text: REPORTS minor contribution to polymerization from the monomeric state. In contrast, the arms are of minor overall importance to binding but contribute to polymerization of monomeric actin when covalently attached to the globular domain. These deletion results appear to segregate SipA function directly along structural lines. The ability of SipA to polymerize monomeric actin is dependent on the extended arms, which bind spatially distant actin molecules, whereas the globular core functions primarily to bind to preexisting filaments, probably because the core is unable to effectively tether molecules in opposing strands. In combination, the core and tethering arms enable SipA to both induce and stabilize the filamentous form of actin. This division of function provides an explanation for the ability of certain "one-armed" constructs to retain some ability to polymerize G-actin, because the core binding with an additional arm tethering may confer enough additional stability to the filament to enhance polymerization. The presence of the "arm" nonglobular density may be expected to create a molecule with an elongated appearance in solution, as is observed by EM (10) in actin complexes with SipA446 – 684 and by small-angle x-ray scattering (13). We determined the hydrodynamic radius of various SipA constructs (Fig. 3F) and found not only that the ordered core (as found in the crystals) was globular in solution but also that SipA constructs with additional polypeptide arms were elongated in solution and became increasingly elongated as more arm polypeptide was added (Fig. 3F). As estimated by the frictional coefficient, the larger SipA molecules were as elongated as fibronectin and tenascin (14). To further probe this hypothesis of flexible polypeptides at the N- and C-termini of the molecule, we subjected a larger construct, SipA425– 684, to limited proteolysis with the relatively nonspecific protease subtilisin (Fig. 3E) (11). The protease cut very effectively until it approached a construct that was ordered in the crystals (SipA513– 657), producing trimmed constructs that when subcloned and overexpressed were highly soluble, stable, and active to bind F-actin (Fig. 3, E and F). These observations are consistent with a model that proposes a well-folded, protease-resistant globular domain with protease-sensitive polypeptides extending from its termini. Thus, on the basis of our EM, proteolytic digestion, and hydrodynamic radius data, we show that the arm polypeptides that are critical to activity contribute to an elongated appearance in actin-SipA complexes, are associated with proteolytically accessible peptide, and elongate SipA in solution. Because eukaryotic proteins like nebulin are also elongated in tethering actin subunits in the filament, the lack of sequence or structural similarity between SipA and such host molecules indicates that it probably represents another case of convergent evolution in bacterial pathogenesis (10, 15). 1. T. Pang, Z. A. Bhutta, B. B. Finlay, M. Altwegg, Trends Microbiol. 3, 253 (1995). 2. T. J. Torok et al., JAMA 278, 389 (1997). 3. J. E. Galan, A. Collmer, Science 284, 1322 (1999). 4. G. R. Cornelis, F. Van Gijsegem, Annu. Rev. Microbiol. 54, 735 (2000). 5. J. E. Galan, D. Zhou, Proc. Natl. Acad. Sci. U.S.A. 97, 8754 (2000). 6. R. M. Tsolis et al., Infect. Immun. 68, 3158 (2000). 7. S. Zhang et al., Infect. Immun. 70, 3843 (2002). 8. D. Zhou, M. S. Mooseker, J. E. Galan, Science 283, 2092 (1999). 9. E. J. McGhie, R. D. Hayward, V. Koronakis, EMBO J. 20, 2131 (2001). 10. V. E. Galkin et al., Nature Struct. Biol. 9, 518 (2002). 11. Materials and methods are available as supporting material on Science Online. 12. L. Holm, C. Sander, J. Mol. Biol. 233, 123 (1993). 13. K. Mitra, D. Zhou, J. E. Galan, FEBS Lett. 482, 81 (2000). 14. G. Schurmann, J. Haspel, M. Grumet, H. P. Erickson, Mol. Biol. Cell 12, 1765 (2001). 15. N. Lukoyanova et al., Curr. Biol. 12, 383 (2002). 16. Figures were prepared with Molscript (17), Raster3D (18), Swiss-PdbViewer (19), and Ribbons (20). 17. P. J. Kraulis, Appl. Crystallogr. 24, 946 (1991). 18. E. A. Merritt, D. J. Bacon, Methods Enzymol. 277, 505 (1997). 19. N. Guex, M. C. Peitsch, Electrophoresis 18, 2714 (December 1997). 20. M. M. Carson, Methods Enzymol. 277, 493 (1997). 21. This work was funded by Public Health Services grants (to C.E.S. and E.H.E.) and through a BurroughsWellcome Investigators in Pathogenesis of Infectious Disease award to C.E.S. Coordinates have been deposited to the Protein Data Bank under ID code1Q5Z. Supporting Online Material www.sciencemag.org/cgi/content/full/301/5641/1918/ DC1 Materials and Methods Fig. S1 Table S1 References 25 June 2003; accepted 14 August 2003 References and Notes R2D2, a Bridge Between the Initiation and Effector Steps of the Drosophila RNAi Pathway Qinghua Liu,1 Tim A. Rand,1 Savitha Kalidas,2 Fenghe Du,1 Hyun-Eui Kim,1 Dean P. Smith,2 Xiaodong Wang1* The RNA interference (RNAi) pathway is initiated by processing long doublestranded RNA into small interfering RNA (siRNA). The siRNA-generating enzyme was purified from Drosophila S2 cells and consists of two stoichiometric subunits: Dicer-2 (DCR-2) and a previously unknown protein that we named R2D2. R2D2 is homologous to the Caenorhabditis elegans RNAi protein RDE-4. Association with R2D2 does not affect the enzymatic activity of DCR-2. Rather, the DCR-2/R2D2 complex, but not DCR-2 alone, binds to siRNA and enhances sequence-specific messenger RNA degradation mediated by the RNA-initiated silencing complex (RISC). These results indicate that R2D2 bridges the initiation and effector steps of the Drosophila RNAi pathway by facilitating siRNA passage from Dicer to RISC. RNA interference (RNAi) is a form of posttranscriptional gene silencing whereby double-stranded RNA (dsRNA) molecules trigger the sequence-specific degradation of cognate mRNA (1–3). The biological importance of RNAi is underscored by its wide conservation throughout metazoans and the existence of closely related systems in plants (called cosuppression) and fungi (called quelling) (4). Emerging evidence indicates that the RNAi and related pathways function 1 Howard Hughes Medical Institute and Department of Biochemistry, 2Department of Pharmacology and Center for Basic Neuroscience, University of Texas (UT) Southwestern Medical Center at Dallas, Dallas, TX 75390, USA. *To whom correspondence should be addressed. Email: firstname.lastname@example.com in many fundamental biological processes, including antiviral defense, development, and maintenance of genomic stability (4). The Drosophila RNAi pathway consists of initiation and effector steps. First, long dsRNA molecules are cleaved into 21- to 23-nucleotide (nt) small interfering RNA (siRNA) duplexes (5– 8). Secondly, the siRNA is incorporated into a nuclease complex named RNA-initiated silencing complex (RISC) and functions as a guide RNA to direct RISC-mediated sequence-specific mRNA degradation (6, 9 –11). The endonucleases that process dsRNA have been identified as Dicers, a family of large noncanonical ribonuclease (RNase) III enzymes (5). Although only one Dicer enzyme is found in Caenorhabditis elegans and humans, two, DCR-1 and DCR-2, have www.sciencemag.org SCIENCE VOL 301 26 SEPTEMBER 2003 1921 REPORTS Fig. 1. (A) Purification of the siRNA-generating activity. Individual fractions from the final gel filtration column were assayed for Dicer activity (12) (top) or resolved by SDS–polyacrylamide gel electrophoresis (PAGE) followed by silver staining (bottom). Arrows indicate the positions of dsRNA and siRNA (top) or DCR-2 and R2D2 proteins (bottom). (B) A protein sequence alignment of R2D2 (D. melanogaster) and RDE-4 (C. elegans). The two consensus dsRNA-binding motifs are boxed. Identical residues are shaded in black and conserved residues in gray. (C) Immunodepletions were performed in S100 (lane 1 and 5) with the use of the preimmune serum (lane 2 and 6) or anti-R2D2 serum (lane 3 and 7). DCR-1, DCR-2, or R2D2 proteins were detected in the supernatants by Western blotting with the use of the corresponding antibody (left). The Dicer assays were carried out with 1 l of each supernatant (right). A 21-nt radiolabeled siRNA marker (M) is shown in lane 4. (D) S100 was fractionated by a Q-Sepharose column. DCR-1, DCR-2, and R2D2 proteins were detected by Western blotting (top), whereas the siRNA-generating activities were measured among different fractions (bottom). Fig. 2. (A) GFP transgenic S2 cells were soaked in media containing luciferase (LUC), DCR-1, or DCR-2 dsRNA for 72 hours, and lysates were prepared. The levels of DCR-1, DCR-2, R2D2, and GFP proteins were measured by Western blotting. DCR-1 and DCR-2 Western used the same blot, and so did R2D2 and GFP Western. (B) Total RNA was isolated from S2(GFP) cells treated with GFP (lanes 1 and 3) or DCR-2 (lanes 2 and 4) dsRNA. Northern blots (same membrane) were performed to detect DCR-2 (lanes 1 and 2) or R2D2 (lanes 3 and 4) mRNA. The asterisk represents degradation products of DCR2 mRNA. (C) The siRNA-generating activities were compared among lysates prepared from GFP (lanes 1 and 2), DCR-1 (lanes 3 and 4), or DCR-2 (lanes 5 and 6) dsRNA-treated S2 cells. (D) Lysates were prepared from wild-type flies (lane 1), the starting P element strain (lane 2), and the r2d2 mutant flies derived by P element mobilization (lane 3). Western blots were performed with the use of antibody against R2D2 and antibody against -tubulin as loading control. (E) Wild-type or r2d2 mutant embryos expressing GFP were injected with GFP dsRNA. Images shown include a wild-type embryo injected with buffer (lane 1, –) or GFP dsRNA (lane 2, ) or an r2d2 mutant embryo injected with buffer (lane 3, –) or GFP dsRNA (lane 4, ). In total, 46 of 52 wild-type embryos and 0 of 30 r2d2 mutant embryos showed silencing of GFP expression. been identified in Drosophila (5). It remains unclear how siRNA is transferred from Dicer to RISC. We studied this problem by purifying the siRNA-generating activity from the cytoplasmic (S100) extract of S2 cells through a six-step chromatographic procedure (12). A single major peak of activity was observed at all steps and was followed throughout purification. Two proteins, 190 kD and 36 kD, showed perfect correlation with the enzymatic activity after the final gel filtration step (Fig. 1A). They were identified by mass spectrometry to be DCR-2 and a previously unknown protein (Flybase CG7138), respectively. We named this protein R2D2 because it contains two dsRNA-binding domains (R2) and is associated with DCR-2 (D2). R2D2 bears 20.9% identity and 33.4% similarity to the C. elegans RNAi protein RDE-4, which also contains tandem dsRNA-binding domains and interacts with Dicer (Fig. 1B) (13, 14 ). To confirm our purification results, we performed immunodepletion experiments with antiserum directed against the carboxylterminal 150 amino acids of R2D2. This R2D2 antibody depleted both R2D2 and DCR-2 proteins and removed the majority of siRNA-generating activity from S100, whereas the level of DCR-1 remained unchanged in the supernatant (Fig. 1C). Furthermore, when S100 was fractionated on a QSepharose column (Amersham), DCR-2 and 1922 26 SEPTEMBER 2003 VOL 301 SCIENCE www.sciencemag.org REPORTS Fig. 3. (A) A Coomassie-stained SDS-PAGE gel showing purified recombinant DCR-2 (lane 1), wild-type DCR-2/R2D2 (lane 2), or mutant DCR-2/R2D2M complex proteins. (B) ( Top) A schematic diagram of the domain structure of R2D2 protein. The shaded boxes refer to dsRNA-binding (dsrb) domains. (Middle) The consensus dsrb motif sequence is shown. Two conserved alanines (A) were substituted by lysines (K) in both dsrb motifs in the mutant R2D2 protein. G, glycine; S, serine; L, leucine; and x, any amino acid. (Bottom) A dsRNA-binding assay by using poly (inosine):poly (cytidine)-conjugated beads to precipitate in vitro–translated 35S-labeled wild-type (WT, lanes 1 and 2) and mutant (M, lanes 3 and 4) R2D2 protein (12). Twenty-five percent of input (I) and all of bound (B) proteins were resolved by SDS-PAGE, detected and quantified by phosphorimaging. The percentage of bound/input protein is indicated below the lanes. (C) DCR-2 is ATP-dependent and shows equivalent siRNA-generating activity as DCR2/R2D2. The concentration of recombinant proteins was measured by Bradford assay and indicated above the lanes. Dicer assays were performed in the absence (lanes 1 to 5) or presence (lanes 6 to 10) of ATP. Arrows indicate dsRNA and arrowheads, siRNA. (D) DCR-2/R2D2M is as active as the wild-type complex. A series of dilutions of wild-type DCR-2/R2D2 (lanes 1 to 5) and mutant DCR-2/R2D2M (lanes 6 to 10) complex proteins were compared for Dicer activity in the presence of ATP. Fig. 4. (A) A gel shift assay showing a siRNA-binding activity in S100 (12). Radiolabeled siRNA was incubated with buffer alone (lane 1) or 40 g of S100 (lane 2) in the absence of ATP. (B) Gel shift assays were performed in the presence of antibodies against DCR-2 (lane 2) or R2D2 (lane 4), or their preimmune sera (lanes 1 and 3). Short and long arrows indicate the mobility shift and supershift, respectively. The free siRNA ran off the gel. (C) Gel shift assays were performed with 5 g of S100 (lane 1) and 250 ng of purified DCR-2 (lane 2), DCR-2/R2D2 (lane 3), or DCR-2/R2D2M (lane 4) recombinant proteins in the absence of ATP. (D) Gel shift assays were performed as in (C) but with the presence of ATP to decrease nonspecific crosslinking after UV exposure (12), resolved by SDS-PAGE, transferred to a cellulose membrane, and exposed to x-ray film (lanes 4 to 6). The same membrane was Western blotted with the antibody against His (lanes 1 to 3). Arrows indicate the positions of DCR-2 and R2D2 (lanes 1 to 3), or siRNA-crosslinked DCR-2 and R2D2 (lanes 4 to 6). The siRNA1 and siRNA2 refer to one or two strands of siRNA, respectively. (E) Wild-type DCR-2/R2D2 complex was used for the gel shift assay and UV crosslinking. Half of the reaction was treated with RNase A/T1, and half was untreated. Arrows and arrowheads refer to DCR-2 and R2D2 proteins that are crosslinked to full-length or RNase-digested siRNA, respectively. R2D2, but not DCR-1, correlated perfectly with the siRNA-generating activity (Fig. 1D). To determine the contribution of DCR-1 and DCR-2 to siRNA production in Drosophila cells, we depleted either protein from S2 cells by dsRNA soaking (Fig. 2A). Interestingly, DCR-2 dsRNA also caused a substantial reduction in the level of R2D2 protein. This was not simply due to cross-targeting because the level of R2D2 mRNA was unaffected (Fig. 2B). Likewise, R2D2 dsRNA also reduced, although not as dramatically, the level of DCR-2 protein (15). These results indicate that DCR-2 and R2D2 form a stable complex and that either protein alone is unstable. Although depletion of DCR-1 made no difference, knocking down DCR-2 reduced the siRNA-generating activity by fivefold in lysates and interfered with RNAi in vivo (Fig. 2C and fig. S1). Together, these results demonstrate that the DCR-2/ R2D2 complex is the principal siRNAgenerating enzyme responsible for initiation of RNAi in Drosophila S2 cells. www.sciencemag.org SCIENCE VOL 301 26 SEPTEMBER 2003 1923 REPORTS To determine whether R2D2 is required for RNAi in vivo, we generated r2d2 deletion mutant flies by P element mobilization (Fig. 2D) and crossed them with transgenic flies expressing green fluorescent protein (GFP) under the ubiquitin promoter to derive homozygous r2d2; Ub-GFP mutant flies (16). We then collected 0- to 2-hour wild-type or r2d2 mutant embryos for microinjection of GFP dsRNA. Whereas introduction of GFP dsRNA effectively silenced GFP expression in wild-type embryos, r2d2 mutant embryos were completely defective for the dsRNAinitiated RNAi response (Fig. 2E). The siRNA-generating activity was reconstituted in vitro with the use of purified His6tagged DCR-2 and R2D2 recombinant proteins expressed in insect cells (Fig. 3A). A mutant form of R2D2 was also created by point mutations in the dsRNA-binding domains that abolish its ability to bind dsRNA (Fig. 3B). As shown in Fig. 3C, DCR-2 protein alone efficiently cleaved dsRNA into siRNA in an adenosine triphosphate (ATP)– and dose-dependent manner. The DCR-2/R2D2 complex was also ATP-dependent and showed activity equivalent to DCR-2 alone. We also performed kinetic studies with DCR-2 and DCR-2/R2D2 recombinant proteins and found no statistically significant difference in their Km (a measure of substrate affinity) or Kcat (a measure of catalytic activity) (15). Furthermore, the mutant DCR-2/ R2D2M complex was as active in siRNA production as the wild-type complex (Fig. 3D). These results indicate that association with R2D2 does not affect the ability of DCR-2 to recruit or cleave dsRNA. This finding is inconsistent with the proposed function of RDE-4 to recruit dsRNA to DCR-1 (the single Dicer in worms) for processing (13). However, R2D2 may stabilize DCR-2 and thereby positively regulate siRNA production in Drosophila cells. Consistently, DCR-2 and R2D2 were expressed at much higher levels in insect cells when expressed jointly than separately (15). To follow the fate of siRNA, we developed a gel shift assay to identify proteins that interact with siRNA. When radiolabeled synthetic siRNA was incubated with S2 lysate, a distinct mobility shift was observed on a native polyacrylamide gel (Fig. 4A). The formation of this siRNA-protein complex did not require ATP hydrolysis because it could be carried out efficiently at 4°C (15). Furthermore, when S100 was fractionated by a QSepharose column, the peak of siRNAbinding activity correlated well with that of the siRNA-generating activity (15). We then performed the gel shift assays in the presence of antibodies against DCR-2 or R2D2. Both antibodies resulted in a supershift that was absent when their preimmune sera were used instead (Fig. 4B). This indicated that DCR-2 and R2D2 were present in this siRNA-protein complex. Purified DCR-2, DCR-2/R2D2, and DCR-2/R2D2M recombinant proteins were examined for siRNA binding by the gel shift assay. As shown in Fig. 4C, wild-type DCR-2/R2D2 complex, but not DCR-2 alone, bound to siRNA and produced a mobility shift indistinguishable from that of S100. The ability of the DCR-2/R2D2 complex to bind siRNA was greatly diminished by point mutations within the two dsRNAbinding domains of R2D2 (Fig. 4C, lanes 3 and 4). Thus, R2D2 is important for binding to the product (siRNA) rather than the substrate (dsRNA) of DCR-2. Furthermore, when DCR-2/R2D2 proteins and radiolabeled siRNA were exposed to ultraviolet (UV) light, both DCR-2 and R2D2 were crosslinked to siRNA (Fig. 4D, lanes 4 Fig. 5. (A) Partial purification of RISC. The Dicer assays (top) and the dsRNA- (middle) and siRNAinitiated (bottom) RISC assays were performed with the use of the supernatant (S) or pellet resuspension (P) of 5% (lanes 1 and 2), 10% (lanes 3 and 4), and 15% (lanes 5 and 6) PEG precipitation. The initiation time was 45 min at 30°C. For siRNA-initiated RISC assays, a 200-nt mRNA substrate containing a single let-7 target site was radiolabeled at the 5 cap and used as previously described (17). The 5 cleavage product is 115 nt. For dsRNA-initiated RISC assays, a radiolabeled mRNA containing tandem let-7 target sites was used to ensure maximal RISC activity. The 5 cleavage products range from 100 to 150 nt. The 130-bp dsRNA (3 nM) used to trigger RISC activity contained four direct repeats of the let-7 sequence. (B) The dsRNA-initiated RISC assays were performed in 10% PEG supernatant alone (lane 1) or in combination with 3 nM or 10 nM of recombinant DCR-2 (lanes 2 and 3), DCR-2/ R2D2 (lanes 4 and 5), or DCR-2/ R2D2M (lanes 6 and 7) proteins. The incubation time is 90 min at 30°C. RISC activity equals the amount of product divided by the sum of the amounts of product remaining substrate. The relative activity (Fold) was calculated by comparing the activity of each lane to that of lane 1. (C) The siRNA-initiated RISC assays were performed and analyzed as described in (B), except 25 nM siRNA was used and the incubation time was 45 min at 30°C. (D) RISC reactions were performed with 25 nM unmodified (lane 1) or biotinylated (lane 2) siRNA in S100, or with biotinylated siRNA in 10% PEG supernatant alone (lane 3) or in combination with 10 nM DCR-2 (lane 4), DCR-2/R2D2 (lane 5), or DCR-2/R2D2M (lane 6) recombinant proteins. The biotinylated siRNA and associated proteins were precipitated by streptavidin beads, resolved by SDS-PAGE, and Western blotted for AGO2 (12). 1924 26 SEPTEMBER 2003 VOL 301 SCIENCE www.sciencemag.org REPORTS to 6). This crosslinking was greatly diminished when the mutant DCR-2/R2D2M complex was used instead, although similar amounts of proteins were used (Fig. 4D). Interestingly, we observed two R2D2-siRNA crosslinked bands, one at 45 kD and another at 52 kD, which probably represented R2D2 proteins covalently linked to one or two siRNA strands (the mass of each 21-nt siRNA strand was 7 kD) (Fig. 4D, lane 5). This was confirmed by the downshift of both R2D2-siRNA crosslinked bands to the original position of 38 kD His6-R2D2 protein after RNase treatment (Fig. 4E). These results indicate that DCR-2 and R2D2 bind siRNA coordinately, which is dependent upon the dsRNA-binding domains of R2D2. On the basis of this finding, we hypothesized that R2D2 might be involved in facilitating siRNA loading onto RISC. To test this hypothesis, we separated the RISC activity from the Dicer activity in S100 by polyethylene glycol (PEG) precipitation and then combined the partially purified RISC with recombinant Dicer proteins to reconstitute the RNAi reaction. As shown in Fig. 5A, although the majority of siRNA-generating activity was precipitated by 10% PEG, a substantial amount of RISC remained in the supernatant, which could be activated by addition of siRNA for sequence-specific mRNA degradation. The wild-type DCR-2/ R2D2 complex was much more effective than DCR-2 alone or the mutant complex in promoting the dsRNA-initiated RISC activity in the 10% PEG supernatant (Fig. 5B). At 3 nM concentrations, wild-type DCR-2/R2D2 stimulated the RISC activity by more than sevenfold, whereas DCR-2 or DCR-2/R2D2M stimulated only by twofold (Fig. 5B; compare lanes 2, 4, 6, and 1). Interestingly, the mutant complex blocked the RISC activity at 10 nM concentration, possibly as a result of dominant-negative effects (Fig. 5B; compare lanes 7 and 1). Furthermore, a similar phenomenon was observed in the siRNA-initiated RISC assay (Fig. 5C). Thus, the DCR-2/R2D2 complex could enhance the siRNA- as well as the dsRNA-initiated RISC activities. We further showed that this enhancement was not simply because of siRNA stabilization by DCR-2/R2D2 by comparing the stability of radiolabeled siRNA in the RISC reactions described above (15). To confirm that DCR-2/R2D2 facilitates siRNA loading onto RISC, we followed the association between AGO2, an essential component of RISC, and a 3 -biotinylated siRNA by precipitation using streptavidin beads. The biotinylated siRNA was as active as unmodified siRNA in inducing RISC activities in S100 (15). However, streptavidin beads only precipitated AGO2 protein when biotinylated siRNA was used, suggesting that it was a specific interaction (Fig. 5D, lanes 1 and 2). We then performed RISC assays with the use of biotinylated siRNA in 10% PEG supernatant alone or in combination with recombinant DCR-2, DCR-2/R2D2, and DCR-2/R2D2M proteins. Consistently, more AGO2 proteins were detected in the biotinylated siRNA precipitates when wild-type DCR-2/ R2D2 complex was used instead of DCR-2 alone or the mutant complex. Together, our results indicate that DCR-2/R2D2 not only generates siRNA from dsRNA but also binds to nascent siRNA and facilitates its loading onto RISC. The latter activity is dependent on the dsRNAbinding domains of R2D2. This model is supported by previous studies in which the direction of dsRNA processing was limited to one end of the dsRNA molecule (11). In these experiments, if dsRNA was processed from the 5 to 3 direction of the sense strand, it would generate RISC that can mediate degradation of the sense but not antisense target mRNA, and vice versa. However, if synthetic siRNA was used instead, it produced RISC that could degrade either sense or antisense target mRNA. Therefore, the newly synthesized symmetric siRNA product must not become a free molecule once it is generated from dsRNA. Rather, the nascent siRNA is likely to be retained by DCR2/R2D2 in a fixed orientation determined by the direction of dsRNA processing such that only the antisense strand can become the guiding RNA for RISC. Both R2D2 and RDE-4 contain tandem dsRNA-binding domains, interact with Dicer, and are required for the RNAi pathway (13, 14). Thus, we conclude that R2D2 is homologous to RDE-4. RDE-4 also interacts with RDE-1, an AGO2 homolog and a RISC component. Therefore, we propose that R2D2 and RDE-4 play a similar role in bridging the initiation and effector steps of the Drosophila and C. elegans RNAi pathways, respectively. 1. A. Fire et al., Nature 391, 806 (1998). 2. M. K. Montgomery, S. Xu, A. Fire, Proc. Natl. Acad. Sci. U.S.A. 95, 15502 (1998). 3. T. Tuschl, P. D. Zamore, R. Lehmann, D. P. Bartel, P. A. Sharp, Genes Dev. 13, 3191 (1999). 4. G. J. Hannon, Nature 418, 244 (2002). 5. E. Bernstein, A. A. Caudy, S. M. Hammond, G. J. Hannon, Nature 409, 363 (2001). 6. P. D. Zamore, T. Tuschl, P. A. Sharp, D. P. Bartel, Cell 101, 25 (2000). 7. S. M. Elbashir, J. Martinez, A. Patkaniowska, W. Lendeckel, T. Tuschl, EMBO J. 20, 6877 (2001). 8. A. Nykanen, B. Haley, P. D. Zamore, Cell 107, 309 (2001). 9. S. M. Hammond, S. Boettcher, A. A. Caudy, R. Kobayashi, G. J. Hannon, Science 293, 1146 (2001). 10. S. M. Hammond, E. Bernstein, D. Beach, G. J. Hannon, Nature 404, 293 (2000). 11. S. M. Elbashir, W. Lendeckel, T. Tuschl, Genes Dev. 15, 188 (2001). 12. Materials and methods are available as supporting material on Science Online. 13. H. Tabara, E. Yigit, H. Siomi, C. C. Mello, Cell 109, 861 (2002). 14. A. Grishok, H. Tabara, C. C. Mello, Science 287, 2494 (2000). 15. Q. Liu et al., unpublished data. 16. S. Kalidas et al., in preparation. 17. G. Hutvagner, P. D. Zamore, Science 297, 2056 (2002); published online 1 August 2002; 10.1126/ science.1073827. 18. We thank G. Hannon, E. Bernstein, J. Abrams, and L. Ford (Ambion) for useful reagents; H. Shu for mass spectrometric analysis; J. Morse for embryo injections; and J. Goldstein, K. Lynch, J. Rutter, X. Jiang, L. Wang, and P. Mishra for critical reading of the manuscript. This work was supported by a high-risk grant from UT Southwestern Medical Center and by grants from the American Heart Association (EI0040029N) and NIH (DC02539). T.A.R. is in the medical scientist training program. Q.L. is a GlaxoSmithKline Fellow of the Damon Runyon Cancer Research Foundation. Supporting Online Material www.sciencemag.org/cgi/content/full/301/5641/1921/DC1 Materials and Methods Fig. S1 References and Notes 2 July 2003; accepted 13 August 2003 References and Notes Epidermal Viral Immunity Induced by CD8 Dendritic Cells But Not by Langerhans Cells Rhys S. Allan,1* Chris M. Smith,2,3* Gabrielle T. Belz,2,3* Allison L. van Lint,1 Linda M. Wakim,1 William R. Heath,2,3† Francis R. Carbone1† The classical paradigm for dendritic cell function derives from the study of Langerhans cells, which predominate within skin epidermis. After an encounter with foreign agents, Langerhans cells are thought to migrate to draining lymph nodes, where they initiate T cell priming. Contrary to this, we show here that infection of murine epidermis by herpes simplex virus did not result in the priming of virus-specific cytotoxic T lymphocytes by Langerhans cells. Rather, the priming response required a distinct CD8 dendritic cell subset. Thus, the traditional view of Langerhans cells in epidermal immunity needs to be revisited to accommodate a requirement for other dendritic cells in this response. Langerhans cells (LCs) represent the dominant dendritic cell (DC) subset within the skin epidermis (1). They are a mobile cell population capable of efficient antigen capture (2) and migration out of the skin on activation by a variety of stimuli (3–5). At the www.sciencemag.org SCIENCE VOL 301 26 SEPTEMBER 2003 1925


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subset: commoncrawl


text:
The opportunities were there, but the Barton Community College men's basketball team couldn't quite seal the deal on Wednesday night against Garden City.
Neither team ever held a decisive lead as Barton Community College's women's basketball team hosted Garden City.
Things went according to plan once the Western Athletic Conference Bowling Tournament finally got under way on Monday.
For Jason Parr, his quest to complete the 2014 Boston Marathon is a calling, inspired by the story of a courageous father-son duo.
The Great Bend High School boys' basketball team found their shot early, taking off in the first quarter to give themselves an early lead.
A hard-fought Western Athletic Conference game between Great Bend High School's girls' basketball team and Liberal came down the wire.
There is nothing that can prepare the Great Bend High School boys' swimming team for the Class 5-1A state swim meet.
With their senior year at high school waning, Kaylie and Kelsie Doll have made their decision about their future.
The decision was made on Friday.
The Kansas Jayhawk Community College Conference Western Division game was well in hand for the Barton women's basketball team.
When you're 14-for-20 from beyond the arc, it's hard to lose.
Even though the Cloud County Community College women's basketball team kept a strong hold on the game on Saturday night at Kirkman Activity Center, Barton was never very far behind.
Things started off slow for both teams, neither shooting better than 33 percent from the field in the first half.
With a healthy dose of 3-pointers in the first half, and Wyatt Bryant hitting his stride in the final eight minutes, Great Bend High School's boys' basketball team ran away with the game on Thursday night.
It wasn't long into the Hilltop Hoops Classic championship game that it looked like the Central Plains High School girls' basketball team was going to run away with it.
In the driver's seat.
Larned (4-3 overall, 0-1 district) at Hugoton (2-5, 0-1)
GARDEN CITY - At the end of the Western Athletic Conference volleyball tournament, it was host Garden City that sat atop of the standings.
SALINA - The Great Bend High School girls' golf team placed 10th at the Class 5A State Tournament on Monday.
Hoisington High School's football team took its first step into the Class 3A District 15 schedule.
It was a slow start for the Great Bend High School football team.
The Great Bend High School girls' tennis doubles team of Morgan Francis and Macy Moyers skated through Day 1 of the two-day Class 5A State Tournament.
Great Bend High School's first game into the Class 5A District 6 schedule couldn't have gone much better than it did.
It is a long road to the Class 5A State Tennis Tournament.
It is pretty common to hear that the high school football season doesn't really start until Week 7.
Holcomb (5-1) at Larned (4-2)
It was a big day for the Great Bend High School girls' golf team.
LIBERAL - It was their last chance at a Western Athletic Conference win.
SALINA - All season long, there has been two Great Bend High School girls' tennis players that have come out on top every time.
Central Plains 22, Little River 6


text:
Scenes From a Marriage
The long journey of the Manis marks Yehoshua's first sustained engagement with his own Sephardic roots, a development he ties to the death, in 1982, of his own father, an Orientalist and historian of Sephardic culture in Jerusalem. While the 1970s saw an attempt by militant Sephardic activists and intellectuals to articulate a Sephardic identity that had been deliberately repressed in the compulsive assimilation of Arab Jews who arrived in Ashkenazi-dominated Israel in the 1950s and '60s, Yehoshua vehemently resisted any Sephardic affiliation. His own Israeli identity was settled long beforehand, and the immigration, he attests, only sharpened his identification with the majority. "I fit in to Israel," he says, "like a French peasant to France."
Yehoshua's contempt for the diaspora is present in nearly all his novels, and it is a frequent subject in his nonfiction as well; he has called it "a disease" and "immoral," a "neurotic solution." While The Lover and A Late Divorce are set in motion by the return of a meddling diasporite, in The Liberated Bride Yehoshua gives us a lively caricature of Rivlin's sister-in-law Ofra and her husband Yo'el, who breeze through the country for a few days once or twice a year, all the while piously declaring their love for the homeland. "I'll still be Israeli," Yo'el solemnly vows, to Yehoshua's clear disgust, "even when there's no more Israel."
The question of Israel's future--the shape of the state and identity of its citizens--has always haunted Yehoshua's work, and it looms especially large in The Liberated Bride. Identity, with which he is so evidently concerned, finds itself in Israel at the mercy of geography, and the question of where to draw the lines, or to build the walls, is at root one of demography rather than territory: who can remain inside and who will be outside. Borders and boundaries--and their violation--structure the narrative of The Liberated Bride: Ofer's eagerness to penetrate the inner circle of the hotel, which leads him to discover the incestuous relationship of his father- and sister-in-law, is the original sin that sets the book in motion. Rivlin's reckless disregard for personal and national boundaries in his own search carries the story from there.
Obsessed with sickness and its diagnosis, The Liberated Bride gives us Carlo Tedeschi's hypochondria, Samaher's imagined illness and a terminal ailment Rivlin invents for himself to rouse the sympathy of Galya and her family. The book's panoply of professional Orientalists, meanwhile, are preoccupied with divining the mysteries of the "Arab soul," and the maladies said to lie within; Ephraim Akri, one of Rivlin's colleagues, repeatedly expounds a Bernard Lewis-style "theory of Arab failure" (he is even summoned to an American conference on Edward Said's Orientalism to counter the influence of Said's disciples, who are said to be "terrorizing the academic community").
Though it would be unfair to characterize The Liberated Bride as fundamentally an "Orientalist" novel, that spirit permeates it nevertheless. Like Tedeschi's wife, who needs the sickness of her partner, Rivlin, with his "long-craved intimacy with the Arabs," displays a complicated attachment to their afflictions. Indeed, as Rivlin testifies after Tedeschi's death, it was his anxiety over "our inability to understand the Arab mind" that gave rise to his hypochondria. The diagnosis of Israeli society, meanwhile, is left to others: It is telling that Professor Rivlin focuses on Algeria at the expense of the situation under his nose. ("When will you write something about us?" the Palestinians in Mansura inquire.) For the French in Algeria faced then a situation not unlike the Israelis today: forced to choose between withdrawal from Algeria and the integration of Algerians into France. In the end, of course, they got both.
"We have hooked our circulatory system to that of the Palestinians," Yehoshua said with great concern in a recent interview, and The Liberated Bride suggests that the solution to the spreading sickness is quarantine. Separation, between lovers and nations, is the road to liberation. But The Liberated Bride is, above all, an anxious book, and its plea for separation--between Ofer and Galya, Carlo and Hannah, Rashid and Samaher--cannot conceal a creeping unease about the efficacy of these liberations. It is a pitch-perfect evocation of the anxiety of liberal Israelis, and returns compulsively to a deeper and more troubling issue unlikely to be resolved by even the highest "security" wall: the identity and future of Israel's more than 1 million Palestinian citizens. The book's Arab characters, after all, are themselves Israeli citizens. They have a hybrid identity; Fu'ad, the hotel's maitre d', is an "Israeli Arab, who was not yet a Jew and no longer a true son of the desert." And this partial identity, like that of the Jews in the diaspora--"they are becoming our Jews," Yehoshua says of the Palestinians--cannot but represent an existential threat to Yehoshua's conception of Zionism, which is meant to redeem the catastrophe of the diaspora through a muscular and distinct definition of Israeliness and the separation of Jews from non-Jews.
Since the publication of his first stories nine years after the founding of the state, Yehoshua has sought to represent and articulate a vision of a consolidated but complex Israeli identity that might free his people from the traps of history. The rich depictions of the Israeli everyday found in A Late Divorce and Five Seasons accomplish this just as beautifully as the explicit interrogation to which he subjects the past in his masterpiece, Mr. Mani. But yesterday's traps are not necessarily today's, and while Yehoshua has soldiered on, history has refused to stand still; his ardor for a complete identity exists today in stark isolation from the scrambled Levantine reality. The failed policies of the last quarter-century--chief among them the settlement enterprise and its accompanying "separation" wall--have made separation less rather than more likely. With each year, each settlement, each suicide bombing and each "targeted killing," the mirrored peoples are entangled further in their bloody mise en abyme; the solutions to the past may be no help for the problems of the future.


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text:
River Forest Villager of the Year runner-up: Donna Carroll
President of Dominican University cites both 'positives and the pain'
By Devin Rose
Donna Carroll is a 57-year-old single woman—with more than 3,600 kids.
"Or none, depending upon your perspective," said the Dominican University president and a River Forest runner-up in Wednesday Journal's annual Villager of the Year.
Carroll is in her 18th year as president, and said she's proud of what Dominican has achieved in 2011.
In May, the school became the country's first university to enter Cuba for a study abroad program. There were two Fulbright Scholars named from the university. And the school took on a national leadership role in campus-based interfaith dialogue by welcoming Interfaith Youth Core Founder and White House appointee Eboo Patel as the year's Lund Gill Chair.
In addition, the school has hosted several other standing-room-only speakers and performers, Carroll said.
But as the school's head strategic planner, she said she is always focused on its next steps. Those include hiring more faculty members, increasing fundraising for student scholarships, advancing technology and constructing a new building on the Priory Campus at 7200 W. Division St.
Carroll said she's also excited about initiatives in the works through the school's Butler Children's Literature Center and Center for Global Peace through Commerce, and Dominican's collaboration with U.S. Bank on economic education.
But 2011 saw challenges, too. In this economy, the unpredictability of enrollment, philanthropy and state and federal funding has made leadership difficult, Carroll said.
She will continue to fundraise and build relationships throughout the next year. Being an engaged neighbor and resource is a big part of planning for the future, she said. The school working with the OPRF Community Leadership Program, OPRF Symphony and the River Forest Park District demonstrate the importance of partnerships, she said.
Looking back, Carroll said it was pretty gutsy of the Sinsinawa Dominican sisters to hire a young New York woman as their first lay president. But to this day, she says they are her greatest champions, "through all the positives and the pain."
Reader Comments
9 Comments - Add Your Comment
JSchutt from Worth
Posted: January 7th, 2012 12:48 PM
I am a DU Alum from the adult program that meets at the Priory. DU changed my life and I will forever be grateful. I live in the far south 'burbs and commuted to campus. DU faculty and students brings tax dollars to your community. I shopped and continue to do so in OPRF. My daughter, also a DU alum also does the same, still shopping and meeting friends at OPRF stores and restaurants. I often bring out of town guests to your tourist attractions. That's only part of what DU has done for OPRF
Tom from RF
Posted: January 7th, 2012 12:43 PM
RF PD is mentioned by Ms. Carroll as one of her "partners". Has she engaged her neighbor partner - the RF PD about the HIGH Rise dorm "strategic" plans for the Priory property that Ms. Carroll is focused on for her 10 year "GROWTH" plan?
Enuf is Enuf
Posted: January 5th, 2012 2:41 PM
Just because the gov set up a system to "help" those less fortunate, does not mean we should bend over backwards to a institution that is very well off, and has taken much more from the community, than put back in. Ms. Carroll cares more about her resume than the community. It is absurd. As to the percent, do not care if it is 53 or 1 - you decide. BTW, they only own a part of the land, believe the rest is owned by RF Park District?
John Butch Murtagh from Oak Park, Illinois Facebook Verified
Posted: January 5th, 2012 11:36 AM
Wow, a Catholic Sertus!
53 Percenter from Oak Park
Posted: January 5th, 2012 11:00 AM
Enuf, what is your problem with another building at the Priory? It's DU's land to do with as they wish. It's also a huge parcel that could easily absorb a building with zero impact on the area. And please get over yourself. No university, public or private, pays property taxes. Like it or not, that's how we do things in the US. DU is an extraordinary civic asset. And can we please stop this 1% Occupy nonsense too? The only important % is the 53% of us who pay income taxes to support the 100%.
Enuf is Enuf
Posted: January 5th, 2012 10:55 AM
We pay our taxes, thank you! Also, no reason to post under two names @Enough. Any non profit must show how it benefits the community. The traffic and the crowd brought in by Dominican - has gotten too large. We have reached the proverbial "tipping point" Dominican wants to put up 10 story dorm at the Priory. We will see how the Priory Neighbors and the RF community will react - once the public hearings begin.
Enough already
Posted: January 5th, 2012 9:56 AM
Enuf pay your taxes!!
Posted: January 5th, 2012 8:37 AM
Dominican already owns the Priory, perhaps? and they are already not paying taxes on the property and I did not see that they were asking for any money from the village, so what does it matter?
Enuf is Enuf
Posted: January 5th, 2012 8:22 AM
A new building at Priory - WHY?? Please Explain Ms. Carroll and WJ, please dig into this. This needs to stop now, letting them take over RF. This is a residential community, AND WE PAY A LOT OF taxes. Dominican - PAYS Nothing. Dominican has enough buildings already. Neighbors - let's get ready to NOT let this happen. This is the 1% railing against the 99%
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text:
If it feels like the combine just ended for you imagine being a NFL scout right now with a full month of non stop travel ahead of you. With the combine officially wrapping on Tuesday the NFL Pro Day schedule kicks off on Friday with a pair of FBS schools that will get to show off the best prospects in front of a contingent of NFL scouts. Both Purdue and UTEP elected to have their Pro Days early this year setting them apart from what is a very packed schedule for the rest of March. (more…)
Posts Tagged 'Kawann Short'
Pro Day Primer: Purdue and UTEP (3/1)
Thursday, February 28th, 2013
Prospect Preview: Heart of Dallas Bowl
Tuesday, January 1st, 2013
BBD Writer Tony Wiltshire
After a disappointing season, Oklahoma State (7-5) will meet Purdue (6-6) in the 2012 Heart of Dallas Bowl. The teams last met in 1997 in the Alamo Bowl, and it should be interesting to see if the Boilermakers can limit the explosive Oklahoma State offense. There are players on both sides of the ball that will soon extend their careers in the NFL. (more…)
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The Denver Broncos enter the 2014-2015 season with a heap of expectation. The largest concern of the team however is if they can keep two staples of the record-breaking offense in Julius Thomas and Demaryius Thomas. These two players are coming into their own and being widely considered as one of the best in the game at their respective position. The bad thing, however is can the Broncos keep both these players given that both of their contracts are up at the end of the year.
It can be done, but it's risky. John Elway has gone all in for the Broncos and the expiring contracts this year include Demaryius Thomas, Julius Thomas, Terrance Knighton, Chris Harris, and many more. There is no question in my mind that Demaryius Thomas is a top five receiver in the league, and he deserves to be paid as much. The question lies with the other Thomas.
Julius Thomas is emerging to be a top ten tight end in this league and there is no doubt that teams will be looking around for that franchise TE, and that is Julius. The Broncos will have a good amount of salary cap to play with at the end of the year with so many contracts expiring, so Julius should get top TE money. The potential problem that lies here is if Julius is going to try to do a Jimmy Graham and attempt to classify himself as a WR and attempt to get top WR money, which is around $10 Million a year as opposed to top TE money which is around $7 Million dollars a year.
Demaryius Thomas is going to get paid. It's more than likely to come after the Super Bowl as we are little over a week away from the opening game of the season, it makes sense to do negotiations after the season so that it doesn't distract the player or the team. The question is Julius. You should fully expect that both Thomas' will be back in Denver because Julius isn't the kind of guy to try to change his position just to get more money. Both players should get max extensions and be Broncos for life.


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Yes, these are dire political times. Many who optimistically hoped for real change have spent nearly five years under the cold downpour of political reality. Here at CounterPunch we've always aimed to tell it like it is, without illusions or despair. That's why so many of you have found a refuge at CounterPunch and made us your homepage. You tell us that you love CounterPunch because the quality of the writing you find here in the original articles we offer every day and because we never flinch under fire. We appreciate the support and are prepared for the fierce battles to come.
Unlike other outfits, we don't hit you up for money every month … or even every quarter. We ask only once a year. But when we ask, we mean it.
CounterPunch's website is supported almost entirely by subscribers to the print edition of our magazine. We aren't on the receiving end of six-figure grants from big foundations. George Soros doesn't have us on retainer. We don't sell tickets on cruise liners. We don't clog our site with deceptive corporate ads.
The continued existence of CounterPunch depends solely on the support and dedication of our readers. We know there are a lot of you. We get thousands of emails from you every day. Our website receives millions of hits and nearly 100,000 readers each day. And we don't charge you a dime.
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It's Over, Thank God
It's finally over. I had grown weary of having presidential campaign ads interrupting the commercials. As much as I despise commercials interrupting programs, at least they have something concrete to sell like a car or a beer. This seemingly eternal campaign seemed to begin years ago. Indeed, by late October, more than 915,000 campaign messages had appeared on U.S. TV sets – a record that belongs in the Guinness Book of Stupid Records, especially if you analyze the content of the campaign messages: Vote for Beaver's father (1950s TV), who will fix the economy his big shot investor friends helped break, or vote for the cool guy who only disappointed you.
God forbid, literally, that the candidates should have discussed what scientists repeatedly warn us about: the threats posed by climate change to future human life if we don't take drastic steps to stop greenhouse gas emissions. Romney even pushed the notion of burning more coal (he claimed it's clean) and drilling (how thrilling!) for more oil. Obama had done little to curtail the big burners and neither of the two had taken any leadership on the climate change crisis. Indeed, both candidates supported nuclear power expansion. Romney even wanted to get rid of FEMA. Oops! A major sensitivity slip as Hurricane Sandy hit! Obama said "no red tape" in getting help to people.
Obama also said he wanted to finish the job he undertook in 2008, rebuilding the country. How can he do that and continue to pay for the rising costs of the U.S. empire? Romney complained Obama wasn't spending enough on the military (horses and bayonets cost more these days), and conjured up foreign enemies (Russia and China) whose militaries do not threaten us. Obama insisted that real enemies do exist and he had routinely murdered them with Seals or drones in remote places. Pakistanis, for example, now fear going to weddings lest they get mistaken at Drone headquarters for people going to an Al Qaeda convention.
What didn't get discussed? The millions of Americans who live in deep insecurity and downright poverty, themes that don't relate to the ambitions of the aspirants who blather about pseudo patriotic nonsense, but didn't mention the 2-plus million people in U.S. prisons.
Both candidates also shared close friends among the Wall Street gamblers who had helped precipitate the economic crash. These economy-busting thieves continued to evade arrest, but in Colorado police arrested 210,000 people between 1986 and 2010 for marijuana possession. Colorado's marijuana possession laws got enforced in a colorful way. Young whites are more likely to use marijuana than any other ethnic group, but African Americans and Latinos made up a disproportionate percentage of those arrested for marijuana possession. An African-American is more than three times as likely to be arrested as the Caucasian, although African-Americans make up less than 4 percent of the state's population. They account for over 10 percent of the marijuana arrests. A new FBI report states that police arrest someone for marijuana possession every 42 seconds.
The Bureau gives no details on the frequency of banker arrests, but make your own guesses. The candidates did not call for the arrest and prosecution of the muckety mucks on Wall Street for leading the U.S. economy into its sub-prime mortgage calamity. The contenders showed no interest in stopping drone murders or warrantless surveillance, indefinite detention, or racial profiling in the unending war against terrorism.
U.S. reality has grown distant from the babble of self-serving "truths" uttered in campaign rhetoric, where eight Senate candidates agreed that God intended pregnancies resulting from rape. And Romney actually endorsed some of these mountebanks. The fact that abortion has become the lead issue on several political campaigns should shock most of the world's population. And don't forget: Jesus told us: "Arm yourselves."
As millions of Americans suffered eviction as a result of home foreclosure, and countless millions more lost jobs and health plans, while many wait anxiously each day to see if their jobs would remain, the aspiring politicians talked about our great and powerful country, as if such language proved their presidential characters. It's hard not to note how they avoided issues that grip the guts of a people caught in an economic undertow they cannot control. Romney promised to create jobs, mystically and magically, but offered no plan other than cutting rich people's taxes. Obama did not return to FDR's ideas of creating large public jobs' programs to rebuild the infrastructure.
Mitt Romney displayed, on a secretly recorded video, his for 47% of U.S. citizens, not illogical considering Bain Capital, his private equity firm, enticed reactionary Salvadoran financiers, linked to government-backed death squads, to fund his operation. Paul Ryan's budgetary ideas offered to undo what's left of the New Deal and Great Society. Neither candidate refused to take huge campaign contributions from people and organizations. The candidates seemed satisfied with the system of raising large sums of money from super rich people – the essence of U.S. democracy.
On humanitarian and civil liberties questions, neither candidate showed interest in ending the death penalty or reducing the 5,000-plus U.S. nuclear warheads. Obama's pledge to close the Guantanamo prison and torture center further raised liberal eyebrows, especially after he didn't do it. But he did brag about killing Osama bin Laden. And Romney didn't criticize Obama for not trying to bring bin Laden to trial instead of slaying him. Both candidates claimed toughness (bombing on the table) on Iran and boasted that their lips were disgustingly close to Israeli butt. A truly tempting reason to vote for them!
Obama will appoint better judges, more serious people to government agencies like the EPA and NLRB, and not endanger women's rights. So, I voted for Obama=Bad, rather than Romney=worse.
On Wednesday they told me we'd won. I wonder.
Saul Landau's FIDEL and WILL THE REAL TERRORIST PLEASE STAND UP are available as dvds from cinelibrestore.com.


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Revlon • ColorStay Makeup with SoftFlex SPF 15 Normal/Dry Skin • Liquid
(483 reviews)
|Average Rating.
|Would buy this product again.
jadeseymour on 3/24/2014 3:28:00 PM more reviews by jadeseymour
Age: 18 & Under Skin: Dry, Fair Hair: Brunette Eyes: Hazel
pros: -long lasting -lightest shade works with skin tone -medium-full coverage -covers my moderate rosacea and acne -doesn't make me break out -not extremely pricey -not easily breakable -doesn't cling to dry areas cons: -no pump -container can be misleading on how much product is in bottle -lightest shade is on the pink side overall this foundation it great and dispute my lack of love for the packaging it is still my HG foundation and better then any other foundation from the drugstore . if you are hesitant to try it don't be you won't be disappointed ;) will repurchase when I run out and will continue to .
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
CharacterofLife on 3/22/2014 3:51:00 PM more reviews by CharacterofLife
Age: 30-35 Skin: Normal, Fair-Medium, Warm Hair: Brunette Eyes: Blue
This is my favorite"foundation" foundation. I've been loving Physicians Formula's BB & CC creams a lot but decided to try a real foundation again. This one is the best I've found so far. I have light/medium olive skin. The type that has an apparent light green hue too. Foundations that are too punk, yellow, or orange make the green stand out. I do not think I am an alien nor an alien hybrid, well the aliens have not informed if such.... Yet.
This formula dries at a medium pace. I use Olay Sensitive SPF 15 under it. It allows enough time to spread then it sets well without being drying my skin out. The color fresh beige goes so well with my skin I can almost claim it to be a perfect match... For an alien in Earth that is.
It does make my fine lines a bit apparent, but the coverage is so wonderful I'll keep it!
It has great staying power, doesn't dry my skin out yet seems to help keep my t zone less oily.
My only three wishes would be that it came in a pump, down didn't emphasis my lines, and was came with a Jeremy Renner. Why? Because he is so sexy of course. He is sexy throughout the galaxy.
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
elenidem on 3/15/2014 1:00:00 PM more reviews by elenidem
Age: 18 & Under Skin: Other Hair: Other Eyes: Other
I heard so many awesome things about this foundation, people said it would cover spots and blemishes perfectly and not look too cakey, so whats not to like?
The colour selection is very good and im in the lightest one, Ivory, which is a bit too pale for me compared to the bourjois healthy mix one.
However, it dries VERY quickly so you need to move fast when you are blending and it does leave brush marks so blend VERY CAREFULLY.
So I put this foundation on hoping it would do all these things for my skin and it just didnt do it. It looked so cakey on my face and made every little hair or bump on my skin noticable and as for covering my blemishes, it just made them worse. From far away my skin looked pretty good but face to face it looks so awful and i feel so self conscious when wearing it, especially at school! And for being in the dry formulation it just made my dry patches stand out especially on my nose it looks so bad i actually wanted to cry.
I am so disappointed because really wanted it to work for me. I dont know if im putting it on right or not because everyone on youtube seems to think this is a holy grail foundation.
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loucat on 3/14/2014 10:36:00 PM more reviews by loucat
Age: 44-55 Skin: Dry, Fair-Medium, Cool Hair: Brunette, Wavy, Fine Eyes: Brown
I'm 50 yrs old and buy the normal/dry in the color sand beige, it stays on all day, u can see a little bit of my fine lines but not to bad, this is the best price at walmart, i use this for everyday wear and for going out i use estee lauders double wear in the color pebbles . i did notice the bottle top starts cracking and breaking towards the end of the makeup but other than that its well worth the price
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*TOP REVIEWER* Walko on 3/14/2014 6:57:00 AM more reviews by Walko
Age: 56 & Over Skin: Combination, Medium Hair: Other Eyes: Brown
This has got to be one of the best foundations out there. I've used this for years on and off.Recently I've been using EL double wear stay in place make up spf 10. EL is good but colorstay IS comparable and in some ways favourable.For example if it's extra sun protection you're after , Revlon wins hands down. It contains both titanium dioxide and zinc oxide in it's sunscreen,has a higher percetage of sunscreen ingredients and has spf 15.I don't rely on any foundation as a sunscreen alone , I use a separate sunscreen underneath, otherwise you'd need to cake on the make up to be protected. This has no chemical sunscreens in it which are often the cause of stinging eyes.Colorstay has excellent coverage, great staying power.It's semi matte I would say it's not that mask matte, I'm 57, I don't need that.I find it to be quite natura looking, even though it's full coverage, depends on the individual I suppose. I use the shade sand beige and it's excellent for my NW 15 combination skin..This is a top pick on beautypedia ,so it's not full of irritants.This must be removed thoroughly,it can be be hard to remove.But any make up has to be removed properly.I think a lot of "breakouts" is just bad skin care.This foundation has been around for years, but if you've not tried it yet, I'd say it's sure worth a shot.
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BeautyAddict62 on 2/20/2014 1:19:00 PM more reviews by BeautyAddict62
Age: 19-24 Skin: Combination, Fair, Cool Hair: Brunette, Wavy, Medium Eyes: Green
(Not Softflex, just the normal version)
I purchased this Colourstay foundation after hearing it was a good dupe for Estee Lauder Double wear......hmmmmm it is a good enough foundation.
I have it in Ivory which is a perfect shade for my pale skin however it is extremely drying. It has a tendency to cling to dry patches around my nose, cheeks & forehead and makes them even drier! I would advise to moisturize your skin very well before applying. Overall, it is not my favourite drugstore foundation but it is good if you're looking for heavier coverage.
*Fantastic colour match to my skin, absolutely no orange tinge whatsoever.
*Nice, sleek & sturdy packaging.
*Lasts all day without getting oily.
*If you have very oily skin, no need to worry as this is extremely matte.
*Great coverage, really hides my red areas, freckles & pores.
*Pricey for a drugstore foundation, I paid £12.49 for it.
*VERY drying on skin, despite the fact its meant to be for 'normal/dry' skin.
*Leaves a very chalky, matte finish which is great if you suffer from oily skin but it can look very un-natural & flat on normal skin.
*Can get cakey if applied with a heavy hand.
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
dtormon on 2/14/2014 1:51:00 PM more reviews by dtormon
Age: 19-24 Skin: Dry, Medium, Warm Hair: Brown, Wavy Eyes: Black
Great coverage for decently-priced drugstore foundation! It layers easily and has good lasting power -almost 8 hours. My only complaints are that the glass-top packaging isn't easy to keep clean or sanitary, and the finish almost makes me look chalky (and I do moisturize well in the winter).
Was this review helpful to you? Yes No
sS1221R on 2/10/2014 12:43:00 AM more reviews by sS1221R
Age: 18 & Under Skin: Dry, Fair-Medium Hair: Blond Eyes: Green
bought this foundation after hearing the hype and needing a new winter foundation after becoming too light for my others. found my perfect shade and this foundation has great coverage. also stays FOREVER with powder. i have problems getting it off if i also use a setting spray. lasts through workouts, concerts, raves, school, etc. only downside is it gives me a weird finish. almost makes my skin look drier. other than that, amazing foundation
Was this review helpful to you? Yes No
*TOP REVIEWER* miszxlinda on 2/9/2014 3:22:00 PM more reviews by miszxlinda
Age: 25-29 Skin: Sensitive, Medium, Cool Hair: Black, Wavy, Fine Eyes: Brown
I really wanted to try a new foundation from the drugstore and decided to finally give this a go. I am assuming I picked up a newer version \ formula of this as mine claims to be 24 hours... Was it 16 before? Well anyway, I am in love with the product. I work 12 hour shifts and most foundations I've tried don't seem to last. I have dry skin but will get oily towards the end of the day and often will see patches of my natural skin peeking through the day. But not with this! This formula is thick, a little difficult getting out of the bottle but blends into the skin quite nicely. Although thick, I didn't feel the need to use concealer... This seemed to have FULL coverage & hide my blemishes very well. I have NC 30 skin right now and picked up the shade 240 medium beige. Looked pale when I had it on first thing in the morning but it oxidizes and matches pretty decent! Total love!
Was this review helpful to you? Yes No
Britini250 on 2/1/2014 10:48:00 PM more reviews by Britini250
Age: 19-24 Skin: Acne-prone, Fair Hair: Blond Eyes: Brown
Best foundation for the money! Gives great coverage and lasts a long time.
Top Rated Liquid
Sisley Phyto Tient Eclat Oil-Free Foundation34 reviews


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Pics of basic dual battery setup.
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Album: Basic Dual Battery setup
Pics of basic dual battery setup.
Basic Dual Battery setup
Updated Jan 7, 2013
XenGallery by sonnb


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"One could say that photography, a certain photography is a very erotic practice: this way of almost fondling the subject, encircling it, modifying its attitudes, but more than anything maintaining a distance from the idea that the camera is magic and infernal, that the disposition of the lenses and the machinery make of it an object of extreme power, like the Bibi Fricotin mind-reading glasses, or the glasses that undresses salacious advertisements: this face, this body that I have before me, in the duration of the pose, the focus, the shutter release, I am with it as though I were probing it, it is a second of truth or lie which will produce itself, but something will appear, something will reveal itself, something will betray itself. I will know something more, I will imprison it, and it will be like evidence. The secret of the other shall be my secret. And this face which stares at me can very well decompose: it is already dead."
Hervé Guibert


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Talented young prop Aaron Woods has today been named the Wests Tigers Player of the Year for 2012.
Woods enjoyed a superb second season in the NRL, playing all 24 matches for Wests Tigers.
In stepping up to another level, Woods earned a City Origin jersey and Prime Minister Like
Aaron Woods named Tigers player of the year
- by Wests Tigers
- September 28 2012 7:19PM
Talented young prop Aaron Woods has today been named the Wests Tigers Player of the Year for 2012.
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To access the full Church Times online archive please log in above with your Church Times user email & password
Posted: 11 Jun 2008 @ 00:00
FOR ALL the jargon words about social welfare provision, the categories used by Christ to describe those in need of care (in Matthew 25) still hold good: naked, sick, in prison. The duty of the Christian is unambiguous: he or she is to clothe, care for, and visit those who suffer. In every generation there will be those drawn to a faith that concentrates on praise and worship; and others whose passion is to make the gospel known more widely. But a key part of the good news is that love, the fuel of God's kingdom, is already released into the world through the work of the Holy Spirit. Nor is there any doubt among the New Testament writers that those who do the will of God by caring for the poor, sick, and imprisoned — for whatever reason, whether believers or not — are, indeed, doing the will of God.
This is an important consideration when the Church looks at how it cares for the poor in the 21st century. One of the key findings of Moral, But No Compass, described here by Bishop Lowe, is that the Church has not really looked at how it cares for the poor. It is, perhaps, unfair to expect the Government to have a decent grasp of the Church's welfare activities when there is no overall strategy, no central gathering of statistics, no co-ordination between dioceses and independent agencies, and, most important of all, no consistency about what the Church can provide from one part of the country to another.
Of course, the last thing the Church wants is to set up a bureaucracy in parallel to the state's; but neither would it want to tie itself to a state organisation and lose its freedom to act swiftly in response to local need. The Church cannot offer to fund teams of professional carers, but neither can it promise to provide a range of services beyond the ability and expertise of its volunteer workers. Examples can be found of all methods of organisation and co-operation, but, as the Von Hügel Institute report found, the Church's welfare work is inevitably patchy. This being the case, it is essential that the Church's patchwork pieces are fixed to those provided by other agencies, either faith-based, charitable, or state. Without this, yet more holes in the fabric will appear for the poor and disadvantaged to fall through.
Church schools are a good model for other areas of social work. Clearly, no other field of social provision has such a huge legacy from earlier centuries (predating secular provision); none the less, many lessons can be learnt from the church-school sector about the balance between independence and reliance on the state. Until recently, it might also have been argued that, in this instance at least, the Church's value was recognised by the Government. In the past few months, however, it seems that, even here, there are figures in the Government who are not comfortable with partnership.
Esgobeath Bangor
The Diocese of Bangor
Eglwys sy'n Dysgu A Learning Church
The Bishop of Bangor wishes to appoint a Priest in Charge
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Free Agent D'Qwell Jackson Visits Broncos Headquarters
March 4, 2014 6:05 AM
Broncos CentralBuy Broncos Tickets Shop for Broncos Gear NFL Scoreboard NFL Standings Team STATS Team Schedule Team Roster Team Injuries
DENVER (CBS4) – The Denver Broncos visited with former Cleveland Browns linebacker D'Qwell Jackson on Monday.
The hope was to get the veteran signed before he left Denver. It didn't work out that way.
Jackson has two more teams to visit and expects to make a decision by the end of the week.
Jackson was drafted by the Browns in 2006 and made 96 starts in eight seasons with the team. They cut him after the season ended.


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Md. air station settles hazardous waste violations
The U.S. Navy has agreed to pay $38,500 to settle alleged hazardous waste violations at the Naval Air Station Patuxent River.
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Navy settles Va. hazardous waste allegations
The U.S. Navy has settled allegations that it violated hazardous waste and underground storage tank regulations at a Virginia Beach facility.
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A Baltimore County couple helped a wounded veteran and his fiancée travel home for the holidays.
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Dick Jones won the North State Regional 1,000 Yard Championship at Camp Butner, N.C., two consecutive years. The competitive shooter relays some of the tricks, tips and considerations of high-performance long-range shooting.
long-distance shooting
Bushnell's Bryan Morgan take No. 1 at Sniper's Hide Cup
Team Bushnell Tactical's Bryan Morgan came out top dog in one of the more testing long-distance shooting competition – the Sniper's Hide Cup.
Caldwell's Digital Wind Meter Helps Shooters Avoid Getting Blown Away
Wind can be one of the most challenging environmental factors facing shooters. Caldwell has dialed down the breeze's effects a bit with the introduction of its new wind meter.


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Card Table XtenderItem # HN-OH001
- Increases seating to six for dinner, or up to eight for a card game
- 2 side straps with closures allow secure attachment to card table legs with either side facing up
- Upholstery padded leather-grain vinyl top with a woven polyester underside to protect the table surface from scratches
- Cleans with damp cloth
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Comments about Card Table Xtender:
I needed to enlarge a 42 in round table. This 54 inch extender will work nicely for the occasional need. The trick was to figure out how to keep it from sliding on the glass, as the attached tabs are designed for card table legs, and don't work with my table. I was able to fashion a way to take advantage of the felt underside of the extender to keep it from sliding on the glass.
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Excellent product. Cranberry color was perfect and we have found the extender to be very useful in turning our card table into a 6-8 person dining table. Highly recommend!!!
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Added Jul 12, 2012
by carmencitab
Brittany, France
It's a men's shirt. Nothing special, except I really paid attention to the fine details.
My husband is very happy.
Jul 15, 2012, 07.12 PMby elguanche
Where the hell did you get this buttons???
I loooove it!
Jul 16, 2012, 07.21 AMby carmencitab
At a local sewing place a few months ago. I can check if I can find some more for you if you want.
Jul 15, 2012, 09.16 AMby carmencitab
Thank you all!
Jul 13, 2012, 07.17 PMby pamchar
J'adore les boutons!
Jul 13, 2012, 01.42 PMby alexus1325
Great job matching the plaid! Looks like a great fit on him, and those eagle buttons are just awesome!
Jul 12, 2012, 09.48 PMby wildcatspow
Nicely done! This is a sharp-looking shirt!
Jul 12, 2012, 08.49 PMby ctharp
Great looking shirt! You did great with the plaid….many kudos to you.
Jul 12, 2012, 02.33 PMby deChez
Beautiful work…and it fits him nicely.
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First name content excerpted from The Complete Idiot's Guide to 30,000 Baby Names © 2005 by
Marcia Layton Turner.
this book online or call 1-800-253-6476.
Last name content copyright © 2006 Patrick Hanks.
First name origin & meaning:
Vietnamese: Full understanding, knowledge
First name variations: Mina, Minnah, Minette, Minetta, Minda, Myna, Mindy, Minna, Wiletta, Wilette, Wilhelmine, Willa, Willimina, Wilhelmina, Wilma, Wilhelmina, Wilhemine, Wihelma, Willette, Wilmette, Wyla, Willie, Willy, Will, Mina, Minnie, Minny, Billie, Billy, Helma, Vilma, Guillelmine, Guillemette, Vilhelmina, Willa, Bell, Hallem, Eilon, Elan, Ilon, Ilan, Kalan, Lemar, Lemarr, Larmar, La Monte, Salomi, Solomon, Schlomo, Shlomo, Soloman, Salomon, Salmon, Sholem, Shalom, Solom, Saloman, Sol, Soloman, Solomon
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The Population Health Management community showcases the latest news and innovative trends on wellness programs employers are executing to help maintain or improve the overall health of each individual employee as well as the entire population, cutting health care costs. This could range from sponsored fun-run, smoke cessation, biometric testing, health and performance, etc.
The reasons for ensuring a healthy workforce seem clear enough: Not only is it the right thing to do, it's also good for the bottom line. Employees who engage actively in effective wellness plans are more productive, recover faster and miss less work. Moreover, a focus on wellness helps drive
Engagement has been thrown around as the metric of choice for measuring employee health.
I first cut my teeth as a wellness program coordinator during my internship at the Mattel toy company. It was truly the dark ages. No Web. No smartphones. No social media. We scanned HRA questionnaires through optical readers and gave out printed reports. We delivered classroom educational programs and one-one-one
Technology, when applied properly and wisely, can improve the way we live and work. It can help us make better use of staff time and resources. It can make people productive and processes more cost effective. It can also help us connect and communicate with people and is dramatically changing
07/22/2014 - Private exchanges are coming, but how will the transition affect the various stakeholders in employee...
07/22/2014 - When it comes to making an important decision, a significant investment or even beginning a...
07/16/2014 - On this week's program, your hosts Brent Macy and Doug Field will be exploring issues...
07/09/2014 - We hope all of our listeners and supporters had a fun and safe July 4...
Former Extend Health CEO and Towers Watson executive with deep sales and marketing expertise set to position SeeChange Health for growth in evolving market
Novu today announced the results of its Diabetes Prevention Program (DPP) pilot program.
Employee engagement technology provider is first investment for Oak HC/FT Partners' $500 million Healthcare and FinTech fund
In the past decade, corporations have implemented several different cost‐cutting measures and wellness initiatives: premium differentials, higher deductibles, wellness coaching, disease management, onsite clinics, HRAs and education. Still it has been an uphill and largely losing battle to measure the results of these programs. Many
Population health management programs are now solidifying into successful hospital initiatives with real results. Along with this trend, health systems are recognizing that viewing patients as consumemers is vital to creating a point of differencein their marketplace -- as health care becomes a "buy" decision.
This Consensus Statement was prepared by a Joint Committee of the Health Enhancement Research Organization (HERO), American College of Occupational and Environmental Medicine (ACOEM), American Cancer Society (ACS) and American Cancer Society Cancer Action Network (ACSCAN), American Diabetes Association, and American Heart Association (AHA). This


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Here's what I'm hearing on the Star Wars writers: not courting but that deals are closed. And they will also be producers. #StarWars— Borys Kit (@Borys_Kit) November 21, 2012
Academy Award Winner Adam Davidson to Direct The Walking Dead Companion Series Pilot
Full 16-Minute Quantum Break Gameplay Demo Online
The Nolans Do Not Have a Hand in the Upcoming DC Cinematic Universe
Paul Greengrass to Direct Berlin Wall Escape Tale The Tunnels
The Lord of the Rings' Billy Boyd Performs End Title Song for The Hobbit
Paul Giamatti and Damian Lewis to Star in Showtime's Billions
Warner Bros. to Remake German Blockbuster, Who Am I
Warner Bros. Lands Will Eubank's World Breakers
Sebastian Stan Joins Meryl Streep in Ricki and the Flash
Exclusive Clip from Before I Go to Sleep