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PUF (Pumilio/FBF) RNA-binding proteins and Argonaute (Ago) miRNA-binding proteins regulate mRNAs post-transcriptionally, each acting through similar yet distinct mechanisms. Here, we report that PUF and Ago proteins can also function together in a complex with a core translation elongation factor, eEF1A, to repress translation elongation. Both nematode and mammalian PUF/Ago/eEF1A complexes were identified, using co-immunoprecipitation and recombinant protein assays. Nematode CSR-1 (Ago) promotes repression of FBF (PUF) target mRNAs in in vivo assays, and the FBF-1/CSR-1 heterodimer inhibits EFT-3 (eEF1A) GTPase activity in vitro. Mammalian PUM2/Ago/eEF1A inhibits translation of nonadenylated and polyadenylated reporter mRNAs in vitro. This repression occurs after translation initiation and leads to ribosome accumulation within the open reading frame, roughly at the site where the nascent polypeptide emerges from the ribosomal exit tunnel. Together, these data suggest that a conserved PUF/Ago/eEF1A complex attenuates translation elongation.
Translational regulation impacts metazoan development, learning and human disease1–3. Of the many RNA-binding proteins regulating mRNA stability, localization and translation, the two classes relevant to this work are the PUF and Ago families. PUF proteins regulate a broad spectrum of mRNAs to maintain stem cells, among other functions1,2. Ago proteins bind small RNAs, most notably miRNAs, and act in many biological contexts, including stem cells (reviewed in 3).
PUF proteins regulate mRNA expression in virtually all eukaryotes4. One conserved mechanism, which has been established for Saccharomyces cerevisiae, Caenorhabditis elegans and human PUF proteins, relies on the direct recruitment of the Ccr4-Not deadenylase complex to target mRNAs, resulting in a shorter poly(A)-tail length and either mRNA instability or translational repression5,6. PUF proteins in yeast and flies also use deadenylation-independent mechanisms, although the specifics of these mechanisms remain largely unknown7,8.
Ago proteins function within the miRISC complex to control target mRNA stability and translation. This complex controls deadenylation and stability of its target mRNAs9–11, but its mechanism to control translation has garnered controversy. Best documented is its regulation of translation initiation9–13. However, a mechanism affecting translation elongation also appears to exist since target mRNAs can be bound to polyribosomes14. One model is that miRISC promotes ribosomal drop-off during translation elongation15. Yet the specific mechanism by which miRISC, and hence Ago family members, inhibits translation elongation is not understood.
Here, we report that PUF and Ago proteins form an inhibitory complex with eEF1A, a GTPase required for translation elongation. FBF-1 (a C. elegans PUF protein) binds CSR-1 (a C. elegans Ago family member) in vitro and in vivo, and csr-1 depletion leads to increased expression of FBF target mRNAs. The FBF-1/CSR-1 heterodimer forms a complex with EFT-3 (C. elegans eEF1A), and this FBF-1/CSR-1/EFT-3 ternary complex has inhibited GTPase activity. Importantly, the PUF/Ago/eEF1A complex is conserved: human PUM2 (a PUF protein) associates with human AGO proteins in vivo and with eEF1A. Wild-type human PUM2 inhibits translation of both nonadenylated and polyadenylated mRNAs in rabbit reticulocyte lysate; however, PUM2 mutants that cannot form the PUM2/Ago/eEF1A complex or that cannot bind RNA are severely compromised for translation repression. Mechanistically, PUM2/Ago/eEF1A represses translation during elongation with ribosomes accumulating ~100–140 nts after the AUG within the open-reading frame (ORF). We propose a model in which PUF and AGO proteins complex with eEF1A to inhibit its GTPase activity and attenuate translation elongation.
To explore the molecular mechanisms of PUF protein function, we sought proteins that interact with two nearly identical C. elegans PUF proteins, FBF-1 and FBF-2, known collectively as FBF16. Previous yeast two-hybrid screens identified several FBF partners, but no components of the translation machinery17,18. Here, we used recombinant, full-length GST-FBF-1 and GST-FBF-2 to select proteins from C. elegans lysates and identify interacting proteins by mass spectrometry (Supplementary Fig. 1, Supplementary Table 1). Known FBF-interacting proteins, CPB-1 and NOS-3 (refs. 17,18), were detected in addition to previously unknown putative partners CSR-1, one of 27 C. elegans Ago family members19,20, and EFT-3, the C. elegans homolog of translation elongation factor 1A, eEF1A21.
To directly test whether FBF-1 interacts with CSR-1 and EFT-3 in vivo, we immunoprecipitated tagged FBF-1 (ref. 22) from C. elegans lysate and probed for associated proteins. CSR-1 was specifically enriched, and this interaction was RNA-independent (Fig. 1a), indicating that FBF-1 and CSR-1 form a protein complex. By contrast, Dicer-related helicase (DRH-3), which is required to produce the 22-nt RNAs bound by CSR-1 (refs. 23,24), did not co-immunoprecipitate (IP) with FBF-1 (Fig. 1a), demonstrating that FBF-1 specifically binds CSR-1. EFT-3 was present in both FBF-1 and control IPs (Fig. 1a), so we performed further assays to confirm specificity of binding (see below). Our co-IP experiments suggested that CSR-1 should be expressed in the same germline region as FBF. FBF is enriched in the germ cell cytoplasm at the distal end of the gonad25, and we found CSR-1 in the same location (Fig. 1b), in addition to its known location in the proximal germline20. Since FBF-1 and CSR-1 co-IP and are expressed in the same region, FBF-1 and CSR-1 likely interact in vivo.
PUF and Ago family members both repress translation4,10, and models have suggested that miRNA binding sites proximal to PUF mRNA target sites may indicate an interaction26. To ask if FBF and CSR-1 might function together, we examined expression of an FBF reporter mRNA after CSR-1 depletion. In the reporter mRNA, the 3′ UTR of gld-1, an FBF target mRNA25, was linked to an ORF encoding a GFP histone H2B fusion protein (GFP::H2B), which labels nuclei. We used RNAi to deplete csr-1 from animals transgenic for reporters with either the wild-type gld-1 3′ UTR (WT gld-1 3′ UTR) or a mutant gld-1 3′ UTR (ΔFBE gld-1 3′ UTR) without FBF binding elements (FBEs)27. Briefly, fourth-stage larvae were subjected to feeding RNAi, and the GFP::H2B reporter was scored one generation later in adult progeny. In control animals not depleted for CSR-1, GFP::H2B was absent from the distal-most germ cells with the WT gld-1 3′ UTR reporter (Fig. 1c, top panels), but GFP::H2B extended to the distal end in animals with the ΔFBE gld-1 3′ UTR reporter (Fig. 1c, bottom panels), as seen previously27. After CSR-1 depletion in animals carrying the WT gld-1 3′ UTR reporter, bright GFP::H2B expression was always shifted distally compared to controls (n = 31/31) (Fig. 1d, top panels) and was often in distal-most germ cells (n = 12/31). By contrast, csr-1 RNAi had little effect on expression of the ΔFBE gld-1 3′ UTR reporter (Fig. 1d, bottom panels). As a control, the same reporter assay was used after depleting proteins thought to form a complex with CSR-1 (ref. 24), but these depletions did not affect the WT gld-1 3′ UTR reporter (Figs. 1e, f). Because total loss of FBE-mediated repression always leads to bright GFP::H2B in the distal-most germ cells28, csr-1 depletion does not completely eliminate FBE-mediated repression but instead severely compromises that repression. We conclude that CSR-1 reduces expression of FBF reporter mRNA and that this effect requires FBEs.
To validate our findings with the gld-1 reporter mRNAs, we assayed two FBF target mRNAs, gld-1 (ref. 25) and cye-1 (ref. 29) mRNAs, in a csr-1 mutant. GLD-1 protein is normally expressed at a low but graded level in the mitotic zone (Supplementary Fig. 2a, top panel), while CYE-1 is more uniform in the mitotic zone (Supplementary Fig. 2a, middle panel). In a csr-1 deletion mutant, GLD-1 and CSR-1 both increased dramatically in the mitotic zone, though GLD-1 was still graded (Supplementary Fig. 2b, top and middle panels). For controls, we also analyzed GLD-1 and CYE-1 levels in mutants of drh-3, ego-1, and ekl-1, which are thought to function jointly with csr-1 (refs. 20,24,30). In these mutants, we observed more GLD-1 and CYE-1 than wild-type, but not to the high level typical of csr-1 mutants (Supplementary Figs. 2c–e). Therefore, wild-type CSR-1 lowers expression of at least two FBF target mRNAs.
These experiments suggest that FBF functions with CSR-1 to repress target mRNAs. Yet, the documented germline defects in fbf and csr-1 mutants differ in many respects. The fbf-1 fbf-2 double mutant germlines are defective for germline self-renewal and hence have no mitotic zone25 while csr-1 is defective for chromosome segregation and embryonic viability19. We examined csr-1 deletion mutants for mitotic zone defects and found the csr-1 mitotic zone to be present but smaller than normal (Supplementary Fig. 2b). The reduced csr-1 mitotic zone is consistent with a functional overlap with FBF, but its presence demonstrates that CSR-1 is not required for all FBF functions. We suggest that CSR-1 and FBF can work together to repress mRNA expression via FBEs, but that they can also work independently. The independent functions likely involve interactions with other paralogs (e.g. FBF with other Agos, CSR-1 with other PUFs) as well as interactions with other complexes (e.g. FBF with the Ccr4/Not deadenylase and CSR-1 with RISC).
To directly assay FBF-1 interactions with CSR-1, we performed biochemical analyses with purified recombinant proteins. Initially, we explored the interaction, focusing on conserved domains – the PUF repeats of FBF-1 (hereafter called FBF-1) and the MID-PIWI or PAZ domains of CSR-1. GST-FBF-1 (a.a. 121–612) was co-expressed with either His6-MID-PIWI (a.a. 557–993) or His6-PAZ (a.a. 378–505) in the pETDuet system, which allows simultaneous expression of multiple proteins. Affinity selection for GST-FBF-1 enriched His6-MID-PIWI (Fig. 2a, lane 1) but not His6-PAZ (Fig. 2b, lane 1); the reciprocal selection for His6-MID-PIWI enriched GST-FBF-1 (Fig. 2a, lane 2), but selection for His6-PAZ did not enrich GST-FBF-1 (Fig. 2b, lane 2). Therefore, purified FBF-1 interacts specifically with the CSR-1 MID-PIWI domains but not with its PAZ domain. Our next experiments employed the CSR-1 MID-PIWI region, referred to as CSR-1 for brevity.
This work identifies EFT-3 as a potential FBF-1 interactor (Supplementary Table 1), and work by others identified human eEF1A as a potential AGO1 and AGO2 interactor31, although the interaction was not confirmed for either nematode or human components. We hypothesized that the FBF-1/CSR-1 heterodimer might form a ternary complex with the translation elongation factor EFT-3. To explore that idea, we purified FBF-1 alone, the FBF-1/CSR-1 heterodimer and EFT-3 alone (Fig. 2c, lanes 1–4); attempts to purify CSR-1 by itself were unsuccessful. FBF-1 alone did not bind EFT-3 (Fig. 2c, lane 5), but the FBF-1/CSR-1 heterodimer did bind EFT-3 (Fig. 2c, lane 7), suggesting existence of a ternary complex. This ternary complex was observed in the absence of GTP (Fig. 2c, lane 7), but FBF-1/CSR-1 also bound EFT-3 when GTP was included (Supplementary Fig. 3a). We confirmed the identity of CSR-1 and EFT-3 proteins in these pull-down assays by western blotting (Supplementary Fig. 3b). We examined FBF-1 for conserved residues that are not involved in RNA-binding and found a broadly-conserved Phe residue (Phe344) (Supplementary Fig. 3c). We hypothesized that mutant FBF-1(F344R) might be defective for ternary complex formation. Although FBF-1(F344R) was able to bind CSR-1 (Supplementary Fig. 3d), the FBF-1(F344R)/CSR-1 heterodimer failed to bind EFT-3 (Fig. 2c, lane 8). We propose that FBF-1/CSR-1/EFT-3 exists as a complex.
How might the complex affect translation? eEF1A family members possess GTPase activity, which is required during translation elongation to release aminoacyl-tRNAs upon delivery to the ribosome. Purified eEF1A has intrinsic GTPase activity, which is stimulated by ribosomes32. We hypothesized that the PUF/Ago complex might inhibit eEF1A activity since PUF and Ago proteins can inhibit translation4,9–11. To test this idea, we first confirmed that wild-type EFT-3 possesses GTPase activity compared to a catalytically-dead mutant EFT-3(H95L)33 (Fig. 2d; Supplementary Fig. 3e). We then assayed FBF-1 alone and the FBF-1/CSR-1 heterodimer for an effect on EFT-3 GTPase activity. FBF-1 alone did not reduce GTPase activity, but the FBF-1/CSR-1 heterodimer reduced GTPase activity to a level similar to that of catalytically-dead EFT-3(H95L) (Fig. 2d). Importantly, the mutant FBF-1(F344R)/CSR-1 heterodimer, which did not form a complex with EFT-3, also did not inhibit EFT-3 GTPase activity (Fig. 2d). Inhibition of EFT-3 GTPase activity in the FBF-1/CSR-1/EFT-3 complex suggests that the complex may repress translational elongation.
We predicted that a PUM2/AGO/eEF1A complex might also form in humans. Initially, we tested whether PUM2 binds AGO family members in vivo. HEK293T cells were transiently transfected with constructs encoding the Flag-tagged PUF repeats of human PUM2 (PUM2-3xFlag) and Flag-HA-tagged, full-length human AGOs1-3 (ref. 31). Using anti-HA antibodies, AGOs1-3 were immunoprecipitated from formaldehyde-crosslinked cells under denaturing conditions with RNase to isolate endogenous complexes34. Anti-Flag western blots revealed that PUM2 and AGOs1-3 co-IP (Supplementary Fig. 4a). We confirmed that PUM2 co-IPs with AGOs1-3 without crosslinking (Supplementary Fig. 4b). To test whether PUM2 is critical for the association of Ago and eEF1A, we performed the same transfection experiments as above in the presence or absence of PUM2. When PUM2 was co-expressed, AGO1-3 IPs pulled down dramatically more eEF1A than when PUM2 was omitted, arguing that PUM2/AGO binds eEF1A in vivo (Fig. 3a). We conclude that the PUM2/AGO/eEF1A complex is conserved.
We turned to rabbit reticulocyte lysate to characterize PUM2/Ago/eEF1A effects on translation. For these experiments, we first identified PUM2 mutants that abolish complex formation or RNA binding. Each PUM2 variant tested was comprised of the PUF repeats of human PUM2 (a.a. 706–1062) and was generated as a GST-PUM2 fusion protein. PUM2 refers to the wild-type version; PUM2(F868R) alters the conserved Phe corresponding to FBF-1(F344); PUM2(T752E) changes a conserved Thr residue (Supplementary Figs. 5a–c); and PUM2(H852A) was predicted to abrogate RNA binding35. Of these four proteins (Supplementary Fig. 5d), only PUM2(H852A) failed to bind an RNA carrying PUF-binding elements (PBEs) (Fig. 3b). When assayed for complex formation, wild-type PUM2 and PUM2(H852A) co-precipitated Ago and eEF1A from rabbit reticulocyte lysate (Fig. 3c, lanes 3 and 6). PUM2(F868R) bound Ago, but not eEF1A (Fig. 3c, lane 4), and PUM2(T752E) failed to bind either Ago or eEF1A (Fig. 3c, lane 5). In summary, PUM2(T752E) is Ago-binding defective (ABD); PUM2(F868R) is eEF1A-binding defective (EBD); and PUM2(H852A) is RNA-binding defective (RBD) but binds both Ago and eEF1A (Fig. 3d). The PUM2/Ago heterodimer recruits eEF1A, and PUM2 RNA-binding activity is not required for complex formation.
To assay effects of the PUM2/Ago/eEF1A complex on translation, we used reporter mRNAs in reticulocyte lysate. We first quantitated Ago levels in the lysate (Supplementary Fig. 6a) and added an equivalent amount of PUM2. To ensure that reticulocyte lysate was not limiting for translation, we titrated mRNA encoding firefly luciferase with 3xPBEs in its 3′ UTR [PBEs bind human PUM1 and PUM2 (refs. 36,37)] against constant control Renilla luciferase reporter lacking PBEs (Supplementary Fig. 6b). PUM2 did not destabilize reporter mRNAs; in fact, PUM2 stabilized them (Supplementary Fig. 6c). Therefore, the ratio of firefly to Renilla luciferase enzyme activities monitors PBE-dependent translation. We first assayed nonadenylated mRNA (Fig. 3e) to exclude from our analysis PUF-mediated deadenylation effects5. Wild-type PUM2 inhibited mRNA translation, but the three mutant PUM2 proteins failed to repress reporter expression (Fig. 3e). Most importantly, the ABD and EBD mutants, which disrupt PUF/Ago/eEF1A complex formation (Fig. 3b), did not repress the reporter.
Most natural mRNAs are polyadenylated, so we also assayed polyadenylated reporter mRNAs (Fig. 3f). Similar to the effects observed on the nonadenylated reporter mRNA, wild-type PUM2 repressed translation of polyadenylated mRNA while PUM2(RBD) did not. Notably, the PUM2 ABD and EBD mutants remained capable of limited repression. One possibility might have been that the ABD and EBD mutants had this minor effect by promoting mRNA deadenylation. However, mRNA adenylation was unaffected in our assays (Supplementary Fig. 6d). An open question is how PUM2(ABD and EBD) repress the polyadenylated reporter in the absence of both deadenylation and PUM2/Ago/eEF1A complex formation. Indeed, these mutants suggest the existence of yet another mechanism of PUF repression. Since PUF proteins associate with Nanos38, we considered the possibility that the region of PUM2 important for its Nanos association might be important for PUM2 repression. However, PUM2(G987D) repressed both nonadenylated and polyadenylated reporter mRNAs as well as wild-type PUM2 (Supplementary Fig. 7). Together, experiments using nonadenylated and polyadenylated mRNA reporters argue that the PUF/Ago/eEF1A complex represses translation and that it does so independent of mRNA deadenylation.
We next investigated the mechanism of PUM2 translation inhibition, again in rabbit reticulocyte lysates. Initially, we performed a standard assay for inhibition of translation initiation: we fractionated polyribosomes to ask if PUM2 inhibits the association of its target mRNA with ribosomes. In vitro translation reactions were performed as above with firefly mRNA (3xPBEs) and Renilla mRNA (Control) in the presence of PUM2 or PUM2(RBD). Reactions were quenched with cycloheximide (maintains ribosomes on mRNA) or puromycin (removes ribosomes from mRNA) and separated over a sucrose gradient. After centrifugation, polyribosome profiles were indistinguishable when PUM2 or PUM2(RBD) was added, although puromycin did disrupt polyribosomes (Supplementary Figs. 8a, b). Moreover, by Northern blots, the 3xPBE-containing mRNA associated with polyribosomes to the same level as the control mRNA if either PUM2 or PUM2(RBD) was added (Supplementary Fig. 8c). Since the 3xPBE-containing mRNA migrated in lighter fractions after puromycin addition, it is bound to ribosomes (Supplementary Fig. 8d). Although these data contrast observations that miRNA-mediated translation repression occurs at initiation in rabbit reticulocyte lysate39, we note that our experiments query PUF/Ago/eEF1A, not miRNAs. Therefore, ribosomal association with PUM2-bound mRNA appears unaffected by this assay, leading us to ask further questions about how translation is inhibited.
We next varied the translation assays to ask if PUM2-mediated repression occurs after translation initiation. Reporter RNAs plus purified recombinant PUM2 protein were incubated in lysate for 5 minutes to begin translation; then excess mRNA cap analog was added to quench additional ribosomal loading, and thus to block translation initiation. Production of firefly (subject to PUM2 regulation) and Renilla luciferase was monitored in the presence of either PUM2 or PUM2(RBD) over time. In the absence of cap analog, reporters produced luciferase linearly for about 45 minutes; however, in the presence of cap analog, luciferase production stopped much earlier. The control Renilla luciferase RNA plateaued after only ~13 minutes in the presence of either PUM2 or PUM2(RBD) (Fig. 4a). This early plateau indicates that the excess cap analog had abolished de novo 40S ribosomal subunit loading. With the RNA-binding defective PUM2 variant, expression from the firefly luciferase mRNA plateaued after ~15 minutes, a bit after the Renilla control, presumably because its protein product is larger. However, wild-type PUM2 had a dramatic effect on firefly luciferase expression, decreasing sharply after only 8 minutes (Fig. 4b, left panel), an effect that can account for the difference in synthesized protein observed in Figures 3e, f. Wild-type PUM2 also modestly, but reproducibly, delayed the initial production of firefly luciferase relative to PUM2(RBD), an effect that likely reflects slower translation elongation (Fig. 4b, right panel; summarized in Fig. 4c). That slowed translation elongation is consistent with inhibited eEF1A GTPase activity in the PUM2/Ago/eEF1A complex, but the effect was not large enough to account for the four-fold reduction in synthesized protein seen in Figures 3e, f. We conclude that PUM2 slows translation elongation and blocks protein production from its target mRNA.
We considered possible mechanisms by which PUM2 might inhibit protein production: it might attenuate translation elongation, consistent with the ribosome drop-off model proposed for miRNA-mediated translation repression15; it might inhibit translation termination; or it might promote destruction of nascent polypeptides during translation, as observed for miRNA-mediated repression40. The first two mechanisms make predictions about effects on ribosome position. If ribosomes drop off the mRNA during elongation, ribosomal density should be higher at the 5′ end of the ORF and taper off towards the 3′ end. If translation termination is inhibited, ribosomes should accumulate at the 3′ end of the ORF. High-resolution ribosomal footprinting was previously described at a genomic level41, and we modified this approach for our single mRNA reporter (Fig. 5a). Briefly, in vitro translation reactions were performed with radiolabeled firefly luciferase mRNA (with 3xPBEs). Reactions contained PUM2, PUM2(ABD), PUM2(EBD) or PUM2(RBD). Translation reactions were quenched with cycloheximide and treated with RNase One, then fractionated over a sucrose gradient to isolate ribosome-bound RNA fragments. These RNA fragments were extracted from the monoribosome peak and hybridized to a spotted array containing overlapping DNA oligonucleotides complementary to the firefly luciferase mRNA (Supplementary Fig. 9). RNA signals from monoribosome fractions were normalized to signal from unfractionated samples to control for hybridization efficiency, uridine content and RNase sensitivity and then plotted across the mRNA (Fig. 5b). Strikingly, in the presence of PUM2, a large peak occurs ~100–140 nts into the ORF, and then ribosomal footprints are sharply reduced across the remainder of the ORF. The peak coincides with the position at which nascent polypeptide should emerge from the ribosomal exit tunnel (~30–40 amino acids42). Importantly, the ribosomal footprints observed in samples with PUM2(ABD and EBD) were similar to that with PUM2(RBD). Therefore, the ribosomal footprinting profile is only altered when PUM2 can form a ternary complex on its target mRNA. To ensure that the observed ribosomal profile was independent of nucleotide sequence, we repeated our ribosomal footprinting experiment with firefly luciferase mRNA lacking ~500 nts after the initiation codon (including the sequence where ribosomes accumulated above). Again, we observed decreased ribosomal density after the first ~140 nts into the ORF only when wild-type PUM2 was added (Fig. 5c). We conclude that PUM2/Ago/eEF1A complex attenuates translation elongation, causing ribosomes to accumulate within the ORF.
Our data support three main conclusions that advance our understanding of how PUF and Ago proteins regulate translation. First, PUF and Ago proteins can form a conserved complex containing the core translation elongation factor, eEF1A. Second, the nematode complex inhibits eEF1A GTPase activity, and the human complex represses translation. Third, the complex blocks elongation at a site ~100–140 nts into the open reading frame, where the nascent peptide is expected to emerge from the ribosomal exit tunnel. Below we discuss the implications of these findings and propose a model to describe our observations: that PUF/Ago inhibits eEF1A GTPase activity and blocks translation elongation.
Studies prior to this work focused on either PUF or Ago proteins4,9–11. One hint that the two might work together was a bioinformatics analysis of human 3′ UTRs, revealing a nonrandom proximity of PUF protein binding elements and miRNA binding sites26. Here, we demonstrate not only that PUF and Ago proteins interact, but also that the PUF/Ago heterodimer associates with eEF1A and inhibits its GTPase activity. Our focus in nematodes was the FBF-1 PUF protein and the CSR-1 Ago protein, but we suspect that other PUF/Ago combinations may occur, as the nematode genome encodes 11 PUFs and 27 Agos. FBF-1 and CSR-1 are divergent members of their respective families, yet both proteins possess signature domains. Moreover, CSR-1 has slicing activity23 despite its non-canonical function in chromosome segregation19. One possibility might have been that the FBF-1/CSR-1 association was atypical. However in mammals, human PUM2 can associate with three AGOs (AGO1, AGO2 and AGO3), suggesting conservation. We suggest that multiple PUF/Ago combinations exist both in nematodes and mammals. A question for the future is whether individual PUFs and Agos behave differently with respect to their participation in the PUF/Ago/eEF1A complex.
Small RNAs cannot be essential for formation of the PUF/Ago/eEF1A complex. The MID-PIWI domain of Ago is sufficient to form the complex, and together with its PUF partner, the MID-PIWI domain inhibits eEF1A GTPase activity. Yet the MID-PIWI domain cannot bind small RNAs on its own. By contrast, the PAZ domain of Ago is not required to assemble the complex (Fig. 2c), but this PAZ domain drives small RNA loading43. Nonetheless, PUF/Ago/eEF1A complexes formed in vivo contain full-length Agos and likely associate with small RNAs. An attractive idea is that the PUF/Ago/eEF1A complex coordinates PUF and miRNA regulation.
The interaction between the PUF/Ago heterodimer and eEF1A complex is reminiscent of the original identification of Ago in a complex with eIF2 (ref. 44), which is structurally similar to eEF1A and also possesses GTPase activity. Moreover, two Drosophila RNA-binding proteins, dFXR and VIG, coordinate with Ago2 in the context of miRISC to repress target mRNAs45. An exciting possibility is that Ago proteins can work with other RNA-binding proteins and other translation factors to modulate multiple steps of protein synthesis.
Figure 5d presents a model for how the PUF/Ago/eEF1A complex may control translation elongation. Central to this model is its anchor via the PUF/Ago heterodimer to regulatory elements in the 3′ UTR and its association with an inhibited core translation elongation factor, eEF1A. Also central to this model is a block within the ORF at a position corresponding roughly to where the nascent polypeptide emerges from the ribosomal exit tunnel. Our kinetic analysis of protein production from a PUM2-repressed reporter mRNA (Fig. 4) suggested two phases of translational inhibition. An initial modest inhibition was inferred from a slower elongation rate, consistent with inhibition of eEF1A GTPase activity; a subsequent full repression was inferred from a complete block in protein synthesis seen later. Full repression may require a conformational change in the PUF/Ago/eEF1A complex, its modification, or recruitment of additional factors. If ribosomes do indeed accumulate where the nascent polypeptide leaves the ribosomal exit tunnel, an additional factor that recognizes the nascent polypeptide seems likely.
The model in Figure 5d does not include any small RNA associated with Ago, because our work focused on Ago, PUF and eEF1A proteins, not small RNAs. For example, a miRNA binding site was not engineered into the reporter 3′ UTRs, and miRNAs were not added to the reactions. One intriguing possibility is that the PUF/Ago/eEF1A mechanism reveals a role for Ago that does not rely on small RNAs. Perhaps more likely is the idea that small RNAs influence recruitment, either in an essential or facilitating manner. Addressing the role of small RNAs in this mechanism is an obvious next step for future analyses.
Repression at the level of translation initiation is emerging as a favored mechanism of translational control by miRNAs9–11. We provide two lines of evidence that the PUF/Ago/eEF1A complex does not have a major effect on translation initiation. First, polyribosomal profiles were the same for a PUM2-repressed mRNA and a control mRNA (Supplementary Fig. 8). Second, the same initial slope was observed in our kinetic analysis of protein production by PUM2-repressed and control mRNAs (Fig. 4). In addition, PUM2 repression occurred in the presence of excess mRNA cap analog, suggesting that its repression is exerted after loading 40S ribosomal subunits. Although we cannot exclude the possibility of a minor effect on translation initiation, our results strongly point to translational elongation as the major level at which the PUF/Ago/eEF1A complex exerts its repressive effects.
How does the proposed PUF/Ago/eEF1A mechanism compare to established mechanisms of regulation at the level of translation elongation? Perhaps the best-known mechanism is that adopted by the signal recognition particle (SRP) during synthesis of membrane proteins46, where translation elongation is inhibited47 until SRP docks with the endoplasmic reticulum46. The PUF/Ago/eEF1A mechanism resembles this case in that ribosomes accumulate at a similar position within the ORF. However, no role for 3′ UTR binding proteins is known for the SRP block. Recently discovered is the hnRNP E1 association with eEF1A to inhibit translation elongation; however, hnRNP E1 does not inhibit eEF1A GTPase activity but instead inhibits eEF1A dissociation from the ribosome48. Another case is miRISC, which is thought to promote ribosomal drop-off from a targeted mRNA but not at a given position within the ORF15. Therefore no established mechanism fits the PUF/Ago/eEF1A model completely.
The PUF/Ago/eEF1A mechanism controlling translation elongation (Fig. 5d) is not the only mechanism used by PUF and Ago proteins, and perhaps not even the primary mechanism. Its relative importance to other well-established mechanisms (e.g. deadenylation and instability, see Introduction) remains unknown. Why employ multiple mechanisms? We do not know but suggest three ideas. One is security. If the PUF protein or Ago fails to silence a target mRNA with one mechanism, the regulator may invoke a different mechanism to repress the escaped mRNA. A second idea is reversibility. Destruction is a dead end for an mRNA, but blocked elongation might ensure robust activation at a later step. A third is subcellular compartmentalization. As an mRNA emerges from the nucleus, it meets one subcellular environment, but upon localization, for example to stress granules, the mRNA enters a different environment. Translational repression in (or localization to) distinct subcellular locales may differentially employ one mechanism over another. Key issues for the future include unraveling the biological functions of the various PUF and Ago mechanisms, their relationships and their regulation.
We thank H. Tabara (Kyoto University) for providing CSR-1 and DRH-3 antibodies and E. Kipreos (University of Georgia) for CYE-1 antibody. We thank Kimble and Wickens lab members for discussion; we also thank Elsebet Lund and Scott Kennedy for critical reading of the manuscript and Anne Helsley-Marchbanks and Laura Vanderploeg for help with the manuscript and figure preparation. Mass spectrometry was performed with support from the Human Proteomics Program at the University of Wisconsin-Madison.
Note: Supplementary information is available on the Nature Structural & Molecular Biology website.
AUTHOR CONTRIBUTIONSK.F. performed the experiments with the exception of phylogenetic analysis (Z.C.), luciferase reporter mRNA production (A.C.) and csr-1 mutant generation (P.K.-C.). K.F., M.W. and J.K. prepared the manuscript. K.F. is supported by PF-10-127-01-DDC from the American Cancer Society, Z.C. by National Institutes of Health postdoctoral fellowship F32 GM095169, A.C. by N.I.H. training grant T32 GM07215 and an Advanced Opportunity Fellowship from UW-Madison, M.W. by N.I.H. grants GM031892 and GM050942 and J.K. by N.I.H. grant GM069454. J.K. is an Investigator of the Howard Hughes Medical Institute.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Stage - 2003 Rock Around the Church
A 'Christians in Entertainment' show, with Cannon and Ball, Syd Little, Diane Regan and compered by CiE organiser Chris Gidney.
Christians in Entertainment has produced a souvenir brochure for their shows. Called 'The Laughter Makers' it includes biographies of a number of Christian entertainers.
As this brochure is, at the time of writing, still commercially available, none of it is duplicated on this site. The page showing Cannon and Ball is pictured below.
Known Dates
|Monday 6th Oct
|North Bridge Leisure Centre, Halifax
|Friday 10th Oct
|Fuller Baptist Church, Kettering
There were undoubtedly other dates!
|Sunday 23rd May
|Bracknell Family Church, Bracknell
|Monday 24th May
|Princess Royal Theatre, Briton Ferry
|Tuesday 25th May
|Holy Trinity Church, Attleborough, Nuneaton
|Wednesday 26th May
Thursday 27th May
|Tytherington Family Worship Church, Macclesfield
|Friday 28th May
|New Life Christian Fellowship, Lincoln
These photos were taken at the performance in Halifax, 6th October 2003.
|Compere Chris Gidney
|Singer Diane Regan
|Guitarist/Singer Syd Little
|Tommy Cannon
|Bobby Ball
This backstage cast photo is from Christians in Entertainment

subset: commoncrawl


text: Special Reviews Genome Sequence of the Nematode C. elegans: A Platform for Investigating Biology The C. elegans Sequencing Consortium* See allHide authors and affiliations Science 11 Dec 1998:Vol. 282, Issue 5396, pp. 2012-2018DOI: 10.1126/science.282.5396.2012 Article Figures & Data Info & Metrics eLetters PDF This article has corrections. Please see:Corrections and Clarifications - January 01, 1999 CORRECTIONS AND CLARIFICATIONS - September 03, 1999 Log in to view full text Via AAAS ID This article is available to AAAS members. If you are a AAAS Member use your via AAAS ID and password to log in. Not a member? Click here to join. Join/Subscribe Purchase Article Activate Member Account Renew Subscription Recommend a subscription to your library Help for librarians Order a single issue


text:
SuperGuarantee: SuperMedia ServiceMagic wannabe that isn't
01 Mar 2010
Supermedia, the former Idearc, just released a multi-faceted announcement about its one-year old SuperGuarantee, its new SuperGuarantee Autos, and the SuperPages mobile application. We'll soon be talking to SuperMedia about the new Autos product, but took a first look at SuperGuarantee in the meantime. The world of online directo


text:
The college was our family. we had a nice and fun ti, e in college. It was all mixed feelings. I can only say that Days cannot be better than those day.
I took an extrance examination of CEE, however I could not able to join any medical college so I decided to choose any stream something similar to which I want i, e none other than Biotechnology. Also I searched in google and found that Bangalore is the ideal location in order to conduct the course. So I selected Bangalore and also I wanted to join a College where it should be quite pocket friendly for all my expenses. Thus I selected St. George College and it is the best decision of my Life. Wh...
No Comments To Show
2017-06-23 15:07:41
Bangalore University was established in July 1964 in the garden city of Bangalore with a vision of promoting excellence in education. University offers various programs such as Undergraduate, postgraduate, Ph.D. and integrated courses. It invites applications for admissions to various PG courses
2017-06-13 17:28:47
Bangalore University, Bangalore invites applications for admission to 5 Years Bachelor of Arts Bachelor of Law (B.A LL.B) Program in the University Law College, Jnana Bharathi Campus, Bangalore University for the academic year 2017-18. Admission will be done on the basis of meri
2017-03-07 14:52:39
Bangalore University (BU) Conducts the semester examination in the months of May/June & November/December every year. The declaration of the result depends on the aggrega
2016-07-14 17:10:19
B.A. L.L.B Notification - Bangalore University
2016-07-14 17:10:07
Download Prospectus - Bangalore University
2016-07-14 17:09:50
Official Website of Bangalore University
2015-06-02 14:24:45
Bangalore University invites the application for the 1st Semester of Five Year B.A. LL.B. Degree for the academic year 2015-16 in the


text:
Explore Matt Bomer, White Collar and more!
Explore related topics
Matt Bomer is SO pretty!
oh Matt Bomer how I love you." The White Collar star is basically the closest thing to a real-life Disney prince ever. Why settle for mediocre when Bomer's basically the real thing? (Seriously, those eyes. And face. And hair. And smile. And abs.
Matt Bomer aka my Christian Grey
Matt Bomer freakin beautiful - My favorite Christian Grey! The only Christian Grey!
MATT BOMER not Christian grey! He might be playing Christian Grey in "Fifty Shades of Grey." Matt Bomer is the star of the USA Network television series White Collar and plays ken in the movie Magic Mike.
Matt Bomer as Neal Caffrey - White Collar and Christian Grey - 50 Shades of Grey
Matt Bomer-tv-show"White collar". never get tired of looking at this face.What a doll can we clone him to get a heterosexual likeness to share?
Matt Bomer,,,Marry me already?
Boardwalk Babe from Matt Bomer's Hottest Pics The Golden Globe winner keeps a pensive look on his face—those are some serious abs, after all—while on vacation in Maui.


text:
- Design, Production, Staffing AGENC Experiential + Digital Marketing (formerly Caravents)
- DJ DJ Rashida
- Flowers R. Jack Balthazar
- Lighting Lighten Up Inc.
- PR BNC (Bragman Nyman Cafarelli)—Los Angeles
- Press Risers, Ropes, Stanchions Classic Party Rentals
- Security Meyerson Associates Inc.
- Venue Voyeur
Search our directory
LOS ANGELES On Wednesday, Us Weekly toasted its annual Hot Hollywood issue with a celebrity-filled event at West Hollywood's new Voyeur. After a stint in New York, the party came back to Los Angeles this year, and its latest incarnation stuck to its known formula: introducing guests to a mostly yet-unseen, buzzed-about new nightclub. This time, the dark, broody Voyeur set the stage for the program.
The mag's honorees in attendance included "Comeback of the Year" Kristen Cavallari, "Victor of the Year" Kris Allen, and "Glam God of the Year" Adam Lambert, and the cast of Glee, which Us named "Cast of the Year." Wenner Media's Keira Ford oversaw the event, tapping Caravents to produce it once again.
Dancers styled a la Eyes Wide Shut performed throughout the space, and leather curtains and velvet furnishings gave the party a burlesque feel. Caravents used decor in burgundy and crimson to inspire an autumnal mood—not to mention a bit of Hollywood drama—and a golden carpet sprawled beneath a 32-foot step-and-repeat. After celebrities walked the arrivals line, they headed down the carpet into a crimson-draped photo studio to have their mugs snapped for use in the following week's Hot Hollywood issue.
Next, they proceeded into the gifting suite, where True Religion hawked its wares and Vaseline showcased its new lotion line, Sheer Infusions; the brand also offered mini hand massages inside the party. Sponsor Skyy poured its cocktails, and drinks from sponsors Miller Genuine Draft and Snapple were also available at the bars. To integrate sponsor Yahoo, Caravents set up a logo photo booth inside the venue that allowed guests to email shots to their friends and phones, right from the party.
DJ Rashida spun for guests like Adam Lambert, Leona Lewis, and Whitney Port (who organizers worked to keep away from Stephanie Pratt—as such details are important in Hollywood). Overhead, a mesh canopy supported a writhing, scantily clad live model.


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How Do You Avoid A Motorcycle Accident?
Uploaded by Lucia, found in Blogs
Besides taking basic safety precautions like following the speed limit and wearing a helmet, there are ways motorcyclists - as well as other motorists - can help decrease the number of motorcycle-related wrecks on our roadways. Here's a few tips to consider:
Never drink and drive
While you cannot control what others do, including those in cars, you can make sure you are coherent and obeying the law by driving sober.
Stay aware of your surroundings
In addition to forming a plan before leaving, make sure to stay aware of your surroundings when traveling. Is there anyone traveling in front of or behind you? What is the terrain; can you pull over if necessary? Where was the last gas station or rest area you passed? Do not drink and drive. A great percentage of motorcycle-related accidents are alcohol related.
Be proactive and plan
Know where you're going before you leave. Even if you're only going on a short drive, have a plan. Will you be stopping for lunch? If so, where and about what time? If you're going alone let someone know where you're going and what time they can expect you to return. If you're going on a longer trip, map it out. Have a detailed plan. How will traffic be on one route vs. another route? Is there road construction under way anywhere on your route? What is the weather forecast? The more you know before you leave, the better chance you'll have of getting to your destination safely.
Take a class
Local groups and some DMVs offer motorcycle safety classes and programs. If you don't have time for a full-on class, go online and check out some websites. There are many that offer tips and advice that, if followed, can help keep everyone traveling our roadways safe.
Educate your passenger
If a passenger is traveling with you tell them what to expect and what you expect before getting on the road. This especially applies to first-time or novice riders. Passengers who plan to ride often could also take a class or go online to learn about responsible and safe riding.


text:
Coalfields Highway truck plan a 'safety issue'
The Collie Shire president has raised concerns over a plan to move hundreds of extra trucks per week through a prominent south-west highway.
An agribusiness is believed to be planning to transport wheat to the Port of Bunbury, on the Coalfields Highway, via Collie.
Wayne Sanford says he is concerned about the condition of the highway and the extra trucks movements would put the community at risk.
"It just needs to have a level of consultation around what the impacts might be for not only Collie but other residents that live along the Coalfields Highway and I'm certainly concerned from a safety perspective and I'm certainly concerned about the ability of the Coalfields Highway to be upgraded," he said.
Mr Sanford says extra trucks would seriously damage the highway, which is already under pressure.
"The whole of the Coalfields Highway is under pressure certainly from a safety perspective both on the road and actually trying to cross the Coalfields Highway in the middle of the CBD of Collie, it's certainly a safety issue for the community," he said.
Topics: activism-and-lobbying, agribusiness, wheat, local-government, collie-6225, bunbury-6230


text:
map of the Bahamas
map of Caribbean
Copyright © All World
Vacation Station
Wedding Tips For Your Honeymoon by Rafi Michael
Bahamas A slice of commonwealth Britain in paradise by
John Gibb
life with the Grand Bahamas Island by Suzane Gray
to Bahamas Home Page
Hotels - Featured Bahamas Vacation Rentals
Bahamas Wedding Tips For
Your Honeymoon by Rafi Michael
The most exciting way to
begin your married life is to go to the Bahamas for your honeymoon. You
can flavor pleasure as you slurp icy drinks on a spongy sand beach. After
the chaotic activity of an exhilarating wedding you will search out a new
meaning of leisure on a Bahamian holiday. There's no better way to begin
the new chapter of your life than enjoying the fun, sun, and each other.
Though planning your wedding
you must give the same importance to the planning of your honeymoon also.
If you don't, you might have to resort to the traditional holiday and spend
more than your budget permits. After the hectic events of the wedding you
are probable to feel bushed, annoyed and tense. A honeymoon must be a relaxing
holiday for the two of you when you can leave all the problems behind and
focus on your new relationship.
Floating in Crystal Waters in Front of Seaplane, Bahamas
at AllPosters.com
Although planning your honeymoon,
consider your budget to start with. If you have already lengthened your
wedding budget, you will have to save individually to make your dream honeymoon
feasible. For that you can go to any one of the websites where you can
register towards a trip instead of the typical household items. This way
you will not only carry out your goal but also make sure you receive gifts
that you and your partner will use and enjoy. If this doesn't work, you
may want to inquire for financial gifts or airline mile donations to make
your Bahamas honeymoon happen.
Once you make up your mind
on going to the Bahamas for your honeymoon, you will find various destinations,
resorts, hotels, and vacation options to select from. You can search these
options on the World Wide Web or talk to your local travel agent to find
the best vacation for you and is within your budget. Planning your wedding
and honeymoon together can decrease the pressures and stresses of wedding
planning to a great extent.
It's a nice idea to go in
for an all comprehensive resort for your Bahamian holiday that includes
food and activity costs with the room fees. Most of these resorts are very
logically priced and give massive opportunities to enjoy all aspects of
the fun and sun of the islands. They are perfect for newly married couples
because they allow you the freedom from cost constraints.
One more well-liked way to
enjoy the Bahamas on your honeymoon on a tight budget is to take a cruise.
Many lavishness cruise companies offer various destinations in the Bahamas
and other islands in the Caribbean. Gone are the days when cruises were
considered to be just for old people looking for sun! These days you see
more young people on cruises, particularly, honeymooners and animated couples
that enjoy exhilarating nightlife and lovely cuisine.
The Bahamas are also the
favorite location for destination weddings. The amazing beauty of these
destinations provides a romantic aura to the ceremony. If you plan a destination
wedding you can comprise your loved ones in your holiday plans. And, if
there are children, you have more reasons to think of the Bahamas as kids
have great fun on the white sand in warm weather.
All in all, the Bahamas is
a great option for your honeymoon destination. It's the best way to begin
your new life together. This fantastic island will open out to you as you
relax on the beach, or indulge in the copious activities.
Hotels - Featured Bahamas Vacation Rentals
to Bahamas Home Page
Do you own quality vacation
accommodation in the Bahamas?
Why not Join
Quick Links:
- Antigua & Barbuda - Aruba
- Bahamas - Barbados
- Bermuda - Bonaire
- British Virgin Islands - Cayman
Islands - Cuba - Curacao
- Dominica - Dominican
Republic - Guadeloupe - Grenada
- Jamaica - Puerto
Rico - St Barts - St
Croix - St John - St
Kitts & Nevis - St Lucia - St
Martin/St Maarten - St Thomas - St
Vincent & the Grenadines - Tortola
- Trinidad & Tobago - Turks
& Caicos - Virgin Gorda - US
Virgin Islands


text:
Apartment Hebnes Finnvik II
Located in Hebnes, this apartment is 44 km from Haugesund. The property features views of the sea is 13 km from Sand. Free private parking is available on site.
The kitchen is fitted with a dishwasher, a microwave and a fridge, as well as a coffee machine. A TV is available. Apartment Hebnes Finnvik II features free WiFi throughout the property.
The nearest airport is Haugesund , Karmøy Airport, 48 km from the property.


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Harington has the title role in the drama, which involves a Hollywood star's career being destroyed when people start making assumptions about his correspondence with an 11-year-old actor. Chastain is set to play the editor of a gossip rag. Portman and Hoult's roles, meanwhile, have yet to be revealed.
Dolan immediately followed his festival hit Mommy (2014) with It's Only The End Of The World. That one stars Léa Seydoux (Blue Is The Warmest Colour), Marion Cotillard (Rust And Bone) and Vincent Cassel (Black Swan) and is currently in post-production for a release sometime this year.
We will next see Portman and Hoult, meanwhile, during the same weekend in the spring. Portman has the long delayed Western Jane Got A Gun, which arrives on 20 May, while Hoult will reprise his role as Beast in Bryan Singer's X-Men: Days Of Future Past (2014) follow-up X-Men: Apocalypse on 19 May.


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Support Us
Support the Future of the arts at the Morean Arts Center
Over the past century, St. Petersburg's deep and unwavering commitment to arts education has helped propel artists to national recognition. We have become a community of self-expression. The Morean Arts Center strengthens the region's arts community from every possible angle. It nurtures local artists, brings national and international artists to our community, celebrates the creative process, and cultivates a new generation of creators and collectors. With your help we can continue this tradition as we train, cultivate and produce the aspiring artists of our community.
Each year the Morean curates more than 30 exhibitions of work in all media by a broad spectrum of artists, including those who are emerging, mid-career and established, as well as work by children. Exhibition & Masterwork sponsorships cover the period of July 1st to June 30th.
As a community-based visual arts organization that has been part of the St. Petersburg cultural landscape since 1917, the Morean Arts Center, Glass Studio and Hot Shop, Morean Center for Clay and Chihuly Collection connect people with art through daily programming, offered 363 days each year.
We provide studio art classes in all media for all ages and abilities; art-making demonstrations in ceramics and glassblowing; K-12 and youth art education programs; and contemporary art exhibitions with influential and local artists. In 2016, we welcomed more than 130,000 visitors to one or more of our 4 locations in St. Petersburg, Florida.
Your support helps guarantee that future generations will not only be inspired by the art created and exhibited at one of the Morean's four venues, but also has the ability to let that inspiration enable the artists of tomorrow.
We hope you share our profound belief in the need for St. Petersburg to have a vibrant and significant community art center which enriches our community and quality of life. Your gift can be the difference in a young artist being able to pursue their talents, help to fund upgrades to programming, or help a family of modest means offer their child a level of expression they may otherwise never know.Without donors like you, we would not be able to provide the exceptional programs that we do today. We urge you to consider supporting the Morean Arts Center. Simply click here.
Other great ways to give include:
Gifts of Securities
Stocks, bonds, and mutual funds that you have owned for more than one year and that have increased in value can be a substantial gift, with a low net cost to you. You receive a charitable deduction for the donation, which is based on the stock's fair-market value on the date of the gift.
In addition, you avoid the federal Capital Gains Tax that would otherwise be owed on a sale of the asset. If the stock is worth less than you paid for it, it might be advantageous for you to consider selling it and using the cash proceeds to make a charitable gift. The amount of the gift can be a tax deduction, and the loss created by the sale may be used as a deduction from other taxable income.
The simplest way to donate stock to is through your broker. Your broker can transfer your stock to the Morean Arts Center using the following information:
Account #304-33725
Contacts: Julie Fisher, 727-825-7726
Address Info: 100 Second Ave. S., Suite 800, St. Petersburg, Fl 33701
E-mail: firstname.lastname@example.com
Please contact Anna Kuhlman at 727-822-7872 ext. 2120 or firstname.lastname@example.com to tell us about the gift so we can accurately attribute it to you.
Corporate Giving
The Morean welcomes corporate partnerships in the support of its educational mission to offer the highest quality art education and experience to all our students. Corporate contributions can assist in meeting our operating costs, bringing quality exhibitions for student and public enrichment, attracting the best students through scholarship donations, or by supporting one of the Morean's signature events. Corporate involvement and support of the Morean offers a visible distinction that emphasizes a priority to arts and community enhancement. We encourage your company to play a vital role by partnering with the Morean Arts Center.
Matching Gifts
Many companies offer matching gift programs to encourage employees to contribute to charitable organizations. Some provide matching funds to support employee volunteer hours. Most of these programs match contributions dollar for dollar, and some will even double or triple the amount of your gift!
To learn how you can support the Morean Arts Center, please contact:
email@example.com OR 727.822.7872 ext.2120


text:
Brooks-Range Dial Stem Thermometer
Where to Buy
sponsored links
Help support this site by making your next gear purchase through one of the links above. Click a link, buy what you need, and the seller will contribute a portion of the purchase price to support Trailspace's independent gear reviews.
Retailers' Descriptions
Here's what other sites are saying:
Our Dial Stem Thermometers read degrees in Celsius only since it is the standard scale for international studies in snow science. It comes with an easy-to-read one-inch diameter dial and five-inch long stem. The stainless steel stem has adjustment for calibration. Includes a case with a clip.Specs:Scale: 1 deg increments from -40 to +70 deg CelsiusSize: 1 in. diameter x 5 in. long
- Second Ascent
Similar Products
Brooks-Range Digital Thermometer
Backcountry Access Digital Thermometer
Backcountry Access Snow Study Kit
Brooks-Range Snow Study Organizer Pro
Backcountry Access Analog Thermometer
Backcountry Access Ruler
Brooks-Range Pocket Scale SR-1
Backcountry Access Slope Meter
Brooks-Range Dial Stem Thermometer
previously retailed for:
$7.95 - $12.95
$7.95 - $12.95
The Brooks-Range Dial Stem Thermometer is not available from the stores we monitor. It was last seen January 27, 2017 at CampSaver.com.


text:
All home afternoon match days (3.00pm kick off)
12.00 – 2.30pm and immediately after the match - 7.45pm
All home evening match days (7.45 kick off)
5.30 - 7.30pm and immediately after the match - 11pm
12 Evening Home v FC Halifax Town
13 Meet the Chair and Vice Chair (ticket only - see info on left)
16 Home v Hartlepool United
K/O 1230. Club open from 11.00am
30 Home v AFC Fylde
4 Tranmere v Orient live on TV, Club opens 5.30pm and closes 30
mins after the end of the game.
21 Home v Macclesfield Town
24 Evening Home v Gateshead
18 Home v Dover Athletic
21 Evening Home v Chester
9 Home v Sutton United
26 K/O 3.00pm v Dagenham and Redbridge
30 Home v Bromley
Latest News .......
>> PLEASE NOTE: In light of the move of the Tranmere game for TV, the planned coach will now NOT
RUN. Instead, the game will be shown on TV in the Club.
>> Membership form for 2017/18 is available under the How to Join tab above.
An evening with
the Chair
the Vice Chair
We have arranged an evening with Nigel and Kent for Wednesday 13 September at 7.30pm. Once again, it will be a ticket-only event, with remaining tickets available from email@example.com.
This website was last updated on 8 September 2017
Leyton Orient Women
The fixture list for the Women's team has now been published. You can find full details by clicking here.


text:
More Youth Apprenticeships
The Wilders Guild is a team of fiber artists, farmers, and craftspeople. The Wilders Apprenticeship teaches skills that bridge the gap between surviving and thriving. Wilders help to restore the land that they use through ethical harvesting and sustainable cultivation.
During the Wilders Apprenticeship, we will learn a variety of skills that may include and are not limited to:
- Fiber Arts
- Transform raw wool into wearable items by learning to card, spin, knit and weave.
- Folk Craft
- Apprentices learn crafts of woodworking, natural building, animal husbandry, outdoor cooking, and even artisan cheese making. They train both to sustain the Village and help build it.
- Plant Identification and Restoration
- Learn to identify a wide variety of plants to eat, collect for your herbal apothecary and dye fabrics! Plus, our expert mentors teach the principles of ecological restoration, where we restore the land around us to its fullest realization of health and bounty.
- Outdoor Cooking
- Make an array of delicious foods that help feed the village, cooking over an open fire.
How it Works
One weekend a month, apprentices train in the field at our epic outdoor locations. Four of those weekends are Friday - Sunday overnight expeditions.
- Overnight Weekends:
- Check-in between 4:30 and 5 PM on Friday and check-out at 3:00 PM on Sunday.
- Single Field Days:
- Check-in between 8:45 and 9 AM on Saturday, and check-out at 4:30 PM that same day.
TuitionTuition is divided into monthly payments after a $75 deposit, or may be paid in full by August 15 for a 5% discount.
Grade 5 - 8 (Includes 4 Weekend Overnight Trips): Full program tuition is $1,950. Includes: 4 Friday - Sunday overnight weekends, 4 field day Saturdays, Community Celebration: Picnic Potluck, optional family holiday party, Trackers hoodie, access to our weekly family Skills Night, and Mora knife.


text:
Market Research Survey(2 Posts)
AHope200 Tue 07-Mar-17 12:26:41
AuntieStella Fri 17-Mar-17 07:48:36
Join the discussion
Registering is free, easy, and means you can join in the discussion, get discounts, win prizes and lots more.Register now
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text:
St. Olaf's Church, Jomala
|Jomala Church, Church of St. Olaf
Jomala kyrka, Sankt Olav kyrka (in Swedish)
Jomalan kirkko, Pyhän Olavin kirkko (in Finnish)
Church of St. Olaf
|Basic information
|Jomala, Aland Islands, Finland
|Evangelical Lutheran Church of Finland
|Architectural description
|Architectural style
|Medieval stone
|c. 1280
|red granite, limestone
The Church of St. Olaf is a medieval stone church in Jomala, Åland Islands. It belongs to the Evangelical Lutheran Church of Finland. Dating from about 1260 to 1280, it is possibly the oldest Christian church in Finland, and was extended in the 19th century.[1] It is constructed of local red granite and limestone. The church is dedicated to King Olaf II of Norway, patron saint of the Åland Islands and Jomala.
The coloured wall paintings from the 1280s on the Western gable represent scenes from the parable of the Prodigal Son. Considerations to expand the church began in the 17th century and a plan from 1809, while under Swedish rule, was not acted on. From 1844, under the Russian Empire, the extensions were made. In 1968, a new stained-glass window was donated to the church by the American artist Thure Bengts, whose ancestors were from Jomala.[2]
See also[edit]
|Wikimedia Commons has media related to Jomala Church.
Coordinates: 60°09′18″N 19°56′54″E / 60.15500°N 19.94833°E


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Pulau Tenggol Diving Package & Resort Package
Pulau Tenggol Diving Package
Pulau Tenggol is a perfect island for divers, and you can find the Pulau Tenggol Diving Package here. Tenggol Island is an additional beautiful diving getaway island merely 45 mins from the coastal jetty in Kuala Dungun. It dimension is about 3 km in size and 2km in width. Nonetheless, this has not prevented it from becoming a paradise for many people particularly diving enthusiasts. The sandy beach blends harmoniously when it comes to the surrounding setting. Additionally, the inland are forested rocky hillsides that make it a scenic location for journey hunters. The hills rising over 800ft offer a panoramic perspective of the island. This rocky surface prolongs from the interior of the island to the ocean and has produced a significant ideal retreat for marine life.
Check out more on Pulau Tenggol Diving.
Pulau Tenggol Diving Package included:
* Accommodation (air conditioned and hot shower room)
* 3 meals a day – last day only breakfast served
* Boat dives with dive guide, filled tank, weights, and weight belt
* Return wooden boat or speedboat transfer
* Boat Dive: 3D2N-4 OR 5 Boat Dive 4D3N-7Boat Dive 5D4N-9Boat Dive
* Surcharge Per Person Per Night (Malaysia and Singapore calendar):
RM 30.00 (Eve & School Holiday) / RM 60.00 (Eve & Public Holiday)
Tenggol Island Beach Resort
The traditional wooden huts of Tenggol Island Beach Resort blends harmoniously when it comes to the atmosphere where it nestles comfortably among the lush eco-friendlies fringing the beach. This comfortable resort is one of the fully-equipped dive outfits on the island, with a full-facility divers' base, oxygen kits for emergencies, top quality tools, and a remarkably well-created divers' kit-up location. There is a Sunlight Fragrance Lunchroom where you get treated to a fragrant genuine Malaysian cuisine. This resort provides the very best services at a sensible rate to make sure that your journey to Tenggol is memorable.
Tenggol Island Resort Package
There are two primary Tenggol Island Resort Package offered by Tenggol Aqua Resort. Both bundles have a minimum keep of 3D2N with alternatives for extension night. On the twin-sharing basis, 3D2N Leisure Bundle prices RM360 each for Coastline Chalets or RM510 per person for Cottage Chalets. The hotel additionally offers 3D2N Diving Package deal that costs RM640 each for Beach Chalets, or RM790 per person for Cottage Chalets. The 3D2N Diving Package deal features five boat dive trips inclusive of air containers and weights only. The full collection of diving tools can be rented out at RM70 per day, which I located somewhat inflated. All package deals are inclusive of return boat transfer from Kuala Dungun and back, three meals per day and selections of the cottage.
Tenggol Island Beach Resort Package
* Snorkelling Package included :
* Accommodation (air conditioned and hot shower room)
* 3 meals a day – last day only breakfast served
* Return wooden boat or speedboat transfer
* Snorkelling Trip : 2D1N – 1 Boat Trip 3D2N-2 Boat Trip 4D3N-3Boat Trip
(With normal mask, snorkel, & life jacket)
From afar, the island resembles an individual setting down, so stood the name 'Tenggol'. 'Tenggak' in Bahasa Malaysia means 'to perch' still, is noticeable as Tenggol in Terengganu's regional language. For several, many years this deserted island maintained its secret location. Aside from local anglers waiting out the weather condition.
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6 Great Group Activities in Richmond, VABy PartyRVA Staff
Sharing experiences with friends is fun. Our team at PartyRVA has compiled our favorite group activities. Make Monday nights fun, make Tuesday nights fun, make any night fun, it's up to you.
The VCUarts Depot building offers a fantastic venue to host everything from corporate meetings and presentations in the Interdisciplinary Studio (pictured) to elaborate, multi-space special events that utilize the whole building.
Originally built in 1907 as a station for the Richmond/Ashland Electric Line, this building served as a transit hub for passengers moving between the state capital and Ashland, Virginia. It was restored in 2013 and in 2014 won the Vision Award from Urban Land Institute.
Wood tones, skylights, hand crafted furniture, historical and architectural features, and up to date technology makes this beautiful space perfect to impress those new clients or to celebrate a milestone.
In addition to the main space, events can take advantage of the space of 2 galleries downstairs, a 10-person conference room, and a state of the art soundstage with motion capture lab and infinity wall, as well as the VCUarts Depot's 3D printers.
Past events have included banquet-style partis including Bar Mitzvahs, Wedding Anniversaries and Receptions, as well as corporate events such as presentations, meetings, trainings, retreats and career fairs.
The maximum capacity for both sit-down and cocktail-style events is 150.
There is a parking lot nearby at the West Broad Street Parking Deck. Street parking is available after 6pm on weekdays and unlimited on weekends. Event parking is available by request, and information can be found here.
The following free add-ons can be made available with a reservation: a projector and screen with full rack, sound and mac mini; two large smart boards with mac minis; seven movable tall tables; 30 stools; 150 chairs. Additional tables are available for rental.
A day-of-event coordinator and/or technologist are available.
If desired, the VCUarts Depot will promote the event through the VCUarts Depot network.
Any caterer is allowed, but must be first approved by the VCUarts Depot's event coordinator. Caterers or the hosting party must hold a license to serve alcohol.
VCUarts Depot events can be scheduled daily from 8am to 12am depending on the University calendar. At least 1 week of lead time is required to schedule events.
Send a message through PartyRVA to receive 10% off the total space reservation cost at VCUarts Depot.
The Depot is available on Saturdays for a $5,000 flat rate for the space.
Additional charges for add-ons apply for additional furniture rentals, and security if serving alcohol.


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Does anyone actually believe this excuse? Just like Conway's excuse for stating Bowling Green Massacre.
Spicer claims he 'clearly meant Orlando' after citing mystery Atlanta terrorist attack three times - The Washington Post
Current Status: Blessed (1)
Seeded on Thu Feb 9, 2017 10:18 AM
keyboard shortcuts: V vote up article J next comment K previous comment


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We Are Pop Slags We're fickle, we're immoral and most important, we like a bit of fun. That makes us pop slags. One day we can't stand you, the next we're all over you. We love pop, so deliver the goods ;-)
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There was an 'emu' wandering around the streets of Doncaster on Thursday.
If you didn't expect to read that today then spare a thought for residents in the South Yorkshire town who woke up to see the large flightless bird wandering down the street.
Police in Doncaster posted photos of the bird on Facebook and, while there is debate as to whether it is an emu or a rhea, who cares - there was a large flightless bird from either Australia or South America kicking about in Doncaster.
Couldn't believe my eyes this morning,... - Doncaster Central LPT | Facebook
Perhaps even stranger is the fact this is the second time this has happened in Doncaster in the past 18 months.
A rhea - this one was confirmed as the South American variety of bird - was on the loose and causing "issues for motorists" in the area of High Melton back in November 2014.
South Yorkshire Police #UK Police are... - Police Alerts UK & News reports | Facebook
The details around today's sighting are not completely clear, but according to a report in the Doncaster Free Press, several of their readers had suggested that today's bird was, in fact, a regular presence in the area.
Apparently a man in the Intake area of the town takes it for walks in the morning...
Of course he does. Emus need to stretch those long legs.


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Camp Fire's Middle School program prepares more than 270 youth annually to achieve success in high school and beyond. Developed in 2011 through a grant from the U.S. Department of Education, the program is currently active in five schools, four in Portland Public Schools and one in Reynolds School District.
Partnerships with the Portland Children's Levy provides funding for three of our five middle school sites. An additional partnership with I Have a Dream Foundation, Oregon deepens our work in at Reynolds Middle School.
Growth Mindset and Sparks
Students are provided with academic support and encouraged to find their "sparks," or passions, that will guide their growth – ranging from creative writing to working with technology. Camp Fire gives students the skills to set personal and academic goals for their future, including finding "spark champions" to achieve their goals.
Our "growth mindset" curriculum encourages students to try new things and remain resilient in the face of obstacles. Activities provided include in-class support, monthly field trips to local businesses and colleges, and regular service-learning projects. Students who participate in Camp Fire demonstrate growth in confidence, leadership, caring for their community, and academic achievement.
Middle School Camp Fire Program Locations:
- Cesar Chavez Middle School, firstname.lastname@example.com
- George Middle School, email@example.com
- Harrison Park Middle School, firstname.lastname@example.com
- Lane Middle School, firstname.lastname@example.com
- Reynolds Middle School, firstname.lastname@example.com


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Home » Home & Hobby » Health & Nutrition » Provillus Review Part-2
Provillus Review Part-2 2.0
- General Info
- Screenshots
- Rate & Reviews
Average Rating:
0.49 Mb
Total: 243 | Last week: 0
Last Update:
11 Mar 2010
Release Date:
10 Mar 2010
This program was not tested against viruses so far. Please check now using this linkDR WEB VIRUS CHECK
Provillus Review Part-2 2.0 Description & Reviews
Only those products for hair loss will be effective which can tackle the true cause of hair loss
Only those products for hair loss will be effective which can tackle the true cause of hair loss. Millions of people around the world get affected by hair loss. It can be cured by many products which are available today. And natural products are always better as there are no side effects. The best products for hair loss are those which are all-natural, dermatologist-formulated solutions which are meant to help slow thinning of hair in both men as well as women. Besides, they should be using natural ingredients that can help stop DHT - the actual cause of hair loss. These products should be able to safely as well as effectively help with thinning hair besides actual hair loss in both men and women. They should be able to stop hair loss besides stimulating healthy hair growth so that you can grow new hair. In case you are using the right kinds of products, you will be able to promote thicker, fuller as well as healthier hair from the follicle. These products can be used to provide cure for male as well as female baldness. There are no side effects when a natural product is used, as all are well aware.
Program Release Status:
Minor Update
Program Install Support:
No Install Support
System Requirements:
Windows 95, 98, ME, 2000, XP, Vista, Windows 7
OS Support:
WinXP, Win2000, Win Vista, Win98, Win7
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FreeSoft4Down.com periodically updates all information about Provillus Review Part-2 2.0 from its publisher, so some information may be slightly out-of-date. You should confirm this information before relying on it. Freesoft4down is a free software download website which provides information and download links as provided by the publisher.Freesoft may use a download manager with offers during the download of the actual software which could be additionally installed with the software being downloaded.Using Provillus Review Part-2 2.0's crack, password, serial numbers, registration codes, key generators, ect. is illegal and prevent future development of Provillus Review Part-2. Download links are directly from our mirror sites or publisher sites, FreeSoft4Down.com does not provide download link for Provillus Review Part-2 2.0 from torrent sites or file hosting sites such as Rapidshare.com, Megaupload.com, Yousendit.com and so on.


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Career Corner: US Military
Adam Karalfa, Staff
June 1, 2015
Filed under Uncategorized
Interested in service the country while earning a competitive wage and receiving great benefits? If so, consider joining a branch of the America's military. To be an enlisted soldier, all that is needed is a high school diploma. With a bachelor's degree or higher, it's possible to become a com...
Honors US History II Decade Projects
Sam Kuhne, Writer
June 1, 2015
Filed under Student Life
Tuesday, May 26th began the Honors U.S. History II decade projects, an assignment that requires students to dress in attire from the 60s, 70s, 80s, or 90s. The week began with a presentation from the 1960's. Members of the class dressed in colorful shirts and as characters from popular 1960's m...


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Alpha Trainer
Alpha Electro
Virus 912
Virus SW 80/100
Sinus 912Sinus FLEX
Taurus M
Taurus electro
Virus SW 121TC EASA
All Pipistrel products are designed and manufactured in our 100% eco-friendly facility and follow our ECOLUTION concept. Learn more about ECOLUTION!
Pipistrel aircrafts
20 Sep 2017Happy Birthday, Finesse Max! Pipistrel has been cooperating with French partners for many years and Finesse Max, our French distributor, is one of our best and most loyal business partners. They were awarded the "Gold Distributor 2015" which is a testimony of their excellence. So it is our pleasure to congratulate Mr. Christian Stuck and his entire team for the 20th anniversary!The Director of Pipistrel's Research & Development institute, Tine Tom...
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01 Sep 2017 Podjetje Pipistrel v sodelovanju s študenti iz treh fakultet razvilo prvo polnilnico za električna letala Read more...
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Quick Facts
- 24 years of experience
- Speaks English
- American University of the Caribbean
- 4 comment
- 2 specialties
- 8 expertises
- 3 locations
Birth Control
Get the facts about birth control so you can decide which type is right for you.
Care Guides
- Birth Control Facts about birth control to help decide which type is right for you.
- Type 2 Diabetes Basic facts about type 2 diabetes & risk factors to be aware of.
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- Flu Facts about influenza (flu), including symptoms and vaccines.
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- GERD Get the facts about gastroesophageal reflux disease (GERD).
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- Home Allergies Facts about indoor allergies, including symptoms & common allergens.
- Chronic Idiopathic Constipation Learn about chronic idiopathic constipation, including treatment
- View All Care Guides Prepare for your next visit with our extensive library of Care Guides
About Dr. Kirk Moss
Dr. Kirk Moss, MD is a Doctor primarily located in Columbus, MS, with other offices in Austin, TX and Addison, TX. He has 24 years of experience. His specialties include Family Medicine and General Practice. He speaks English.
Dr. Kirk Moss has the following 2 specialties
- Family Medicine
A family practitioner is a doctor who specializes in caring for people of all ages, at all stages of life. Rather than focusing on the treatment of one disease or patient population, family practitioners are often the doctors that people see for...
- General Practice
A general practitioner is similar to a family practitioner in that they treat patients of all ages with a variety of health concerns. These doctors treat everyday ailments, some acute and chronic diseases, and also provid health checkups, physicals,...


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EK Success Engraving Tool
Product Actions
To purchase and pick-up in-store, add your selections to your shopping bag. At checkout you can choose whether to pick-up in-store or ship to your home.
BUY ONLINE PICK-UP IN-STORE now available at every location.
Pick-up in one of our locations near you.
Additional Information
Customize your favorite projects with beautiful engravings using the EK Success Engraving Tool. EK Success engraving tools feature a diamond tip that is suitable for engraving on glass, wood, metal, ceramic, stone, plastic, leather, acrylic and more. The stay-on button means that this cordless handheld engraving tool can be used continuously until you turn it off. It runs on four AAA batteries, sold separately.
- Brand: EK Success
- Includes one engraving tool
- Tool operates on four AAA batteries
- Handling Code:
- Web Source: This item is shipped directly from the supplier, please allow an additional 1-3 days processing time.
Standard Delivery
You can usually expect delivery within 6-10 business days from your order date (business days are Monday-Friday, excluding Holidays). You'll receive an email with tracking information when your order is shipped. Please allow up to 5 business days to receive your tracking information.
Some items sent directly from our suppliers may take longer, and will ship separate from the remainder of your order.
Please note that Singer sewing machines cannot be shipped to Guam, Puerto Rico or the U.S. Virgin Islands or to military bases overseas.
Expedited Delivery
Most items will give you the option to upgrade your shipping method for faster delivery. If you choose to upgrade your shipping method, your order must be received by 11:00 AM ET or your order may not be processed until the following business day.
For more details & pricing. See our Full Shipping Policy >
Creativity shouldn't be complicated, so you can return items for FREE any time to our stores for a hassle-free refund with a smile. You can also return online items by mail. See Full Return Policy for Details>


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la la ladybug is doing our inventory and is getting ready for our annual sale. Want to know when the sale starts? Make sure you sign up for the newsletter. Just scroll down to the bottom of the page and click on the "newsletter" link. Subscribers get added discounts too...
Welcome to la la ladybug.
la la ladybug is more than an online store. It's not just about handmade, absolutely adorable, and environmentally friendly products. It's about you - taking care of yourself, your child, and your world. Choosing the finest, hand crafted items for new and expecting parents, we support Canadian small business by providing trendy, responsible products that give you something to brag about.
It truly is trendsetting with a conscience.
Thank you for stopping by, we hope you enjoy your visit.


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Solwold early Northern Sprint Tour points leader. The "hy-per-lube Northern Sprint Tour" has kicked off it's seventh year of bringing the best in wheel banging, dirt throwing, sideways wing to wing Sprint Car racing action to the fans of the...
Solwold early Northern Sprint Tour points leader.
The "hy-per-lube Northern Sprint Tour" has kicked off it's seventh year of bringing the best in wheel banging, dirt throwing, sideways wing to wing Sprint Car racing action to the fans of the great Pacific Northwest and Burlington, WA driver Jason Solwold leads the title race after two events with a total of 160 points.
Solwold, the 1998 series champion finished the 2001 season in third place and is heading into the current season with a feature event win and a third place finish to lead the pack in the points race. Again teamed with the Huson Bros. Racing Team, he is sure to be a top choice for another championship.
Sedro Woolley, WA's Chad Hillier sits in second place with 148 points. Hillier, who chased and won a track championship last year, finished the 2000 NST season in 6th place.
Barry Martinez, of Lynnwood, WA, is in his third year of racing with the hy-per-lube Northern Sprint Tour. Currently in third with 140 points, Martinez finished 9th in 2000 and 6th in 2001.
Defending champion Roger Crockett sits in fourth place with 134 points. Crockett, who moved from Rio Linda, CA to Eugene, OR to be based closer to his car owner, finished a strong second just .024 seconds behind last Saturday's winner and will be a threat to be a repeat champion.
Ladysmith, BC veteran driver Todd Heikes holds down fifth place as he trails Crockett by only 2 points at 132. Heikes, originally from Lake Stevens, WA, saw his best Northern Sprint Tour season in 1998 when he finished in 5th place.
Following the top five spots at this time are 1997 NST champ Rick Fauver of Everett, WA with 120 points in sixth, fellow Everett driver Jayme Barnes has 111 points for seventh and Cottage Grove, OR hot shoe and multi champion Dale Smith is in eight holding 95 points.
Filling out the top ten, it is Travis Hawthorne of Everett, WA with 81 points in 9th and Central Point, OR veteran Bill Nutter trailing in tenth by one point with 80 points.
Action in the 2002 hy-per-lube Northern Sprint Tour will resume on Friday, May 24th at Willamette Speedway in Lebanon, OR and then travel to Southern Oregon Speedway in Medford, OR on Saturday the 25th.
<pre> Pos Car# Name Home Town Points 1 3 Jason Solwold Burlington, WA 2 4C Chad Hillier Sedro Woolley, WA 148 3 66 Barry Martinez Lynnwood, WA 140 4 11 Roger Crockett Eugene, OR 134 5 22T Todd Heikes Ladysmith, BC 132 6 4 Rick Fauver Everett, WA 120 7 9 Jayme Barnes Everett, WA 111 8 75S Dale Smith Cottage Grove, WA 95 9 36 Travis Hawthorne Everett, WA 81 10 3N Bill Nutter Central Point, OR 80 11 V8 Jeff Hodgson St. Albert, AB 77 12 75 Dan Menne Ft. Jones, CA 76 13 69 John Tharp Snohomish, WA 74 14 98 Joe Ramaker Choteau, MT 70 15 12 Tony Menard Nanaimo, BC 66 16 39 Steven Tiner Elverta, CA 64 17 72 Glenn Borden, Jr. Raymond, WA 63 18 2 Scott Aumen Duncan, BC 62 19 21 Ron Myska Green Valley, CA 54 20 3C Cal Herdson Harrison Hot Springs, BC 52 21 5SP Shawna Wilskey Lake Stevens, WA 51 22 0J Jay Cole Shelton, WA 50 23 7 Cale Carder Gold Hill, OR 48 24 57C Chris Schmelzle Victoria, BC 43 25 41H Danny Horner Cottage Grove, OR 41 26 18 Travis Jacobson Lake Stevens, WA 37 27 27C Jeff Jansma Clear Lake, WA 28 28 23 Richie Peterson Great Falls, MT 21 29 34 Don Fry Montesano, WA 18 30 22 Geoff Beck Morinville, AB 15 31 77F Jason Finley Ft. Jones, CA 12 32 14 Robert McNulty Mt. Vernon, WA 11 33 28 Rick Smith Mt. Vernon, WA 9 34 34 Meilssa Yates Roseburg, OR 6 35 17I Richie Brown Alvadore, OR 4 36 49 Dawn-L Morrison Millet, AB 4 37 99 Tyler Pincemin Morinville, AB 2 38 9H Mark Hinkle Scappoose, OR 2


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Textbook on International Human Rights / Edition 5
Paperback - Rent for
Textbook on International Human Rights / Edition 5
"This textbook provides a manageable text in Human Rights, covering all major aspects. The origins of contemporary human rights are discussed and a detailed analysis of the international system under the auspices of the United Nations is provided. It addresses the scope and enforcement of international human rights and highlights the advantages and disadvantages of the present system - the Universal Declaration and the International Covenants are considered, providing a link to the following detailed analysis of the Council of Europe. The institutions and processes are considered and a detailed tabulation of the rights, as interpretated by the EU courts, is provided."
Product Details
|Oxford University Press
|Publication date:
|Edition description:
|Older Edition
|Product dimensions:
|6.70(w) x 9.60(h) x 1.00(d)
Table of Contents
1. Introduction
Customer Reviews
Average Review


text:
I cannot walk away from your purring and crunching
Though it turns my stomach to hear the joy in your brunching
On blood and marrow slurped and sucked as little bones crack
And a new mouse-tailed whisker from you sprouts.
These predatory laws of behavior elude me.
Why torment and tease and play with life?
And, my God, did that mesmerized mouse just run back to you?
When you offered it life? Is prey in love with death? What lies did you tell it?
I tuck in my chin and defend against an uppercut to my spirit.
My elbows guard her solar plexus from having breath kicked out of her.
Upon the balls of my feet she rises light and nimble
To take the center of the ring and not a corner.
Would I could sit cross legged on a water lily,
But for me awareness comes on the blade of a knife.
And now I see the revolutions of it falling and grasp it by the handle.
My awareness is not the awakening we all are seeking, perhaps.
I ponder this as my cat brings me yet another morbid gift to contemplate.
Sunday, November 18, 2012
Morbid Gifts to Contemplate
2 comments:
I so relate to this! Difficult when beloved critters show their animal natures. We'd rather they sit daintily on puffy pillows - or at least not have to look at that tail slithering down the throat, or have to remove the corpses from the bathmat! Love it.ReplyDelete
I've tried a puffy pillow. It was shredded by the dog! He was probably wishing it was the cat, but knew he wouldn't get away with it so he thrashed the pillow instead.ReplyDelete
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text: Are you looking for an affordable revenue or starter home in central Ponoka? Here is your chance! This cozy, 1.5 story home is centrally located and within easy walking distance to the downtown area, walking trails, parks, and schools. The standard size lot is also zoned R4 and would make an excellent candidate for a multi-family dwelling.


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Monday, May 22, 2017
Dems are gunning for Trump — here's how he can survive
NYPost by Micheal Goodwin
Reader Freda Barry objects to my view that President Trump's errant words and deeds are testing the faith of his followers. She writes: "No, supporters of President Trump are not losing faith. We are, however, totally dismayed by the unrelenting attacks on our president from the corrupt media!
"Please don't give up on us or the president we VOTED into office. We still pray that he will NOT give up, even under continuous assault."
Her plea is obviously heartfelt, and others also invoked prayer in delivering similar messages. But while I haven't given up on Trump, I am concerned about the state of his presidency.
Indeed, events in recent days added to my worry.
So much so that I believe Trump, having twice called the special-counsel investigation a "witch hunt," must now discipline himself to avoid getting burned at the stake.He pulled off one of the great political upsets in history to get to the White House, but may need another miracle to finish what he started.
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French Fries
- Ian Viggars
Golden, crispy on the outside just the way we like 'em. hi-rise french fries! A3 / A4 C-Type print on Fuji Crystal Archive paper
Ian Viggars is an East London Artist who paints & illustrates food in a Pop Art style. This print features in our curated show Pop (t)Art by Ian Viggars at People's Park Tavern Sep- Oct 2016
If ordering a framed print please allow 5-7 days + shipping time /or email us before purchase to arrange collection from our E1 store (or if buying more than one print) so we can ammend /or combine the shipping cost


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In a lonely September garden, I sit with my cup of coffee, My cognac and my book, Turning over deep thoughts. The wind blows through The pages of my book With the smell of grass. Riffling the pages quickly, Muddling the letters In front of my eyes, Who knows how many Thousands of words Both small and great. Are lost in the wind. I hastily close the cover, Pressing it firm with my hand; Peeking, see all is in order In a lonely September garden, With a cup of coffee, A book and a cognac.
Metin ALTIOK Translation : Angela ROOME


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Raleigh newspaper carrier 'in shock' after being shot at on route
Posted January 12
Raleigh, N.C. — A veteran newspaper carrier was approached by four men and shot at while he was completing his route on Durant Road on Tuesday morning.
The man, who did not want to be identified, said he was approached by a car carrying four men near the train tracks when someone got out of the car and started shooting at him.
In a 911 call, the man can be heard telling the dispatcher that the shots were fired from a small blue car being driven by a man in a white hoodie. He said he wasn't sure why the shots had been fired, but speculated that perhaps the driver of the blue car was upset over the fact that their car was on the icy part of the road while the car belonging to the newspaper carrier was on a cleared portion of road.
The man's wife said there were multiple bullet holes in his car.
"He put the car in reverse and bullets just started coming at the windshield," she said. "It hit his tire, it hit the front bumper. So, he got out of there pretty fast."
Both the newspaper carrier and his wife say they are in shock.
"We were shocked. It's shocking. He's still in shock right now," she said.
Another incident with a similar suspect description was reported on Thornton Commons Drive and Meryton Park Way the same morning, police said. In that case, police said a suspect pointed a gun at the carrier's car but didn't shoot. In a 911 call, the driver said that four people jumped out of a small, four-door black car and pointed the gun.
Investigators believe the carriers were specifically targeted.
"He's been doing this for 20 plus years. This never happens," his wife said.
Police and the victims are urging other newspaper carriers to take precautions.
"Be cafe, be safe, be aware of your surroundings," she said.
Both carriers work for the News and Observer. A spokesperson for the newspaper said they are "reaching out to all of the carriers urging them to be alert and cautious."
The spokesperson said the carriers to not handle cash.
No arrests have been made.


text: Want to own a fast, powerful, and flexible media player for your PCs or Tablets? Zoom Player Max from Inmatrix.com is the wise choice for you. It plays almost any format you throw at it and its interface is extremely intuitive and easy to use. Main Features: No bloat, intuitive interface with a user-friendly design Skins for every resolution, including 4K screens Install Center application, capable of detecting and securely installing up-to-date decoders, missing system components and language packs to improve stability and versatility DVD Playback from Disc, Hard Drive or network connection Quickly jump between multiple saved positions using media Chapters Media Library JukeBox mode meta-data scraping for Movie/TV/Music posters, backdrops, still images, title, description, genre, cast and more! Media Library YouTube (Channels/Searches/Trends) integration Media Library RSS feed reader ….and many more Price: $29.99 HOW TO GET ZOOM PLAYER MAX GIVEAWAY? Zoom Player is the most sophisticated, flexible and powerful Media Player. Without spending $29.99 for Zoom Player MAX, you will get a free license key for Zoom Player MAX if take the following steps: 1. Go to this GIVEAWAY PAGE 2. Enter your email address 3. After clicking to "SIGNUP", a new tab contained download link will be shown. 4. Download the setup file "zp1350max_giveaway.exe" and install it Note: You should uncheck "Run the Install center" as below during installation And this is your giveaway version of Zoom Player MAX v13.5 too many skins https://www.inmatrix.com/zplayer/skins/ This version is a non-upgradable version for free Special Offers for you: To buy paid-version of Zoom Player MAX at a low cost, don't let any Inmaxtrix coupon codes pass you by. Take some minutes to check the coupon codes to save up to 40% on Zoom Player MAX.


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Business TypeTrading Company
Main MarketsNorth America South America Western Europe Eastern Europe Ea
Product/ServiceCocoa beans, kernel nut, teak,
Business OwnerTia Samson
Employees5 - 10 People
RegionCote D'Ivoire
CategoryBusiness Services - Agency Services
Company Tags
Company Introduction
We are specialized in tropical products. We have more than 30,000 farmers which belongs to our association. Since 1998, our company has signed partnership contract with many company here in our country.
Contact Information
Contact PersonMr. Tia Samson
Fax Number225-23-458713
Post Code
AddressBP 202 Post Entreprise Abidjan Cedex 1, Abidjan, Cote D'Ivoire
Online MapProduitropicaux Map & Directions


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The B.S. degree in Informatics is administered by a faculty committee with a rotating membership.
- Personalized learning, social media, learner analytics, mobile learning, cognitive science, usability testing, human-computer interaction
- Data mining, web and text mining, data cleaning and information extraction, workflows and semantic web services, database systems
- Adaptive technology, personalized learning, interactive educational systems
- Game design, game development, digital fabrication, AI in design, virtual reality, building information modeling, procedural modeling, computer graphics, PRISM Lab at ASU
- Learning theory, instructional design, educational technology, game-based learning environments, simulations, multimedia, collaborative learning
- Undergraduate program lead, Informatics Applications of AI to education (Intelligent Tutoring Systems; teachable agents; tutorial NL dialogue systems); human learning (student modeling; cognitive modeling); cognitive science
- Intelligent tutoring systems, computer-supported collaborative learning, online learning communities, tangible embodied environments, human-computer interaction