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The evolution from microarrays to transcriptome deep-sequencing (RNA-seq) and from RNA interference to gene knockouts using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) and Transcription Activator-Like Effector Nucleases (TALENs) has provided a new experimental partnership for identifying and quantifying the effects of gene changes on drug resistance. Here we describe the results from deep-sequencing of RNA derived from two cytarabine (Ara-C) resistance acute myeloid leukemia (AML) cell lines, and present CRISPR and TALEN based methods for accomplishing complete gene knockout (KO) in AML cells. We found protein modifying loss-of-function mutations in Dck in both Ara-C resistant cell lines. CRISPR and TALEN-based KO of Dck dramatically increased the IC50 of Ara-C and introduction of a DCK overexpression vector into Dck KO clones resulted in a significant increase in Ara-C sensitivity. This effort demonstrates the power of using transcriptome analysis and CRISPR/TALEN-based KOs to identify and verify genes associated with drug resistance.

India has ambitious goals for high utilization of variable renewable power from wind and solar, and deployment has been proceeding at a rapid pace. The western state of Gujarat currently has the largest amount of solar generation of any Indian state, with over 855 Megawatts direct current (MWDC). Combined with over 3,240 MW of wind, variable generation renewables comprise nearly 18% of the electric-generating capacity in the state. A new historic 10-kilometer (km) gridded solar radiation data set capturing hourly insolation values for 2002-2011 is available for India. We apply an established method for downscaling hourly irradiance data to one-minute irradiance data at potential PV power production locations for one year, 2006. The objective of this report is to characterize the intra-hour variability of existing and planned photovoltaic solar power generation in the state of Gujarat (a total of 1.9 gigawatts direct current (GWDC)), and of five possible expansion scenarios of solar generation that reflect a range of geographic diversity (each scenario totals 500-1,000 MW of additional solar capacity). The report statistically analyzes one year's worth of power variability data, applied to both the baseline and expansion scenarios, to evaluate diurnal and seasonal power fluctuations, different timescales of variability (e.g.,more » from one to 15 minutes), the magnitude of variability (both total megawatts and relative to installed solar capacity), and the extent to which the variability can be anticipated in advance. The paper also examines how Gujarat Energy Transmission Corporation (GETCO) and the Gujarat State Load Dispatch Centre (SLDC) could make use of the solar variability profiles in grid operations and planning.« less

This report summarizes the results of an assessment and analysis of renewable energy opportunities conducted for the U.S. Department of the Interior, Bureau of Reclamation by the National Renewable Energy Laboratory. Tasks included assessing the suitability for wind and solar on both a utility and facility scale.

This paper assesses alterations in collagen metabolism following thrombolytic therapy of acute myocardial infarction with tissue-plasminogen activator. Sequential serum measurements of the amino-terminal propeptide of type III procollagen (S-PIIINP) and the carboxyterminal propeptide of type I collagen (S-PICP) in patients suspected of acute myocardial infarction randomized to tissue-plasminogen activator or placebo were used. S-PIIINP increased at 3h in patients with acute myocardial infarction treated with tissue-plasminogen activator (P < 0.05). S-PIIINP was higher in patients treated with tissue-plasminogen activator compared with placebo-treated patients at 3 and 6 h (P < 0.05). S-PICP decreased independently of therapy and diagnosis. Tissue-plasminogen activator, therefore, induces breakdown of collagen, some of which is located in the wall of atheromatous arteries. Vascular patency following thrombolytic therapy may partly be mediated by breakdown of thrombogenic collagen in the vessel wall. The findings may suggest a role for S-PIIINP as a non-invasive indicator of the risk of reocclusion.

This guide presents an overview of the process for successfully planning for and installing solar technology on a federal site. It is specifically targeted to managers of federal buildings and sites, contracting officers, energy and sustainability officers, and regional procurement managers. The solar project process is outlined in a concise, easy-to-understand, step-by-step format. Information includes a brief overview of legislation and executive orders related to renewable energy and the compelling reasons for implementing a solar project on a federal site. It also includes how to assess a facility to identify the best solar installation site, project recommendations and considerations to help avoid unforeseen issues, and guidance on financing and contracting options. Case studies with descriptions of successful solar deployments across multiple agencies are presented. In addition, detailed information and sample documents for specific tasks are referenced with Web links or included in the appendixes. The guide concentrates on distributed solar generation and not large, centralized solar energy generation.

To evaluate MRI for assessment of inflammation, destruction and prognosis in rheumatoid arthritis (RA), 26 RA patients, randomized to disease-modifying anti-rheumatic drug (DMARD) therapy alone or in combination with oral prednisolone, were followed for one year with contrast-enhanced MRI of the dominant wrist (months zero, three, six and 12), conventional radiography and clinical and biochemical examinations. Significant synovial membrane volume reductions were observed in both groups, earliest in the DMARD + prednisolone group. The rate of erosive progression on MRI was highly correlated with baseline and area-under-curve (AUC)-values of synovial membrane volume, but not with baseline or AUC-values of local or global clinical or biochemical parameters, nor with +/- prednisolone. MRI was more sensitive than radiography as regards detection of progressive bone destruction (22 versus 12 new bone erosions). MRI may prove valuable as marker of joint disease activity and destruction and, perhaps, prognosis in RA.

Abstract 2489 One frequently reported change to acute myeloid leukemia (AML) cells, as they become resistant to arabinoside cytarabine (Ara-C), is the downregulation of deoxycytidine kinase (Dck) (Veuger et al, Blood, 2000, Shi et al, Pharmacogenetics, 2004, Cai et al, Cancer Res, 2008, Song et al, Int J Oncol, 2009, Rathe et al, Leukemia, 2010). Dck is the rate limiting enzyme in the phosphorylation and activation of Ara-C. Transcriptome deep sequencing (RNA-seq) was performed on the transcriptome of two Ara-C resistant murine leukemia cell lines (B117H and B140H), as well their Ara-C sensitive parental cell lines (B117P and B140P). Previously published microarray analysis identified the downregulation of Dck as the primary common change in the B117H and B140H cell lines, when compared to their parental cell lines (Rathe et al, Leukemia, 2010). Analysis of the RNA-seq results identified an alternatively spliced form of Dck in the B117H cell line secondary to a genomic deletion/rearrangement, as well as a loss of heterozygosity. Indeed, knockdowns of Dck in the parental cell lines demonstrated that downregulation of Dck can result in a significant increase in the IC50 of Ara-C, consistent with the increase in Ara-C resistance displayed by the B117H and B140H cells. The RNA-seq analysis showed a number of significant expression changes that did not appear on the microarray analysis. These changes include the significant upregulation of specific isoforms of deoxyguanosine kinase (Dguok) and protein phosphatase 2 (formerly 2A), regulatory subunit A (PR65), beta isoform (Ppp2r1b). It has already been shown that downregulation of Dguok results in the upregulation of Dck (Franco et al, Exp Cell Res, 2007), thereby demonstrating an important balance between these two molecules. Ppp2r1b is a putative deactivator of Dck (Smal et al, Biochem Pharmacol, 2004), which may assist in the prevention of Ara-C metabolic activation and thereby contribute to resistance. This effort demonstrates the power of using RNA-seq transcriptome analysis to identify drug targets for the treatment of AML and suggests multiple genetic/epigenetic mechanisms contribute to alterations in Dck activity during the acquisition of Ara-C resistance. Disclosures: Largaespada: NeoClone Biotechnology, Inc.: Co-founder, Consultancy, Equity Ownership; Discovery Genomics Inc.: Co-founder, Consultancy, Equity Ownership.