by the anti-TK1 antibodies were observed between 24-72hrs. The highest ADCC responses were elicited by antibodies 4G10 and 1B12 in both breast and lung cancer cell lines while a minimal response was observed with antibodies 7D1 and 3B2E11. A 50-60% increment in the ADCC response against NCI-H460 cells was observed with antibody 4G10 in comparison with the isotype control (p= 0.001). A 40% increase in the ADCC response against MDA-MB-231 cells was observed using antibody 1B12 (p = 0.003) compared to the its isotype control. This preliminary data suggests that anti TK1 monoclonal antibodies can be used for the immunotargeting of tumor cells expressing membrane associated TK1. The further exploration of TK1 as a tumor target could lead to the development of new TK1-based immunotherapies. Citation Format: Edwin J. Velazquez, Jordan D. Cress, Kathryn R. Smith, Taylor D. Brindley, Gajendra Shrestha, Richard A. Robison, Kim L. O'Neill. Monoclonal antibodies against thyimidine kinase 1 for the immunotargeting of lung and breast cancer cells, a preclinical evaluation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 543.

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