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The recognition of a C-terminal motif in E. coli SecM (150FXXXXWIXXXXGIRAGP166) inside the ribosome tunnel causes translation arrest, but the mechanism of recognition is unknown. While single mutations in this motif impair recognition, we demonstrate that new arrest-inducing peptides can be created through remodeling of the SecM C-terminus. We found that R163 is indispensable, but that flanking residues that vary in number and position play an important secondary role in translation arrest. The observation that individual SecM variants showed a distinct pattern of crosslinking to ribosomal proteins suggests that each peptide adopts a unique conformation inside the tunnel. Based on the results, we propose that translation arrest occurs when the peptide conformation specified by flanking residues moves R163 into a precise intra-tunnel location. Our data indicate that translation arrest results from extensive communication between SecM and the tunnel and help explain the striking diversity of arrest-inducing peptides found throughout nature.
During translation nascent polypeptides traverse a long aqueous tunnel in the large ribosomal subunit before emerging into the cytoplasm (Milligan and Unwin, 1986; Yonath et al., 1987). The ribosome tunnel has a length of ~100 Å from the peptidyl transferase center (PTC) to the exit site and an average diameter of ~15 Å (Nissen et al., 2000). The tunnel is highly irregular in shape and contains numerous grooves and cavities. While the surface of the tunnel is comprised mainly of 23S rRNA, it also contains non-globular segments of several ribosomal proteins. Two of the proteins, called L4 and L22 in bacteria, reside near the tunnel entrance, and a third protein, L23, is located near the exit. A narrowing of the tunnel ("constriction point") is seen ~30 Å from the PTC where a conserved β hairpin loop of L22 is in close proximity to L4.
Although the functional significance of the ribosome tunnel is poorly understood, there is growing evidence that it is not simply a passive conduit. Early studies showed that ribosomes protect ~30–40 amino acids of a typical nascent chain (Malkin and Rich, 1967) and thereby suggested that polypeptides transit the tunnel in an extended conformation (~3.5 Å /residue). Recent work, however, has indicated that at least some polypeptides adopt a more compact conformation inside the tunnel (Hardesty and Kramer, 2001; Lu and Deutsch, 2005; Woolhead et al., 2004 and 2006). Moreover, specific polypeptides that are too short to emerge from the ribosome tunnel have been shown to arrest translation in both prokaryotes and eukaryotes (Tenson and Ehrenberg, 2002). This cis-acting translational attenuation serves to regulate the expression of a downstream gene and often requires a threshold concentration of a small molecule such as an amino acid or antibiotic. For example, a 24 amino acid leader peptide encoded by the E. coli tnaC gene inhibits its own termination in the presence of tryptophan and thereby activates the transcription of a downstream operon (Gong and Yanofsky, 2002). Likewise, peptides encoded in small open reading frames located at the 5' end of fungal carbamoyl phosphate synthetase (the arginine attenuator peptide, or AAP), plant cystathionine γ-synthase, and mammalian S-adenosylmethionine decarboxylase mRNAs stall their own translation in response to high concentrations of arginine, methionine and polyamines, respectively (Tenson and Ehrenberg, 2002). Curiously, the ribosome-stalling polypeptides identified to date vary considerably in length and share no sequence homology.
Physiological signals that do not involve small molecules can also trigger translation arrest. In E. coli, a secreted 170 amino acid protein called SecM regulates the translation of the co-transcribed secA gene in response to the secretion status of the cell (Oliver et al., 1998). Under normal conditions, the SecA Shine-Dalgarno sequence is masked by secondary structure in the secM-secA mRNA. Transient stalling of the ribosome at P166 destabilizes the structure and facilitates the synthesis of a basal level of SecA. The stalling is released by a "pulling" force exerted by the Sec machinery on SecM (Nakatogawa and Ito, 2001; Butkus et al., 2003). In contrast, under secretion-impaired conditions, a prolonged translation arrest increases the exposure of the SecA Shine-Dalgarno sequence and leads to overproduction of the protein (Nakatogawa and Ito, 2001). Consistent with the notion that stalling occurs in the ribosome tunnel, a short segment derived from the C-terminus of SecM can function autonomously to inhibit the translation of unrelated downstream sequences (Nakatogawa and Ito, 2002).
There is considerable evidence that translation arrest is initiated by the interaction of specific nascent polypeptide sequences with tunnel components. Mutations in L4, the L22 β hairpin loop and segments of 23S rRNA that line the top third of the ribosome tunnel impair SecM- and TnaC-mediated translation arrest (Nakatogawa and Ito, 2002; Cruz-Vera et al., 2005; Lawrence et al., 2008). Furthermore, despite their vast sequence diversity, all arrest-inducing peptides examined to date have been shown to contain one or more specific residues that play a critical role in regulating translation (Alderete et al., 1999; Fang et al., 2000; Vazquez-Laslop et al., 2008). Mutation of any of the nine key residues in the E. coli SecM motif 150FXXXXWIXXXXGIRAGP166 (which are located at the top of the tunnel at the time of ribosome stalling) reduces translation arrest activity (Nakatogawa and Ito, 2002). Likewise, W12, D16, and the terminal P24 residue are essential for TnaC-mediated arrest (Cruz-Vera, et al., 2005 and 2008). The mutational data suggest that the recognition of specific motifs inside the ribosome tunnel transmits a signal outward that ultimately inhibits further translation. Consistent with this hypothesis, an analysis of SecM-stalled ribosomes by cryo-EM revealed a dramatic change in ribosome structure (Mitra et al., 2006). Presumably due to conformational changes near the PTC that inhibit the transpeptidation reaction, P166-tRNA remains bound to the A site and is not incorporated efficiently into the SecM nascent chain during translation arrest (Muto et al., 2006).
Beyond this basic phenomenology, the mechanism by which specific nascent polypeptide sequences are recognized in the ribosome tunnel remains obscure. The observation that the spacing between critical residues in arrest-inducing peptides cannot be altered (Gong and Yanofsky, 2002; Nakatogawa and Ito, 2002) suggests that translation arrest might entail the interaction of the nascent chain with multiple sites along the tunnel. In this regard it is intriguing that TnaC, SecM and AAP all have an essential residue 11–12 amino acids from the arrest point. Because many mutations that impair translation arrest affect components of the ribosome around the constriction point (where this key residue would be located if the nascent chain were in an extended conformation), it has been proposed that the narrow part of the tunnel might be a "discriminating gate" (Nakatogawa and Ito, 2002). Consistent with this hypothesis, a recent study showed that antibiotic-dependent ribosome stalling is sensitive to mutations in the same region (Vazquez-Laslop et al., 2008). It is unclear, however, how unrelated sequences can all be deciphered at the constriction point. Even in the simplest scenario, the observation that mutations in L4, L22 and neighboring rRNA segments differentially affect SecM-, TnaC-, and antibiotic-dependent ribosome stalling suggests that sequence discrimination occurs in an elaborate binding site. To complicate matters, a recent study showed that ribosomes induce the SecM C-terminus to adopt a compact conformation during translation arrest and thereby suggest that the recognition of specific sequence motifs involves a complex series of events (Woolhead et al., 2006).
To gain insight into the mechanism of peptide recognition inside the ribosome tunnel, we explored the degree to which the SecM translation arrest motif tolerates sequence variation. We hypothesized that if mutations that impair arrest perturb critical interactions between the ribosome and the nascent chain, then it might be possible to isolate second-site suppressor mutations that either restore these interactions or strengthen other interactions. In this way we hoped to clarify the nature and number of interaction sites. Surprisingly, we found that extensively remodeled motifs containing as few as three of the nine key residues in the E. coli SecM motif are completely functional. Further analysis suggested that a single invariant residue (R163) is "recognized" by the ribosome, while other more variable residues primarily influence the conformation of the nascent chain in the tunnel. Based on the data, we propose that translation arrest occurs when the conformation of nascent SecM facilitates the positioning of R163 at a specific intra-tunnel site and that mutations near the constriction point only indirectly affect peptide recognition. In addition, we propose that, like SecM, all arrest peptides consist of both essential and context-dependent sequence elements, and that this feature helps to explain their diversity.
Although alanine-scanning mutagenesis indicated that only residues 150, 155–156 and 161–166 of E. coli SecM [SecM(Eco)] are required for complete translation arrest (Nakatogawa and Ito, 2002), we found that the mutation of residues 157–160 to proline also causes severe translation arrest defects (Woolhead et al., 2006). This observation suggested that constraining the polypeptide backbone between residues 157–160 might perturb the positioning of one or more key residues inside the ribosome tunnel and thereby prevent recognition of the translation arrest motif. We reasoned that second-site suppressor mutations might be isolated that would either restore the proper positioning of these key residues or introduce conservative substitutions at key positions that would restore recognition. To test this hypothesis, we used an assay based on the finding that the fusion of the SecM(Eco) arrest motif to LacZα inhibits synthesis of the reporter, but that mutations that impair translation arrest restore LacZα production and turn colonies dark blue on indicator plates (Nakatogawa and Ito, 2002). We randomly mutagenized the SecM moiety of a SecM-LacZα fusion carrying the Q160P mutation, which impairs arrest, and screened for pale blue colonies (white colonies contained nonsense or frameshift mutations). Ultimately we obtained six plasmids designated Sup1-Sup6 that produced colonies of the desired color and that encode one or more second-site mutations (Fig. 1A).
We confirmed that the suppressor mutations isolated in the genetic screen restore SecM function using an in vivo translation arrest assay. In these experiments a secretion-deficient E. coli strain (CU164) was transformed with a plasmid harboring full-length SecM(Eco) (pSecM), pSecM containing an arrest-compromising mutation (Q160P or P166A), or pSecM containing one of the suppressor sequences. Cells were subjected to pulse-chase labeling, and SecM was immunoprecipitated. Consistent with previous results (Nakatogawa and Ito, 2002), only the translation-arrested fragment of wild-type SecM (166 residues) was observed; no full-length protein (170 residues) was detected even after a 2 min chase (Fig. 1B, lane 1–3). As also previously observed, the P166A mutation abolished translation arrest and led to the production of only full-length SecM (Fig. 1B, lanes 25–27). The Q160P mutation was less severe in that about half of the pulse-labeled SecM was isolated as the translation-arrested fragment. After a 2 min chase, however, only full-length protein was observed, suggesting that the mutation destabilizes the translation-arrested state (Fig. 1B, lanes 4–6). Interestingly, Sup1–3 yielded a single polypeptide that co-migrated with the stalled fragment of wild-type SecM(Eco) (Fig. 1B, lanes 7–15). The identity of this band as a translation-arrested product was verified using a coupled transcription-translation assay (Fig. S1A). These results indicate that the second-site mutations in Sup1–3 completely restore translation arrest activity. Sup4–6 only partially restored activity in vivo in that only ~50–60% of the SecM was isolated as a translation-arrested fragment after a 2 min chase (Fig. 1B, lanes 16–24 and Fig. S1A).
Surprisingly, only some of the suppressor mutations appeared to support our original hypothesis. Sup5 and Sup6 harbor mutations in the "non-essential" residues 157–159 that might restore the proper positioning of key amino acid side chains. In addition, Sup4 contains a modest mutation of a key residue (I156T) that might promote translation arrest in the context of a nascent polypeptide constrained by the Q160P mutation. Sup4–Sup6, however, are only relatively weak suppressors. All of the strong suppressors (Sup 1–Sup3) contain at least one highly non-conservative alteration of a key residue (G161P and, in the case of Sup3, W155E). The observation that a severe alteration of residue W155 can be compatible with translation arrest is especially striking because W155 has been predicted to be located near the tunnel constriction point (Berisio et al., 2003). The isolation of suppressors lacking elements of the canonical SecM(Eco) arrest motif provided the first indication that at least some of the key residues might not be involved in direct interactions with the ribosome and that the constriction point might not be a "discriminating gate".
Interestingly, the C-terminal sequences of Sup1–Sup3 closely resemble those of uncharacterized SecM homologs found in the genomes of Pasteurellales (van der Sluis and Driessen, 2006). Like the SecM homologs found in Enterobacteriales, these homologs are completely divergent in overall sequence and length, but they are directly upstream of secA and their C-terminal sequences can be aligned (Fig. 2A). Like Sup1–3, Haemophilus ducreyi and Pasteurella multocida SecM homologs both contain prolines at positions that are equivalent to Q160 and G161 of SecM(Eco). These homologs also contain the same sequences as Sup2 and Sup3 at positions that are equivalent to W155 and I156 of SecM(Eco).
The fact that the outcome of our genetic screen seemed to mimic the natural evolution of SecM raised the possibility that C-terminal sequences of divergent SecM homologs might stall translation in E. coli. To test this hypothesis, we replaced the last 22 residues of SecM(Eco) with moderately divergent (5/9 key residues conserved) and highly divergent (3/9 key residues conserved) C-terminal SecM sequences derived from from Haemophilus influenzae or Mannheimia succiniciproducens (Fig. 2B). CU164 transformed with plasmids encoding the chimeras [SecM(C-Hi) and SecM(C-Ms)] or the chimeras containing the P166A mutation were subjected to pulse-chase analysis, and the chimeric proteins were immunoprecipitated. Remarkably, SecM(C-Hi) and SecM(C-Ms) produced a single polypeptide that co-migrated with the translation-arrested fragment of SecM(Eco) (Fig. 2C, lanes 7–9 and 13–15). In contrast, SecM (C-Hi) and SecM (C-Ms) containing the P166A mutation yielded products that co-migrated with full-length SecM (Fig. 2C, lanes 10–12 and 16–18). We confirmed that these polypeptides correspond to translation-arrested and full-length versions of SecM, respectively, using coupled in vitro transcription-translation assays (Fig. S1B). We also found that the native H. influenzae SecM protein undergoes elongation arrest in E. coli cell-free extracts (Fig. S1C). These data show that distantly related SecM homologs are completely functional in E. coli despite only weak sequence conservation.
Based in part on the observation that P166 plays an essential role in SecM-mediated translation arrest even when the arrest motif is substantially altered, we conjectured that Sup1–6, SecM(C-Hi) and SecM(C-Ms) stall translation by the same mechanism as SecM(Eco). To test this idea, we first compared the position at which stalling occurs in E. coli SecM and each of the SecM variants. Coupled transcription-translation reactions were programmed with full-length SecM DNA templates and peptidyl-tRNAs were resolved on low pH Tris-acetate gels (Kirchdoerfer et al., 2007). Northern blot analysis was conducted using oligonucleotide probes complementary to tRNAGly and tRNAPro , the tRNAs that would be linked to the nascent chain if stalling occurs at residues 165 and 166, respectively. Because residue 165 in SecM(C-Ms) is a serine instead of a glycine, we also used a probe that can detect tRNASer. Consistent with previous results showing that G165 is the terminal residue of stalled SecM(Eco) (Garza-Sanchez et al., 2006; Muto et al., 2006), we found that a significant fraction of the SecM(Eco) nascent chain was attached to tRNAGly (Fig. 3, top panel, lane 1). Interestingly, we found that all of the variant SecM nascent chains were also attached to tRNAGly except for SecM(C-Ms), which was attached to tRNASer (Fig. 3, top and middle panels). While we found that a fraction of each stalled product contained tRNAPro (Fig. 3, bottom panel), our results are consistent with the observation that a subset of stalled SecM(Eco) products contain P166 in vivo (Collier et al., 2004; Sunohara et al., 2004). Furthermore, we found that like SecM(Eco) RNCs, SecM(C-Hi), SecM(C-Ms) and Sup 1–6 RNCs were resistant to attack by puromycin, presumably because access to the A site is blocked by the presence of tRNAPro (Woolhead et al., 2006; Muto et al., 2006) (Fig. S2). Taken together, these data provide evidence for a universal SecM stalling mechanism that is independent of variation in the arrest peptide sequence.
The results described above show that the SecM translation arrest motif tolerates considerable sequence variation. While the data indicate that P166 cannot be altered, this amino acid has a unique role in the arrest process as an aminoacyl tRNA, and it never enters the ribosome tunnel. The plasticity of the SecM sequence N-terminal to P166, however, challenges the idea that translation arrest results from the precise positioning of most or all of the key residues in the SecM(Eco) motif at fixed recognition sites located along the ribosome tunnel and instead suggests a more complex interaction between the nascent chain and the tunnel wall. To gain further insight into the relationship between sequence and function, we performed alanine/serine scanning mutagenesis on residues 149–165 of SecM(C-Ms) to identify residues that are required for translation arrest.
Interestingly, we found that the mutation of only one residue, R163, completely abolished translation arrest mediated by SecM(C-Ms). R163 is one of only three amino acids that is invariant in all SecM homologs sequenced to date, and mutation of this residue also abolishes translation arrest mediated by SecM(Eco) (Nakatogawa and Ito, 2002; Fig. S3A). Mutation of R163 to other amino acids (including lysine) abolished translation arrest activity in vivo, but changing the CGC codon to synonymous arginine codons had no effect on function (Fig. 4A, lanes 46–48 and Fig. S3B, lanes 4–24). Shifting R163 -1 or +1 in the SecM(C-Ms) sequence also resulted in the production of only full-length protein (Fig. S3B, lanes 31–36). These results demonstrate that both the identity of arginine at residue 163 and its exact position in the amino acid sequence play a decisive role in SecM-mediated translation arrest.
Beyond the common requirement for R163, we found a dramatic disparity in the role played by other amino acids in the arrest mediated by SecM(Eco) and SecM(C-Ms). Most notably, while the alteration of residues 159–166 in SecM(C-Ms) impaired elongation arrest activity in vivo, mutation of residues 149–158 had no effect (Fig. 4A). This is a striking result because F150, W155 and I156 all play a very significant role in the stalling mediated by SecM(Eco) (Nakatogowa and Ito 2002; Fig. 5, lanes 10–18). The data show that while the SecM(Eco) arrest motif spans 17 amino acids, the SecM(C-Ms) arrest motif spans only 8 residues. Moreover, we found that the relative contribution of each residue between 159 and 166 to translational stalling is different in SecM(Eco) and SecM(C-Ms) (Fig. 4B). For instance, H159 and A160 play a modest role in SecM(C-Ms)-mediated translation arrest, but the equivalent residues in SecM(Eco) do not appear to play any role. In addition, mutation of P161 in SecM(C-Ms) strongly impairs translation arrest activity, but mutation of G161 in SecM(Eco) has only a minor effect. These differences strongly suggest that all SecM arrest motifs contain a class of residues that play a secondary role in translational stalling, and that the number, position and identity of these residues is context-dependent (i.e., dependent on the overall sequence of the C-terminal peptide). Consistent with this interpretation, A164 in SecM(Eco) is required for complete arrest but, conversely, the presence of an alanine at this position in SecM(C-Ms) causes an arrest defect. Because the secondary residues are so variable, it seems unlikely that they contribute directly to the transmission of a signal through the ribosome tunnel.
An examination of the Sup1–3 sequences as well as of the natural evolution of SecM suggested a plausible explanation for the existence of relatively short translation arrest motifs. Although E. coli and its close relatives require a glycine at position 161, Sup1–3 and the SecM homologs of Pasteurellales all contain a proline at this position (Figs. 1A and and2A).2A). In addition, the sequence N-terminal to P161 is not conserved in the Pasteurellales. Finally, while the Q160P mutation severely impairs arrest mediated by SecM(Eco), a proline is found at position 160 in Sup1–3 and in some of the Pasteurellales homologs. The sequence information suggests that prolines at positions 160 and 161 might reduce the role of F150, W155 and I156 in translation arrest. To test this hypothesis, we combined the F150A, W155A and I156A mutations with either the G161P mutation or the double Q160P/G161P mutation and examined the effect of the mutations on SecM(Eco)-mediated translation arrest in vivo. Consistent with our hypothesis, we found that the defect produced by the W155A/G161P and I156A/G161P double mutations was similar to that produced by the G161P mutation alone but was markedly less severe than that produced by either W155A or I156A alone (Fig. 5A, lanes 13–30). Furthermore, when residues 160 and 161 were both converted to proline, F150, W155 and I156 could all be mutated to alanine (either singly or in combination) without producing any discernable arrest defect (Fig. 5A, lanes 34–42). R163, however, was still strictly required for arrest activity (Fig. S3A lanes 19–21). The observation that a +1 shift of Q160P/G161P completely abolished translation arrest indicates that the position of the prolines is important (Fig. 5A, lanes 31–33). Taken together, the data suggest that prolines at positions 160 and 161 reduce the arrest–inducing peptide to the simple core motif 162IRXXP166 (where X is a small amino acid).
Based on the finding that SecM arrest peptides contain residues that appear to provide contextual information and that prolines profoundly influence the size and functionality of arrest motifs, we conjectured that divergent arrest peptides might adopt unique conformations inside the ribosome tunnel (or might be situated in distinct positions relative to the tunnel walls). Presumably such unique configurations would result from inherent constraints (e.g., the presence of prolines), interactions between the nascent polypeptide and tunnel components, or constraints imposed on the polypeptide by the tunnel environment. Consistent with the idea that the conformation of the nascent chain is relevant to arrest induction, proline mutations at positions 158 and 160 of SecM(Eco) each cause strong arrest defects, but the combination of the two prolines (in Sup4) almost completely restores translational stalling (Fig. 5B). The simplest explanation of this result is that the two prolines cancel out the contribution that they each make individually to polypeptide conformation.
We used a site-specific photocrosslinking approach to obtain more direct information about the disposition of SecM arrest peptides inside the ribosome tunnel. We modified an amber suppression system involving the co-expression of an amber suppressor tRNA and an amino acyl-tRNA synthetase from M. jannaschii that are orthogonal to the E. coli translation machinery (Xie and Schultz, 2006). While this method has previously only been used to incorporate "unnatural" amino acids into proteins in a site-specific fashion in vivo, we adapted the technology for use in cell-free transcription-translation reactions. Reactions were programmed with full-length SecM(Eco), Sup1 and SecM(C-Ms) DNA templates containing single amber mutations. Sup1 was chosen for these experiments because it differs from SecM(Eco) at only two positions, and SecM(C-Ms) was chosen because it is the most divergent SecM variant that we analyzed. Amber suppression led to the incorporation of photoreactive p-benzoyl-L-phenylalanine (Bpa) or p-azido-L-phenylalanine (Azp) into the protein. To reduce the frequency of termination at the amber codon and thereby enhance the efficiency of amber suppression, we made extracts from a strain that contains the release factor 1 (RF1) mutant allele prfA1 (Zhang et al., 1996). Based on the ratio of arrested SecM to SecM that was terminated at the amber codon, the incorporation efficiency was ~10–20% (Fig. 6, left panel, compare "arrested" to "no suppression"). Although the incorporation of amino acid analogs at some "non-essential" positions abolished translation arrest (Fig. S4), the incorporation of Bpa or Azp at residues 141, 144, 149, 151, 152, and 158 did not impair the function of the arrest peptide (except as noted below). After the cell-free reactions were completed they were exposed to UV light and immunoprecipitations were conducted with antisera raised against L4, L22 and L23 to assess the proximity of each photoprobe to tunnel components.
Interestingly, we found that each SecM variant exhibited a distinct pattern of crosslinking to ribosomal proteins. When Bpa was incorporated into Sup1 and SecM(C-Ms) at residue 141, photoadducts of ~29–30 kD were clearly visible in the UV-irradiated samples (Fig. 6A, lanes 10–12). These bands corresponded to SecM (~18 kD) crosslinked to roughly equal amounts of L22 and L23 (~11–12 kD) (Fig. 6A, right panel). Smaller amounts of these crosslinked products were observed in reactions programmed with SecM(Eco), but the disparity corresponds to a reduction in the efficiency of translation arrest (Fig. 6A, lane 1). The incorporation of Bpa at residue 144 did not affect translation arrest, and SecM variants containing the photoprobe at this position were all crosslinked to L22 with similar efficiencies (Fig. 6B). The crosslinking of sites in the nascent chain that are ~22–25 residues from the PTC to L22 and L23 is consistent with previous results (Houben et al., 2005) and suggests that the proximity of each SecM variant to the tunnel wall is similar near the distal end of the tunnel. Striking differences between the variants were observed when the probes were placed closer to the PTC, however. Incorporation of Bpa or Azp at residue 149, 151 or 152 resulted in significant crosslinking between SecM(C-Ms) and L22 and/or L4 (Fig. 6C–E). Crosslinking between Sup1 and the same ribosomal proteins was observed at all of these positions, but at positions 151 and 152 the signal was much weaker. In contrast, no crosslinking was observed when the photoprobe was incorporated at residue 149 or 152 of SecM(Eco) (position 151 cannot be evaluated because Bpa perturbs translation arrest). Finally, crosslinks were observed between Sup1 and SecM(C-Ms) containing Azp at residue 158 and L4, but not between the corresponding SecM(Eco) derivative and L4 (Fig. 6F). Taken together, these results strongly suggest that the segment of each of the three SecM isoforms adjacent to the arrest motif core has a distinct conformation (or spatial relationship to the tunnel walls) that might extend into the core itself. Because the Sup1 crosslinking pattern is "intermediate" between those of SecM(Eco) and SecM(C-Ms), it appears that conformational differences between two polypeptides correlate with the degree to which their sequences diverge.
The notion that SecM variants adopt unique conformations that result at least in part from interactions with tunnel components predicts that changes in tunnel structure would produce a variable effect on the activity of each variant. Consistent with our prediction, we found that mutations in both L22 and 23S rRNA impaired the arrest mediated by the SecM variants to different degrees (Fig S5).
In this report we show that SecM mutants isolated in a genetic screen as well as divergent SecM homologs mediate translation arrest in secretion-deficient E. coli as effectively as SecM(Eco). The identification of fully functional arrest peptides that contain as many as six mutations in the arrest motif 150FXXXXWIXXXXGIRAGP166 was counterintuitive because single mutations reduce the efficiency of translation arrest. Despite the striking sequence diversity of the SecM variants, they all appear to mediate translational stalling by the same mechanism. Interestingly, we found that while mutation of R163 and P166 abolished the translation arrest mediated by both SecM(Eco) and the most divergent SecM variant, SecM(C-Ms), partial arrest defects resulted from the mutation of other C-terminal residues that vary in position, number and side chain chemistry. Furthermore, photocrosslinking data provided evidence that the sequence variation that we observed correlates with a distinct positioning of the C-terminus of individual SecM variants relative to L4 and L22. Because the presence of proline residues (which often impose conformational constraints) strongly affects arrest activity, it seems likely that the photocrosslinking data reflect conformational differences among the nascent polypeptides. Additional studies will be required, however, to determine the degree to which the structure of each nascent chain (as opposed to the structure of the tunnel) varies. In any case, the observation that individual nascent polypeptide chains are situated differently inside the ribosome tunnel strongly suggests that it provides an interactive, rather than a passive, environment.
The remarkable plasticity of the SecM arrest motif most likely arises from the presence of a class of residues that play a context-dependent role in the recognition process. While R163 appears to perform a singular function in translational stalling, the other residues between 150 and 165 that play a "secondary" role probably fall into at least one other functional class. The variable contribution of these residues to translation arrest and their variable nature suggests that they work in different combinations to achieve the same end. Indeed a subset of unique residues [e.g., P160 and P161, which are the only two residues found in Sup1 but not SecM(Eco)] must specify the differences in the crosslinking patterns that we observed. Presumably the requirement for these context-dependent residues reflects the unusual circumstances of polypeptide recognition in the ribosome tunnel, an environment in which the composition and location of potential ligands is in constant flux. An alternative explanation of the plasticity of the SecM arrest motif is that both R163 and the secondary residues are coordinately recognized, but that the latter simply form less significant interactions with the ribosome than R163. This explanation is very unlikely, however, because it would imply the existence of an exceptionally complex recognition process in which components of the ribosome tunnel integrate information obtained through interactions with a potentially very large number of different sequences.
Based on the notion that the C-terminal residues of SecM perform distinct functions, we propose a new model for the recognition of the arrest peptide in the ribosome tunnel (Fig. 7). In our model, SecM residues near the top of the tunnel specify the conformation of the nascent chain and thereby influence the positioning of R163. Translation arrest occurs when the conformation of the nascent chain moves R163 into a precise intra-tunnel site where it either transmits a signal by interacting with a critical ribosomal component or simply prevents further movement of the polypeptide chain. In this model, nascent chain "discrimination" occurs relatively close to the PTC. Nascent chain conformation might be influenced at least in part by microenvironments in the tunnel. In principle, nascent chain conformation might change dynamically during polypeptide elongation and be subject to constant monitoring by the ribosome. Alternatively, a specific conformation might become locked in only when P166-tRNA occupies the A site and initiates a series of events that alter the tunnel environment or, as previously proposed, transiently stalls translation (Muto et al., 2006). Consistent with this possibility, part of the SecM(Eco) nascent chain that resides inside the tunnel undergoes a dramatic conformational change when P166-tRNA binds to the ribosome (Woolhead et al., 2006). In any case, we propose that the presence of a proline at residue 161 enhances the positioning of R163 by constraining nascent chain conformation. In contrast, when residue 161 is a flexible glycine, residues further down the tunnel such as F150 and W155 are required to stabilize the arrest peptide possibly by interacting with tunnel components. Regardless of their mode of action, F150 and W155 promote translation arrest only when compaction of the nascent chain between residues 135 and 159 moves them into an appropriate location (Woolhead et al., 2006).
An implicit postulate of this model is that the conformation of the nascent chain near the top of the tunnel is determined by multiple residues, including residues that can be located relatively far away. The prevalence of partial arrest defects that we observed is consistent with this idea because if multiple residues contribute to the positioning of R163, then a single mutation might create only a small structural perturbation. In this regard it is noteworthy that I162 is invariant in all SecM homologs, but an I162A mutation causes only partial defects in the arrest mediated by both SecM(Eco) and SecM(C-Ms). It seems likely that this amino acid plays a particularly important, yet non-essential role in the positioning of the adjacent R163 residue. The observation that the incorporation of Bpa into SecM(Eco) at positions N-terminal to the arrest motif (e.g., 140) abolishes translation arrest (Fig. S4), however, clearly suggests that distant residues can also influence events close to the PTC. Likewise, the finding that L22 mutations impair SecM(C-Ms)-mediated arrest even though the arrest motif does not reach the constriction point in the tunnel (Fig. S5) raises the possibility that the mutations act indirectly by perturbing the disposition of distant residues that normally do not participate in translational stalling. Indeed the constriction point may appear to be a universal "discriminating gate" largely because mutations in this region often exert indirect effects on the arrest process.
Finally, our results may help to explain the striking heterogeneity of arrest peptides that have been identified in both prokaryotic and eukaryotic systems. In essence, we found that the SecM arrest motif is comprised of only two essential elements plus an equal or greater number of variable elements. Consequently the SecM sequence diverges to such a degree that identification of SecM homologs would be difficult if they were divorced from their genomic context. If one postulates that all arrest peptides likewise contain only a few essential elements and that arrest peptide discrimination involves the positioning of a key residue (which is clearly not always arginine) at one of multiple sites throughout the tunnel (rather than at a fixed "gate"), then the sequence and size diversity of arrest peptides can be explained. Consistent with this hypothesis, TnaC contains only two invariant residues, an aspartate and a tryptophan located 8 and 12 residues, respectively, from the PTC. The mutation of either residue abolishes TnaC-mediated arrest (Cruz-Vera and Yanofsky, 2008). Interestingly, the residues located between the invariant residues and those C-terminal to the aspartate are loosely conserved (suggesting that they play a significant, though less critical role in arrest) whereas the N-terminal residues show no conservation. It is also noteworthy that most characterized translation arrest phenomena require the presence of a small molecule, such as an antibiotic that targets the ribosome. The binding of specific macrolides in the tunnel, which is required for translation arrest mediated by the ErmC peptide, might alter the tunnel so as to create new sites of peptide discrimination. It is also possible, however, that the binding of other small molecules such as tryptophan, which appears to bind near the A site to promote TnaC-mediated stalling (Gong and Yanofsky, 2002), directly or indirectly affects the environment in the tunnel. Indeed by binding to different locations on the ribosome small molecules might induce the formation of a multitude of discrimination sites that initiate arrest in response to only one or two critical amino acids.
CU164 (MC4100 zhd::tet secY39cs) (Baba et al., 1990), C41 (Miroux and Walker, 1996) or SQ351 (MG1655 Δ7rrn ΔlacZYA pKK3535 pTRNA67) (Cruz-Vera et al., 2005) or their derivatives were used in all experiments.
To examine the kinetics of SecM translation arrest, E. coli harboring pSecM derivatives were grown at 37°C in M9 medium containing 0.2% glycerol, 100 μg/ml ampicillin, and all the L-amino acids (40 μg/ml) except methionine and cysteine. Overnight cultures were washed and diluted into fresh medium at OD550=0.02. When the cultures reached OD550=0.2, synthesis of plasmid-borne genes was induced for 30 min by the addition of 200 μM IPTG. Pulse-chase labeling was conducted as described (Ulbrandt et al., 1997). SecM-containing polypeptides were then immunoprecipitated with rabbit polyclonal antibodies against synthetic peptides corresponding to residues 30–54 and 127–142 of SecM (Nakatogawa and Ito, 2001). Except where noted, immunoprecipitated proteins were resolved by electrophoresis on 12% NuPAGE minigels (Invitrogen) with MOPS running buffer. In these and all other experiments that involved the detection of a radioactive signal, gels were scanned using a Fuji BAS-2500 phosphorimager.
E. coli C41 S-30 extract preparation, coupled transcription-translation reactions and cetyl trimethylammonium bromide (CTABr) fractionations were performed essentially as described (Woolhead et al., 2006). In a typical experiment in which radiolabeling was required, 10 μCi Tran35S-Label (MP Biomedical) was added to a 25 μl reaction. Samples were resolved by PAGE as described (Woolhead et al., 2006) except where noted. For photocrosslining experiments S-30 extracts were prepared from strains MNY9 (C41 prfA1 zcg174::Tn10 pBpaCm) and MNY10 (C41 prfA1 zcg174::Tn10 pAzpCm) grown at 37°C in medium supplemented with chloramphenicol (10 μg/ml). RF1 was inactivated by heating the extracts at 42°C for 16 min. Reactions (200 μl) were programmed with full-length SecM variants harboring the desired amber mutation and contained 10 μCi Tran35S-Label and either 1 mM Bpa or 2 mM Azp (Bachem). After a 30 min incubation at 37°C, reactions were placed on ice for 5 min. A portion (5%) of the reaction was withdrawn for CTABr fractionation followed by immunoprecipitation with anti-SecM. The RNC complexes in the remaining portion were isolated by centrifugation through a 0.5 M sucrose cushion and resuspended in 200 μl RNC buffer (Woolhead et al., 2006). A portion (50 μl) representing untreated RNC complexes was TCA precipitated and subjected to immunoprecipitation with anti-SecM. The remainder was transferred to a 96-well flat bottom plate and UV-irradiated (365 nm, Spectroline SB-100P) on ice for 15 min at a distance of 3–4 cm. A portion (50 μl) of the irradiated RNC complexes was treated as described above and represented the UV-treated samples. The remaining ~90 μl were TCA precipitated and subjected to immunoprecipiation using rabbit antisera raised against synthetic peptides corresponding to the N-terminus of L4, L22 and L23 (MELVLKDAQSALTVSETTFGC, METIAKHRHARSSAQKVRLVC, MIREERLLKVLRAPHVSEKAC). Finally, all samples were resolved on 12% NuPAGE minigels with MOPS running buffer.
We thank John Atkins, Lauren Bakaletz, Alexander Mankin, Greg Phillips, Peter Schultz, Cathy Squires and Michael Yarmolinsky for gifts of plasmids and strains and Will Prinz and Gigi Storz for critical reading of the manuscript. This work was supported by the NIDDK Intramural Research program.
Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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.


text:
Knowledge and experience, along with leading-edge technology, and a commitment to integrity, give D&M the ability to provide the highest levels of service possible to its clients.
D&M's service capacity is enhanced by the regional knowledge that is resident in each division and by the local presence of team members around the world and D&M's willingness to go where customers need us.
For many of D&M's assignments, time is of the essence, and our commitment to meet or exceed client deadlines is as critical to our evaluation as the report itself.
D&M has many long-standing relationships with clients, and many of them rely on the firm for recurring services, such as income projections based on current economic conditions or annual reviews of reserves estimates. Such enduring client relationships, allow D&M to work in a manner similar to auditing, but all D&M analyses and reports are based on sufficient independent work such that D&M can unreservedly support and defend its opinions as realistic and unbiased.

subset: commoncrawl


text:
Howard Nutt is Executive Director of the Business Development Institute International (BD-Institute). He leads operation of this international non-profit organization, which is dedicated to extending BD best practices world-wide. Howard managed creation of the BD-CMM in 2001 and has lead development of subsequent versions. He has also established a range of products and services to support adopters, assess their capabilities, and assist them in implementing BD-CMM.
Track: Professional Disciplines: Best Practices
Presentation Title: Optimizing Solutions by Integrating CMMI and BD-CMM – Exploiting the Power of a Fully Integrated Life Cycle Management Strategy
Professional Discipline: Program and Project Management
Presentation Summary: Project failures within even high-maturity organizations can often be attributed to short-comings that trace to poor pre-contract solution architectures. While organizations use CMMI to develop solutions that are more predictable and robust for customers, such application of best practice can experience limited success when solutions are fundamentally flawed during project conceptualization. The Capability Maturity Model for Business Development (BD-CMM) applies best practices for pre-contract solution development that are fully aligned with CMMI and address both contractor and customer risk. Resulting integration between BD-CMM and CMMI creates a unified strategy for managing success from end to end of business life cycles.


text:
These gadgets make gardening easier, regional experts say
By Dan Vierria, Special to The Bee, 01/01/2015
Read more here: http://www.sacbee.com/entertainment/living/home-garden/article5282964.html#storylink=cpy
Tool: Bulb auger
A self-described "gadget geek," Pratt had difficulty choosing just one tool. After much deliberation, she chose the bulb auger, a long, metal drill attachment used to dig holes for bulbs and other plants. Pratt paid "around $25" for the auger that has a 2.5-inch diameter and is 24 inches long.
"It makes quick work of bulb planting and turned a formerly dreaded task into something I now enjoy," Pratt said. "It's an energy-saver, a time-saver and a back-saver. Mine has a long shank and spiral cutting blade. Look for one that also has a soil whip at the top."
She tried those "as seen on TV" hand-held, cylindrical bulb planters, but they didn't do the job.
"They were no match for clay soil," she said. "I suppose you could use them in fluffy, raised beds or in the kitchen as a biscuit cutter. In the garden? Not so much. The drill-powered auger is especially great for those of us who have to contend with clayey soils and garden beds dominated by mature tree roots."
She uses a 12V lithium ion DeWalt cordless drill to power the auger, but any comparable drill will work. The auger can be found on www.buildingsupplyplus.com or check with Amazon.com, Peaceful Valley Farm Supply in Grass Valley and Ace Hardware as possible sources, Pratt advised.
"You can use it for planting bedding plants, too," she added.
Read more here: http://www.sacbee.com/entertainment/living/home-garden/article5282964.html#storylink=cpy
Sunset Magazine
Sunset's Blog Roll
"Botanical zeal infuses these dispatches from a mom-gardener-photographer near Sacramento."
Sacramento Bee, May 2006
Best Area Blog and Site
Angela Pratt, a horticulturist, has designed a masterpiece. For area gardeners, this is a must-bookmark Web site. Her blog (personal gardening diary) is fun to read and filled with digital images to illustrate what she's planted and where she's visited. Colorful images are showcased throughout the site. Links to Sacramento-area gardening information will lead you to just about anything you desire. There's advice for planting and maintenance…. there's now a list of Northern California gardening blogs. – Dan Vierria, Sacramento Bee Garden Columnist


text:
Results 1 to 9 of 9
August 31st, 2003, 11:37 PM #1
- Join Date
- January 18th, 2005
- Posts
- 8
Hi all,
As said in my intro. I started with AM 4 mts. back and about a month back Linkshare mailed me first 2 commi$$ion checks, which I haven't received yet.
I receive other checks from some individual merchants within 1-2 weeks.
Can anyone please let me know if this is normal with LS, and what should be the course of action if the checks are misplaced or not received at all. Thanks in advance.
September 1st, 2003, 01:14 AM #2
- Join Date
- January 18th, 2005
- Posts
- 3,219
Not sure if Linkshare is your hope for a source of income or not. But for me it's ... Hey cool a check, hum who is this from. Ohhh Cool Linkshare merchant. Awesome beer money!
And that's the way I have dealt with them for 5 years!
September 1st, 2003, 01:30 AM #3
- Join Date
- January 18th, 2005
- Posts
- 2,419
If linkshare said they mailed you the checks a month ago and you haven't received them - something is wrong. I suggest you inquire specifically with Linkshare as to the status of your checks. This scenario is not a normal
Welcome to ABW.
PeePee merchants with PooPoo policies allow our earnings to be flushed down the crapper.
Why give parasites unlimited cookie durations and credit for sales where they divert our users and overwrite our cookies. PP merchants directly support what many consider unfair trade practices and thievery!
September 1st, 2003, 06:31 AM #4
- Join Date
- January 18th, 2005
- Posts
- 804
Linkshare has been sending me checks within a week of receiving funds from the merchant(s) consistently since I began with them. Any holdups I've noticed have been the merchant taking their time paying Linkshare.
When you login to your Linkshare account, click the "Payment Report" link and you can see the status of previous months. Last month's report usually get posted within a week of the end of the month, so you'll be able to see August status by about the 5th to 7th of Sept.
September 1st, 2003, 08:15 AM #5
- Join Date
- January 18th, 2005
- Posts
- 8
Thanks for that warm welcome & suggestions!
Well I am looking at AM as a serious source of income, which seems to be working in last 3 months. But as said I am facing problems with my very 1st commission checks.
I did tried contacting LS at email@example.com but heard NO REPLY. SO then I contacted Sarah Johnson at email@example.com (which I got after reading this forum), she just replied generally about getting back to me once she hears from their Accounts dept., but neither turned back.
Does anyone here knows contact mail-ID for Accounts dept at LS.
Thanks in advance.
September 1st, 2003, 11:08 AM #6
I would call them, rather than email...and yes, you can make a sole income on LS, or any other network. It all starts wth a check or two. :0)
I got my T-Shirt! Imagination that!
September 1st, 2003, 02:09 PM #7
Just a side not here.
Linkshare merchants do the best for me on a consistant basis. My worst is CJ.
Linkshare always sends out payment promptly after receiving it from the merchant in my experience.
"All I ask is a chance to prove that money can't make me happy."
"Work to become, not to acquire." -- Confucius
September 11th, 2003, 06:32 AM #8
- Join Date
- January 18th, 2005
- Posts
- 8
Hello All,
Finally, yesterday I got my checks. They were reissued by Linkshare. Thanks to Linkshare and this Forum for being supportive and advisable.
September 11th, 2003, 06:43 AM #9
- Join Date
- January 18th, 2005
- Posts
- 8
Well, There's few more thing...
The envelop in which I got my checks (finally) had my address HAND WRITTEN (not TYPED) on it.
Second the address format was as...
...Postal Address...
(My name was not mentioned on the envelop)
Is this how you guys get your checks too?
Please let me know. Thanks in advance.
Similar Threads
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Missing Linkshare checksBy Grumpy in forum Rakuten LinkShare - LSReplies: 7Last Post: October 12th, 2003, 11:47 AM
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text:
Shirley Jackson's short story, "The Lottery," appeared in the June 26, 1948, issue of The New Yorker. The response to the fictional piece, which concerns a small-town tradition that ends in horrific violence, was profoundly negative. While many were dumbfounded by Jackson's story, others were deeply disturbed. Here's one such response: "I read it while soaking in the tub…and was tempted to put my head underwater and end it all." Readers were shaken and outraged by it—even Jackson's own parents let her know they didn't like the piece. In all, the author received over 300 letters about "The Lottery"; only thirteen were positive. But what was the moral of the story? Thoughtless conformity can lead to cold-blooded killing? Ordinary people are capable of committing unspeakable atrocities? Cruelty is random? In July 1948, Jackson explained what she was trying to convey.
I suppose I hoped, by setting a particularly brutal ancient rite in the present and in my own village, to shock the story's readers with a graphic dramatization of the pointless violence and general inhumanity in their own lives.
Two decades later, a film adaption of "The Lottery" was produced by the Encyclopedia Britannica Educational Corporation. The 1969 short went on to become one of the bestselling educational films of all time.
I first saw The Lottery in a middle school English class, and was around twelve years old. I was wholly unfamiliar with Shirley Jackson's story, and as my teacher didn't introduce the film, I was completely unprepared for what I was about to see. During the final moments, the matter of fact nature of the violence was so shocking…I mean, I was stunned. STUNNED. When the film ended, so did class, and I have no memory of talking to anyone about the film. What I do remember is that I had gym next, and vividly recall sitting on a bench in the boy's locker room, dazed and shell-shocked, trying to make sense of it all. The Lottery scared the shit out of me, and I've never forgotten that moment. Thanks to the Internet, I know now I wasn't the only one so strongly affected by it.
I never read the short story, so watching this short film was a true shocker for me. Like many other people, I saw this in my English class a long time ago, and since then, I still haven't seen it. But I still remember that time; it really stays with you. I remember everyone in my class with their jaws dropping, we couldn't believe it. (IMDb user review)
A young Ed Begley Jr. appears in the film, as does the prolific, but lesser known actor, William Fawcett, who steals the show as "Old Man Warner," the curmudgeonly elder whose blind faith in the lottery tradition is ripe for analysis.
In addition to the film, the Encyclopedia Britannica Educational Corporation also produced a nine minute examination of The Lottery, and it's totally worth watching (it immediately follows the eighteen minute short, embedded below). I wish my English teacher would've made time for that, too! If, like the twelve-year-old me, you've never read Shirley Jackson's controversial allegory, do yourself a favor and feast your eyes on it here. A fine collection of the author's novels and short stories, which, of course, includes "The Lottery," is for sale on Amazon.
Previously on Dangerous Minds:
'The Finishing Line': The grisly British educational film that scared kids and shocked parents


text:
Install Whonix in Qubes
Installing Whonix in Qubes is simple and only requires a few simple steps.
First Time Users
Using privacy and anonymization tools like Whonix is not a magical solution that instantly makes you anonymous online. Please consider:
- Do you know what metadata or a man-in-the-middle attack is?
- Do you think nobody can eavesdrop on your communications because you are using Tor?
- Do you know how Whonix works?
If you answered no, have a look at the about, warning and do not pages (on the Whonix website) to make sure Whonix is the right tool for you and you understand it's limitations.
Install Templates
Launch the
dom0 terminal
Konsole from your Qubes App Launcher. Then enter the following command to install the Whonix-Gateway and Workstation TemplateVMs.
sudo qubes-dom0-update --enablerepo=qubes-templates-community qubes-template-whonix-gw qubes-template-whonix-ws
Download will take a while and there will be no progress indicator.
After doing this, you should see two new TemplateVMs in the VM Manager called
whonix-gw and
Enabling AppArmor
This is an optional security enhancement (for testers-only). If you're technical & interested, see Enabling AppArmor.
Configuring Whonix VMs
Create a Whonix-Gateway ProxyVM by clicking on
Create a New VM and select
whonix-gw as the template and select ProxyVM as the type.
Create a Whonix-Workstation AppVM by clicking on
Create a New VM and select
whonix-ws as the template and select AppVM (should be default) as the type.
Configure the Whonix-Gateway TemplateVM to use the
sys-whonix ProxyVM that you just created.
Great. You should be all done installing Whonix into Qubes. Use these two TemplateVMs and the ProxyVM you just added, like you would for any other VMs.
Running Applications
To start an application in the Whonix-Workstation AppVM that you created, launch it like any other- open the
Qubes App Launcher and then select an app such as
Privacy Browser which will then launch the Tor Browser
Advanced Information
You can learn more about installing Whonix, Qubes-Whonix or Whonix generally.


text: I stumped everyone with my App2SD question, I hope I don't do so again with this one. I bought a wifi Kindle which will not accept peer-to-peer or enterprise connections. I tried to use my DX as a wifi hotspot using Barnacle and Wireless Tether For Root but unfortunately they seem to only be peer-to-peer and not the "access point" I need. Is there an app which will turn my phone into an access point wifi hotspot? I'm not well versed on wifi so I don't understand if there is some hardware limitation to doing this or not.


text:
Bill and Barbara Walsh, who spearheaded fundraising efforts at Philadelphia Country Club for many years, were honored with the first J. Wood Platt Distinguished Service Award at the 2012 Caddie-Scholar Brunch. As the first recipients of the award, it will named in their honor for perpetuity. Philadelphia Country Club has been the top fundraising GAP Member Club for the Platt the last nine years, topping $100,000 in yearly contributions eight times.
Sadly, Mr. Walsh passed away December 18, 2013. The below story speaks to the impact he had on others
By Fred Behringer
The Golf Association of Philadelphia lost one of its most vigorous advocates and outstanding volunteer leaders with the death of William T. Walsh, 91, of Devon, Pa., on Dec. 18.
Beginning in the 1950s, Mr. Walsh competed frequently and successfully in GAP championships and Team Matches. He served for a record 13 years on the GAP Executive Committee, including three years as president, and he is credited with spearheading the decision in 1996 to admit public golf clubs to membership in GAP.
He also played a prominent role in raising funds for GAPs J. Wood Platt Caddie Scholarship Trust. With Mr. Walsh as its Platt chairman for 10 years, Philadelphia Country Club led all GAP clubs in scholarship donations in nine of those years, and it holds the record for total contributions by a club.
In 2008, GAP awarded Mr. Walsh its Distinguished Service Award for exemplifying the spirit of golf at its highest level and making a substantial contribution to the game. By word and deed, he promoted golfs traditions and ethics, volunteering immeasurable hours to advance GAPs programs.
Recalling that it was Mr. Walsh who encouraged him to get involved in amateur golf administration, Frank E. Rutan, IV, president of the GAP Executive Committee, said, We will be forever grateful for all the work he did on behalf of the Golf Association of Philadelphia and the J.Wood Platt Scholarship Trust.
Clark ODonoghue observed Mr. Walshs commitment to caddie scholarships as chairman of the Platt Trust for several years. Bill Walsh was legendary on so many different fronts, ODonoghue said. He was such a force not only with the fundraising at Philly Country Club, but he put a charge out there to the rest of GAP. The way he got Philly out there as the leader really spurred on other clubs to boost their efforts. He was in the forefront of that.
Stan Engle, a past president of Philadelphia Country Club and a close friend of Mr. Walsh since the 1970s, said, Bills the kind of guy who, from a golf perspective, I would never think about whether or not he would call an infraction upon himself or cause an infraction. He did the right thing in his life at all times, and he taught others through his examples. He was wond
Craig Ammerman, a past president of GAP who also served on the United States Golf Association Executive Committee, felt that Mr. Walshs quiet diplomacy in persuading member clubs to support the admission of public golf clubs to membership was his most important contribution to GAP and that this vote ranks as GAPs most important act. Mr. Walsh, he said, was likely the only person who could have made this happen.
Since welcoming public clubs, Ammerman pointed out, GAP has grown from 93 member clubs to 184. This is staggering growth when you consider going through years when the economy was not very favorable to golf. It couldnt have happened if we had not opened our arms to places where people play golf.
Ammerman also credited Mr. Walsh for really invigorating Platt Trust fund-raising. You wont find a better gentleman in the world than Bill Walsh, he said.
Mr. Walsh began playing golf at age 10 in New Jersey, caddied at Metuchen Golf & Country Club and won two championships there and one at Plainfield Country Club before moving to the Philadelphia area in 1955. He later added two club championships at Skytop Golf Course and seven at Philadelphia Country Club. He shared 10 GAP Father & Son championships with four different sons. He considered his 1991 GAP Senior Amateur Championship his most significant golf title.
A longtime member of Pine Valley Golf Club, Mr. Walsh sponsored the Walsh Open there in November for 50 years, beginning in 1958. The invitational outing for his friends began with one foursome and grew to 40 golfers each year. Mr. Walsh typically would risk $2 wagers in such friendly competition, and he collected most of the time. Conversations at times turned slightly acerbic. While a friend once said, Bill has the sharpest needle of all, Mr. Walsh could take the jabs as well with self-deprecating humor.
Mr. Walsh was a past president of the Philadelphia Seniors Golf Association, an organization of members of GAP clubs founded in 1920. He posted the low gross score 49 times in the groups outings.
He also belonged to Jupiter Hills Golf Club and Tequesta Country Club in Florida.
Another aspect of Mr.Walshs love affair with golf involved keeping meticulous track of the courses he visited. A round at Augusta National in 2002 became his 500th different course.
He also counted the number of times he shot his age or lower. He did it for the first time at age 72 and exceeded it 300 times by the end of 2011.
Mr. Walsh worked all his golf activities into a schedule that included managing a business and serving as a leader in community, religious and professional organizations. He once estimated his daily attendance at Mass through 2011 totaled more than 23,500 visits.
Mr. Walsh was born in 1922 in Westfield, N.J. He graduated from the Villanova Commerce and Finance School and played on the golf and basketball teams. After serving as an officer in the Navy during World War II, he graduated from the University of Pennsylvania Law School. He practiced law only briefly before entering the insurance business with the Equitable Life Assurance Society in New Jersey. He came to Philadelphia for a promotion to agency manager, eventually establishing Walsh Associates in Wayne, Pa., currently operated by three of his children. Among many business commendations were his selection as Equitables National Honor Agent and membership in the companys Hall of Fame.
Mr. Walsh maintained a close relationship with Villanova throughout his life, serving as president of the General Alumni Association and as a member of the University Development Council. He received the Villanova Loyalty Award and the Villanova Alumni Award.
As a longtime, heavily involved member of the Rotary Club of Ardmore, Pa., Mr. Walsh served as club president and as district governor for Southeastern Pennsylvania. He completed more than 40 years of perfect attendance.
Mr. Walsh is survived by his wife of 65 years, Barbara Straub Walsh, their 15 children, 29 grandchildren and 10 great grandchildren.
One of Mr. Walshs sons, Paul Chet Walsh, a leading amateur golfer in the Philadelphia region, said of his father, Whenever he got into anything, he really put everything into it, and this was especially true of the Golf Association. He really knew how to talk to people. The amount he raised for the Platt Trust at Philadelphia Country Club has been just amazing. When you mentioned something, he was already walking the walk. He never tackled anything without getting fully involved."


text:
105 Maple Street Cooroy 4563 QLD
cottages like this are hard to find
This is a well maintained one level timber home on town block featuring hardwood floors throughout, 3 good sized bedrooms, a renovated bathroom with a spacious kitchen and living area with ceiling fan.
The block is very private and fully landscaped for gardeners, with plenty of room for hobbies & pursuits.
The property is located close to absolutely everything the historic town of Cooroy has to offer.
A short walk to modern shopping, medical facilities and private & state schools or catch the tilt train from Cooroy station.
Easy access for the disabled is in place and the house has been renovated inside and out so virtually nothing to do but move in and enjoy the home with under cover car parking with town water & sewerage.
Please call or email for further information on this property.
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Cooroy QLD, 4563
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text:
I have several very old pieces of cast iron. Each one has an almost mirror like finish on the surface and I know that they are well seasoned and sometimes food still sticks. I've looked at new CI in stores and I've noticed that all of them have a very rough, sandlike surface. I assume that this surface is imparted to the metal through sand casting, the process of molding compressed blocks of sand and pouring molten iron into the cavity.
Seems to me that this surface is not conducive to any non-stick cooking no matter how well seasoned the utensil is.
Anyone else have any ideas on this.


text:
VideoHive Drawing Heart 8879041
Stock Footage \ Lifestyle
Valentine \ Pencil \ Contains \ Version \ Sketch \ Marriage \ Drawing \ Couple
Stock Footage - Drawing Heart 8879041 by limette @ VideoHive
Stock Footage \ Lifestyle
Drawing heart with pencil. You can use for show your love. Contains two version.
Alpha Channel: No; Looped Video: No; Frame Rate: 25; Resolution: 1920x1080; Video Encoding: H.264; File Size: 512mb; Number of Clips: 2; Total Clip(s) Length: 1:24.
Smartphone; City Map Guide; Billboard; Mobile Phone; Hands; Mobile Phone; Smart Phone.
Keywords: couple, drawing, heart, love, marriage, pencil, sketch, valentine, valentine's day.


text: I wanted to share a very useful app I found recently. I recently picked up two 16GB HTC Flyers. They come with about 8GB of free internal sdcard space. The external card is seen as /sdcard2 or /sdcard/ext_sd so I found the huge gameloft games and others like deadspace, GOF 2, NFS Hot Pursuit, etc were quickly filling up my 8GB on /sdcard. I did some searching on this problem and found directorybind. I was able to move the largest game data directories from /sdcard/Android/data/.... to /sdcard2/Android/data/... and they are working flawlessly. The app will move the files for you and will bind the directories on reboots too. FYI I had to tick the alternate database option. The app can be found here:


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Logitech Z520 Review and Ratings
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Our Verdict: This two-piece speaker set delivers airy sound and cut-above audio quality for basic desktop-bound music listening and movie watching, but we'd look for them on sale. Read More…
- What We Liked…
- High-quality, spacious sound
- Reasonable bass without a subwoofer
- What We Didn't…
- Pricey for a stereo-only set
- Hardwired cabling
Logitech Z520 Review
By September 21, 2009reviewed
Here in the late '00s, the well-dressed desktop-PC entertainment station isn't au courant without an elaborate multichannel speaker system hanging off its audio ports. So, in this age of eight-channel surround audio, positional sound for games, and mind-boggling tangles of cable, it's refreshing to see an old-school speaker set that harkens back to a simpler time. Logitech's Z520 is a bulky, weighty, two-piece PC-speaker set that delivers straight-up, no-nonsense desktop sound. There's no subwoofer, no software-processing extras, and just one control knob. There is, however, a big price tag: $129.99 direct from Logitech. (At the time of this review, it was available for $10 to $20 less from various resellers.) The Z520s delivered above-average stereo sound in our tests, but their maximum volume levels, basic design, and lack of convenience features make us wish they cost a third or so less.
The Z520 set is the top-end speaker set of four recently introduced by Logitech. (The Z523, at $99.99 list, is a subwoofer-equipped three-piece ensemble, with the Z320 and Z323, both lower-end at $69.99, being step-down models.) We were intrigued that a premium two-speaker set, rather than a three-piece one, would top the line, and set the speakers on our test bench to hear them out.
Setup was trivial: Connect a hardwired cable (with dual RCA jacks at one end) from the left speaker to the back of the right, then position the speakers on your desk. A single miniplug running from the right speaker goes into your sound card's or motherboard's front-speaker output (usually a jack color-coded green), and you're done. All that's left is to go into Control Panel (assuming you're running Windows of some stripe) and/or your sound card's or chipset's software, to check that your audio configuration is set up for stereo speakers. One quibble—although speakers get little wear and tear, we think that for a premium-priced set like this one, the cabling shouldn't be hardwired into the speakers; detachable cabling can be replaced if it goes bad.
The speaker bodies take their design cues from computer speakers, not home theater. Standing just over 9 inches high, they're tapered in the back, with a weighty base. Indeed, they look a bit old-fashioned for 2009-model computer speakers— "old-fashioned" in this context meaning "last-decade." Although their sound would work well for, say, a small bedroom's A/V arrangement, they look like they belong on a desk.
Controls (or rather, "control") couldn't be simpler; a single knob on the face of the right speaker governs volume and on/off. You don't get bass or treble controls. The only other sops to a feature list are microphone and headphone jacks on the right side of the right speaker; use of the headphone jack auto-disables the speaker output.
The speaker bodies have removable faces that protect the actual cones. Each satellite has two cones: a 1-inch tweeter (which also vents to the rear, ostensibly contributing to Logitech's claim of "360-degree sound" for these units) and a 2-inch combined midrange/woofer. The grilles are rigid and durable perforated metal, well able to protect the domes.
We cranked up the Z520s with a variety of audio content: music from CDs and MP3s (quiet and bombastic classical, harsh electronica, jangling pop-rock), an action movie (a recent James Bond title), and several games (including Half-Life 2, an explosion and gunfire-heavy shooter, and Bejeweled, with its spacey, ambient soundtrack). For the purposes of our tests, our sound card, a Creative Sound Blaster Audigy Platinum, was configured for stereo two-speaker output, and the system-level bass/treble controls were set at 0dB, with no audio processing. All system volume controls were set at 85 percent, to avoid top-end distortion by overdriving the sound card.
For a two-piece set, the Z520s delivered surprisingly airy, wide-sounding audio. The music, in all cases, sounded brilliant, though the bass was not well-defined. We were especially impressed with our classical trial (Moussorgsky's Pictures at an Exhibition), which well-showcased the breadth of the sound. Interestingly, the compressed nature of our test MP3 files, when played in an A-to-B comparison with their CD originals, was clearly audible on the Z520, especially in high tones. With many PC speakers, it's hard to tell the difference between an original CD and an MP3 file, but not with these.
Movie playback was even better. The spatial feel of Quantum of Solace's dialog, music, and audio effects surpassed what we've heard from most two-piece PC-speaker sets we've tested. While the sound wasn't strictly positional, we noted a clear sense of airiness, separation, and space. We compared these to our test-bed PC's Altec Lansing 251, a six-piece set that was well-regarded about three or four years ago (and was priced roughly the same). The Z520s' sound was markedly less muffled-sounding and airier. (Admittedly, the Altec Lansing's satellites are much smaller.)
In our gaming trials, we noted no clipping or squelching of transient highs like gunfire, even with the volume turned way up. By nature, though, serious gamers probably won't find these speakers a good fit; you can buy five-piece sets that allow for true positional audio in games for the same price or less. An infrequent gamer, however, whose greater need is audio precision primarily for music will be happy with the Z520.
Across all audio types, though, we noted a lack of truly room-shaking volume. As noted, we set all system-level volume controls (and any relevant application controls) at 85 percent, to avoid top-end distortion at the sound-card level, and we cranked the Z520s all the way up. The speakers certainly delivered sound that could be heard across a large room (especially with our rock-music samples), but the lack of a subwoofer was evident. If you tend to push your speakers hard (and your neighbors don't complain), you would probably prefer a set with more muscle. At top volume, the impact of the sound faded away fast much beyond 10 feet away; indeed, the audio was clear but didn't pack much punch from that distance. Expect to get the best results from these speakers when you're parked right in front of them at your desk.
Speakers are, of course, all about personal preference, so we suggest auditioning this model and others in its price range in person before buying, since the Z520s are not quite an impulse buy. But if you can get them marked significantly down from the $129.99 direct price, you'll find them a super-solid pick for enjoying music and movies from your desktop, without the cable complexity of a 5.1 set or the clutter of a subwoofer.
Logitech Z520
Our Verdict: This two-piece speaker set delivers airy sound and cut-above audio quality for basic desktop-bound music listening and movie watching, but we'd look for them on sale.
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Key Specs
70Hz to 20KHz
9.1x4.8x6.5 inches (each satellite)
26 watts
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i think what people forget is that in terms of gameplay ico wasnt that special. block puzzles platforming and tricky controls. likewise shadow got 8s across the board and could be infuriating at times. in shadow if your grab meter ran out at the wrong time down you went.
while the gameplay is not that special what this and the other games in the series do have in buckets is atmosphere and fantastic world building. not many games can draw me in and make me stop and go wow at there sense of scale. i really like the feeling that i'm one of the first people to explore these ancient places in 100s of years.
Like the horse in shadow, or yorda in ico secondry charicters have there own personality and own free will, i like the fact that controling trico isnt like controling a big tank hes got a real personality.


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2008-20-27 - Eurocopter 2500 USAAmerican Eurocopter celebrates 2500th U.S. delivery
2500 Eurocopter-Hubschrauber in USA geliefert<br /> At an event on its booth at the Air Medical Transportation Conference in Minneapolis, MN, American Eurocopter celebrated its 2500th U.S. helicopter delivery with long-time customer PHI, Inc., one of the world's leading helicopter services companies.
The 2500th helicopter is an EC135 that is scheduled to be delivered to the PHI Air Medical Group shortly after AMTC. "This is a momentous occasion for us and we are delighted to share it with our valued friends at PHI," said Marc Paganini, President and CEO of American Eurocopter. "This is a great testament to the history of our company and this achievement reflects American Eurocopter's heritage and its development in the United States." "The delivery of this EC135 to PHI is especially significant, since the very first helicopter delivered into the United States was an SA.3130 Alouette delivered to PHI in 1957," explains Paganini. Since that time, PHI has operated a wide variety of Eurocopter aircraft and PHI's fleet today includes AS350B2/B3 AStars, EC135s and BK117s.


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Notes from Portland, Seattle, Minneapolis, and New York. That's more than enough travel for one month.
Semifat Sediment is Josh Lee's weblog. Since 2003, Josh has been using this site to record his thoughts on games, television, reading, writing, and whatever else happens to be occupying his attention at the moment.
And don't try to make sense of the site's name; it's just two words that Josh liked the sound of when he was first setting up the site.
More about Semifat Sediment »
Like everyone else these days, Josh is scattered all over the Internet. Here are a few of the places you can go to try and put a picture of him together:
Semifat Sediment © 2003-2012, Joshua Lee. All rights reserved.


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Marketmore 76 Slicing Cucumber - Cucumis Sativus - 20 Seeds
Seeds for Africa
Long, slender, dark green cucumbers. The slender, refined "Marketmore " has 8-9" fruits staying uniformly dark green even under weather stress. Begins bearing late, but picks for a relatively long time. Intermediate resistance to cucumber mosaic virus, downy mildew, and powdery mildew.


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UK legal aid cuts 'may spell the end of multinational accountability overseas'December 2011
A law firm has warned that the UK government's proposed changes to legal aid could make claims against multinationals for human rights abuses committed abroad "all but impossible".
The human rights lawyers Leigh Day said in an interview with the Guardian newspaper that a new bill making its way through the House of Lords would effectively prohibit all but the most straightforward legal cases in an area where the issues are rarely clear-cut.
The parts of the Legal Aid, Sentencing & Punishment of Offenders Bill that refer to "litigation funding and costs" will require claimants to meet the cost of "success fees", limiting the scope for complex cases to be pursued on a no-win no-fee basis where, currently, claimants are able to force guilty defendants to pay their legal fees.
There are also concerns that the introduction of a "proportionality" clause on litigation costs will essentially exclude human rights claims, which often cost more to pursue than the compensation sought.
Leigh Day said: "The effects would be to drastically curb the availability of access to justice by individuals whose human rights are abused by British companies in their overseas operations."
The Ministry of Justice noted in response that "the costs involved will be more proportionate to the sums in issue".
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Leigh Day | UK & NI Ireland | Corporate LawFurther Information


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Kenya: robbers kill priest
A Kenyan priest died in a Nairobi hospital of gunshot wounds on Friday, a week after being attacked by robbers. Fr Martin Macharia Njoroge, was travelling in his car when some bandits pulled him over in the outskirts of the Kenyan capital. "After dragging him from his car they shot him four times at close range", the Bishop of Nairobi, Archbishop Raphael Ndingi Mwana'a Nzeki told the Missionary News Service. "He did not resist them, but apparently even handed over the keys to his car, which the bandits left 500 metres away. His body was left on the ground, then found by a 'good Samaritan' who took him to the hospital". Fr Martin arrived at Nairobi's Agha Khan Hospital with two bullets in his lungs, one in his right leg and another in his chin. After a few days three bullets dislodged, while a fourth, in his left lung, had to be removed surgically, but Fr Martin died on the operating table. "Padre Martin died from a cardio-circulatory arrest during the operation", the Archbishop said. Fr Martin was ordained priest only four years ago and served at the St Francis Xaverian parish of Parklands, in Nairobi. His death is another terrible loss for his family and the diocese. Six priests have met violent deaths in the archdiocese of Nairobi in recent years. The Kenyan government of Mwai Kibaki has just opened an inquiry into the mysterious shooting of Fr John Anthony Kaiser, missionary of the St Joseph Mill Hill Society, who was found dead 24 August 2000 near Lake Naivasha. Fr John was known for his campaigning and work for the poor.


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Vintage Mamod Stationary Steam Engine In Very Good Condition
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- Sold Date:
- Source: eBay UK
Vintage Mamod stationary steam engine. It appears that it has never been fired up and all the solders/unions are in good order and moving parts all work so it should work if you wanted it to. It is missing the burner tray but overall condition is very good. Any questions, please ask.
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Wedding dresses, really? You might as well get married in voiles or naked altogether.
Warren Beatty: actor, director, writer, producer.
***** celeb
those are some long necks.
I'm open to everything. When you start to criticise the times you live in, your time is over. - Karl Lagerfeld
Had I been at this show. I would have been fighting the urge to throw sandwiches at these brides, instead of rice.
You don't engage with crazies. Because they're, you know, fucking crazy. - WitchCurlGirl
When I was shopping for a gown, I tried on several of hers. They were magnificent. The fabrics were just stunning. She's taken a 90 turn since then. Some of these would be pretty if modified to be a bit more modest but the rest are ridiculous.
Yes, some of the skirts are glorious but the tops are too sheer.
I have some famous friends and I have mostly not famous friends.
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These items are not for sale and the descriptions, images and prices are for reference purposes only.
Scissor chair in the style of Pierre Jeanneret, yellow vinyl with black trim, Australia, c 1950s
Pierre Jeanneret (Swiss, 1896-1967), Lounge Chair, designed c. 1955, teak frame, seat with braided cane work, height 75.5 cm width 65 cm depth 48.5 cm. Provenance: Cf. for comparison a similar chair in the collection of the City Museum of Chandigarh
Le Corbusier, Pierre Jeanneret and Charlotte Perriand, LC4 chaise longue, with polished trivalent chrome plated steel frame and black leather upholstery and head rest. Provenance: This example was purchased from the Conrad store in London
Pierre Jeanneret (Swiss, 1896-1967). Conference chair, designed circa 1952 teak frame, with can seat
Pierre Jeanneret (Swiss, 1896-1967). Conference chair, designed circa 1952 teak frame, with can seat, painted inventory numbers
Pair of LC1 lounge chairs Ny Le Courbusier, Pierre Jeanneret & Charlotte Pierrand for Cassina, Italy, black leather and chrome, designed 1929 stamped 'Cassina' and numbered 'Lc1 24174 / Lc1 24146'
Charlotte Perriand (1903-1999), Le Corbusier (1887-1965), Pierre Jeanneret (1896-1967), a Lc-4 chaise longue, designed 1928, manufactured by Cassina, Italy, c.1985 chromed metal, steel, cow hide, leather stamped with number, 32 PN6 = Pe Bo - 32 - 312 Uni 7
Le Corbusier, Pierre Jeanneret and Charlotte Perriand, a pair of Basculant chairs, model B301, after the original 1928 design, manufactured by Cassina. Chromed bent tubular steel with black leather seats, backs and sling arms.
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CityParks Theater: National Theatre of the USA's Chautaqua
along the East River btwn Grand & Jackson Streets
The NTUSA will channel the form and style of the original Chautauqua Lectures, a wildly popular lecture circuit that flourished from 1874 to the Great Depression, through its own inimitable aesthetic and theatrical rhythm, exploring the still hotly debated relationship between High Culture and the Mass Mind. Chautauqua! combines the history of American popular art and entertainment, the culture of commerce and the commerce of culture - our intellectual and political heritage.
Click Here for more info


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Rishita Manhattan – ...
₹ 4,000,000.00
₹ 20,000,000.00
Nimai Familia is a new modern residential project by Nimai Group located at Sector – 7, Sohna Road, Gurgaon. This Project offers 2 and 3 bhk apartments with lots of amenities. It is as well sided by a 50 metre green girdle on moreover side, building it a peaceful and wonderful living legroom for your relatives. A swarm of up to date facilities such as the Retail Outlet, a fashionable club by way of swimming pool; steam as well as sauna services; tennis along with squash courts, along with other leisure furthermore fitness conveniences increase your recreation in addition to expediency. For more info visit http://www.nimaifamilia.net.in/
Published date:
13/03/2015 3:34 pm
Expiration date:
Sector 7 Sohna Road, Gurgaon
|For sale
|New construction
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Rishita Manhattan – ...
₹ 4,000,000.00
Omaxe Residency – 2B...
₹ 12,000.00
Gurgaon Gateway – As...
₹ 18,000,000.00
TATA La Vida – Spaci...
₹ 16,600,000.00


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- Samsung Mass-Produces 1Gb DDR2 80nm Memory
Samsung Electronics, the world leader in advanced memory technology, announced today mass production of 1Gigabit (Gb) DDR2 DRAM memory using 80 nanometer (nm) process technology. While monolithic 1Gb DDR2 is available today, it is produced with more costly and less efficient 90nm technology.
- Agilent Acquires Xpedion – RFIC Simulation and Verification Software
Agilent Technologies today announced it has completed the acquisition of Xpedion Design Systems, a privately held company that provides software for wireless and high-speed digital circuit and systems design in the communications industry. Financial details were not disclosed.
- Earthrace Powerboat Heads Around the World with Xilinx Chips
Xilinx, the world's leading provider of programmable solutions, today announced that its low-cost Spartan(TM) field programmable gate arrays (FPGAs) are on board the earth-friendly, biodiesel-powered Earthrace powerboat. The extraordinary vessel is the brainchild of New Zealander Pete Bethune, 41, a former oil exploration engineer. Bethune designed the 78-foot wave piercing powerboat in an attempt to break the world record for circumnavigating the globe (current record is 75 days) – and for the first time in history to do so using a powerboat that runs entirely on bio-diesel renewable fuel. Xilinx FPGAs enable the radar and night vision systems aboard Earthrace.
- Toshiba Tapes Out 90nm Consumer Wireless Chip with Synopsys IC Compiler
Synopsys, a world leader in semiconductor design software, today announced that Toshiba, a leading supplier of semiconductors, has used the Synopsys IC Compiler next-generation physical implementation solution to successfully tape out its next-generation TC90515XBG home digital network chip. Compared to its predecessor, the TC90515XBG delivers higher performance and increased functionality with reduced die-size and lower power consumption. The ability to use IC Compiler to concurrently optimize the design across all five of its operating modes was a critical factor in Toshiba's ability to meet its aggressive performance goals.
- Chassis Plans Speeds to Inc. 500 List
Inc. Magazine announced August 23 its 25th annual Inc. 500 ranking of the fastest growing private companies in America. Chassis plans, The Original Industrial Computer Source(TM), ranked number 151 on the list, with three year growth of 697%. Chassis Plans ranked first in the category of Computers and Electronic companies.
- Tundra Semiconductor Reports Latest Quarterly Revenue
Tundra Semiconductor Corporation (TSX:TUN), the leader in System Interconnect, today reported financial results for the first quarter of fiscal 2007, which ended July 30, 2006.
- National Debuts Precision Temperature Sensors with TruTherm Technology
National Semiconductor today introduced a family of high-precision, remote-diode temperature sensors that employ National's TruTherm(TM) thermal management technology with transistor-mode beta compensation. These new sensors are specifically designed for applications such as notebook and desktop computers and servers that are powered by high- performance microprocessors manufactured on 65 and 90 nm processes.
- Lattice Semiconductor Expands in India
Lattice Semiconductor today announced that it has established a sales force and forged partnerships with two major distributors to serve the programmable logic market in India. At a press conference in Bangalore today, Steve Donovan, Lattice corporate vice president of sales, introduced Vincent Su, Lattice Area Sales Manager for India, Korea, Taiwan and Southeast Asia; Rakesh Agarwal, who will be Lattice's Sales Manager, India; and Lattice's distribution partners, FE-Global Electronics (India) Pvt Ltd, Dragon Technology Ltd, and Spectra Innovations.
- Automotive Telematics to Grow to $38.3 Billion in 2011
Automotive telematics and navigation–including both hardware and services–will experience strong growth in several world regions, generating in total revenues of $38.3 billion in 2011, according to a new study released by ABI Research.
- Wireless Industry Partnership Launches WIP Portal
The Wireless Industry Partnership (WIP) today announced the launch of the WIP portal, a set of tools and resources for wireless companies around the world, available online. WIP estimates there are more than 40,000 wireless companies worldwide and designed these tools and resources in an effort to fuel business opportunities for member companies as well as to stimulate growth and innovation in the wireless industry.
- Agilent Introduces Parametric Test Systems with Linux Support
Agilent Technologies today announced that its 4070 Series Advanced Parametric Test Systems are the industry's first production parametric testers to support Red Hat Enterprise Linux. Because Linux is well established and runs on a PC platform, Agilent's customers can lower ownership costs and improve price performance. Agilent will continue to provide support for HP-UX 11i and offer easy migration to Linux for existing customers running HP-UX 11i, including SPECS and SPECS-FA.
- Xilinx Rolls Out PlanAhead 8.2 Design Tools for Virtex-5 FPGAs
Xilinx today announced immediate availability of the 8.2 version of its PlanAhead(TM) hierarchical design and analysis software with support for its newest Virtex(TM)-5 LX family of 65nm FPGAs. Used in conjunction with the Xilinx Integrated Software Environment (ISE(TM)) design tools, PlanAhead 8.2 software delivers a two-speed grade performance and cost advantage over competing offerings.
- Actel IGLOO FPGAs Lower Power Consumption
Actel today introduced the industry's lowest power field-programmable gate arrays (FPGAs), the Actel IGLOO(TM) family. With 5 µW static power, the flash-based family consumes 4x less static power than its nearest competitor and delivers more than 5x the battery life of the current leading PLDs in portable applications, thus setting a new bar for low power consumption. Because of the short product life cycles and competition in this marketplace, designers require a growing list of features and complexity, but not at the expense of draining the battery. This makes the reprogrammable, fully featured Actel IGLOO solutions attractive as an alternative to ASICs and CPLDs for portable applications.
- Sequence Creates Speedview-AMS to Extend EM And V-drop Analysis
Combining its award-winning Columbus-AMS extraction engine with electromigration and voltage-drop components from its CoolTime dynamic voltage-drop product, Sequence Design has created Speedview-AMS, the latest in a series of new tools for power-rail analysis. Speedview-AMS enables designers to diagnose EM and V-drop issues in full-custom designs.