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Recent studies have shown that the p70s6k/p85s6k signaling pathway plays a critical role in cell growth by modulating the translation of a family of mRNAs termed 5′TOPs, which encode components of the protein synthetic apparatus. Here we demonstrate that homozygous disruption of the p70s6k/p85s6k gene does not affect viability or fertility of mice, but that it has a significant effect on animal growth, especially during embryogenesis. Surprisingly, S6 phosphorylation in liver or in fibroblasts from p70s6k/p85s6k‐deficient mice proceeds normally in response to mitogen stimulation. Furthermore, serum‐induced S6 phosphorylation and translational up‐regulation of 5′TOP mRNAs were equally sensitive to the inhibitory effects of rapamycin in mouse embryo fibroblasts derived from p70s6k/p85s6k‐deficient and wild‐type mice. A search of public databases identified a novel p70s6k/p85s6k homolog which contains the same regulatory motifs and phosphorylation sites known to control kinase activity. This newly identified gene product, termed S6K2, is ubiquitously expressed and displays both mitogen‐dependent and rapamycin‐sensitive S6 kinase activity. More striking, in p70s6k/p85s6k‐deficient mice, the S6K2 gene is up‐regulated in all tissues examined, especially in thymus, a main target of rapamycin action. The finding of a new S6 kinase gene, which can partly compensate for p70s6k/p85s6k function, underscores the importance of S6K function in cell growth.
Cell cycle progression in response to mitogens requires the ordered activation of the cyclin‐dependent kinase (cdk) family of protein kinases and their cyclin partners (Sherr, 1996). However, for the cell to proliferate, it must also up‐regulate the biochemical machinery required to direct cell growth (Nasmyth, 1996). A chief component of the cell growth response is the generation of new translational machinery (Thomas and Hall, 1997), which is essential for the increased demand of the cell to perform an immense array of distinct anabolic processes (Nasmyth, 1996). Following mitogen‐induced exit from G0, the increased expression of many of the components of the protein synthetic apparatus has been demonstrated to be regulated at the translational level (see Meyuhas et al., 1996). Many of these transcripts are characterized by an oligopyrimidine tract at their transcriptional start site, or 5′TOP (Jefferies et al., 1994a; Terada et al., 1994; Jefferies and Thomas, 1996). Furthermore, recent studies have shown that the translational up‐regulation of these transcripts is mediated in part by activation of the p70s6k/p85s6k (Jefferies et al., 1997), presumably through 40S ribosomal protein S6, whose increased phosphorylation is hypothesized to facilitate the recognition of these transcripts by the 43S pre‐initiation complex (Jefferies et al., 1994a). The p85s6k isoform is expressed from the same transcript as p70s6k, through an alternative translational initiation start site (unpublished data), which adds a 23 amino acid nuclear localization signal (NLS) to the N‐terminus (Reinhard et al., 1994). The importance of these two kinases in cell growth has been inferred from either the use of the immunosupressant rapamycin (Chung et al., 1992; Kuo et al., 1992; Price et al., 1992) or through microinjection of neutralizing antibodies into cells (Lane et al., 1993; Reinhard et al., 1994), both of which selectively suppress mitogen‐induced p70s6k/p85s6k activation and impede cell growth.
The signal transduction pathway which mediates p70s6k/p85s6k activation has attracted a considerable degree of attention, because of its implied importance in cell growth and its potential use in identifying novel targets for immunosuppressive therapy (see Downward, 1998; Peterson and Schreiber, 1998). This pathway bifurcates at a growth factor receptor docking site that is distinct from that of the Ras–MAP kinase pathway (Ming et al., 1994), with activated phosphatidylinositide‐3 kinase (PI3‐K) as the proximal signaling component initiating downstream signaling (Chung et al., 1994). However, not all the data are consistent with this model nor with the role of other potential signaling components implicated in regulating p70s6k/p85s6k activation (see Downward, 1994, 1995; Thomas and Hall, 1997). A large part of this uncertainty stems from the complex nature of the p70s6k/p85s6k activation event, which relies on the sequential interplay between multiple phosphorylation sites (Ferrari et al., 1992; Pearson et al., 1995; Moser et al., 1997) and distinct intramolecular regulatory domains within the kinase (Pullen and Thomas, 1997; Dennis et al., 1998). Rapamycin blocks p70s6k/p85s6k activation through inhibition of mTOR/FRAP, a large molecular weight protein which is thought to serve as either a lipid or protein kinase (Thomas and Hall, 1997; Peterson and Schreiber, 1998). As treatment of cells with rapamycin selectively blocks p70s6k/p85s6k and S6 phosphorylation without affecting other protein kinases, it has been argued that p70s6k/p85s6k is the sole in vivo kinase responsible for regulating S6 phosphorylation. This conclusion was substantiated by the use of rapamycin‐resistant p70s6k/p85s6k mutants which protected S6 dephosphorylation from the bacterial macrolide (von Manteuffel et al., 1997). The potential impact of rapamycin treatment in immunosuppressive therapy combined with the importance of delineating mTOR/FRAP function has stimulated a great deal of interest in p70s6k/p85s6k (Thomas and Hall, 1997).
Although studies with rapamycin as well as inhibitory antibodies have led to the hypothesis that p70s6k/p85s6k plays an essential role in cell growth (Chou and Blenis, 1995), corroborative physiological studies have been lacking. Recently, however, we have identified a P‐element‐induced mutation in the Drosophila melanogaster homolog of the dp70s6k gene which severely impairs kinase expression and cell growth (unpublished data). Nevertheless, there is greater genetic diversity in mammals than in lower metazoans, and in many cases functional redundancy exists which is not revealed in less complex organisms (Miklos and Rubin, 1996). For these reasons, we set out to disrupt the p70s6k/p85s6k gene from mouse to evaluate its functional uniqueness in higher organisms and to determine which of the effects of rapamycin on cell growth are elicited through inhibition of p70s6k/p85s6k activity and S6 phosphorylation.
Disruption of the S6 kinase gene
Screening a mouse genomic library with a probe corresponding to the catalytic domain of a rat p70s6k/p85s6k cDNA (Kozma et al., 1990) resulted in the isolation of a 14 kb fragment of the mouse p70s6k gene (Materials and methods). To create a gene disruption by homologous recombination (Capecchi, 1989), we generated a targeting construct by replacing 1.2 kb of the genomic sequence encompassing the conserved Ser/Thr kinase catalytic subdomains VIII–X, with a neomycin resistance cassette (Figure 1A). Of 316 embryonic stem (ES) cell clones, three harbored the expected targeted mutation, as identified by PCR and Southern blot analysis (data not shown). One positive clone was used to establish chimeric mice that transmitted the targeted mutation through the germ line (Wood et al., 1993). Hetero‐ and homozygous mutant mice (Figure 1B) were obtained in Mendelian proportions, suggesting no obvious effect on viability. To confirm that the gene had been disrupted, Western blot analysis was performed with two distinct p70s6k antibodies which recognize either the N‐ or C‐terminus of p70s6k (Figure 1C). The results revealed a 50% decrease of the signal in heterozygous mice and no apparent signal in homozygous mice (Figure 1C). Similar results were obtained for the p85s6k isoform (data not shown). To test for S6 kinase activity, wild‐type, heterozygous and homozygous animals were injected intraperitoneally with cycloheximide, a treatment known to induce p70s6k activation in liver (Kozma et al., 1989). Liver extracts from these mice were assayed for p70s6k kinase activity by immunoprecipitation with either of the two antibodies described above. The S6 kinase phosphorylation assay revealed reduced activity in liver extracts from heterozygous as compared with wild‐type mice and no activity from homozygous mutant mice (Figure 1D). These results showed that the p70s6k/p85s6k gene had been disrupted and that the activity corresponding to this protein was absent from the homozygous mutant mice.
Phenotype of p70s6k gene disruption in mice
Three‐week‐old homozygous mutant mice were significantly reduced in body size when compared with wild‐type mice (Figure 2A). To ensure there were no variations due to in utero competition, body weights obtained at birth from either homozygous mutant or wild‐type crosses were compared (Table I). In both cases, the average body weight difference was ∼20% for both females and males. A comparison of homozygous mutant mice at 3.5 weeks of age demonstrated that the weights of all organs were proportional to the reduction in body weight (Figure 2B). Heterozygous mice were slightly smaller, although the values obtained were not significantly different from those obtained for wild‐type mice (data not shown). Analysis of body weight over an 11 week period following birth (Figure 2C) showed that up to ∼5 weeks of age both homozygous males and females grew more slowly than their wild‐type littermates. However, between 5 and 6 weeks of age, the growth rate for both homozygous and wild‐type offspring dramatically slowed down (Figure 2C), after which it appeared to be equivalent up to 11 weeks. The difference in body weight for both males and females was ∼15% at this stage. Thus disruption of the p70s6k/p85s6k gene resulted in a reduction in body weight and smaller animals.
Growth of embryos and primary embryo fibroblasts
As homozygous animals appeared to be retarded in their growth, embryos were analyzed at different stages in utero. The results revealed no obvious morphological differences between homozygous mutants and wild‐type animals at any stage of development. However, as depicted in a typical litter at 14.5 days of gestation, homozygous mutants were distinctly smaller in size, suggesting a growth delay without a corresponding delay in the rate of development. The difference in weight can be >30%, with heterozygotes displaying intermediate values (Figure 3A). To determine whether the effects on size were due to an impeded rate of cellular proliferation, 13.5 day mouse embryo fibroblasts (MEFs) from homozygous mutants and wild‐type animals were tested for their ability to proliferate in culture. Under the standard culture conditions employed, there was no difference in the ability of the two cell types to proliferate (Figure 3B). Furthermore, growth inhibition by rapamycin was similar in MEFs from both homozygous mutant and wild‐type mice (Figure 3C), suggesting that there was an additional target by which rapamycin exerted its inhibitory effects on cell growth in fibroblasts. Indeed, analysis of polysome profiles showed that loss of p70s6k/p85s6k does not affect the ability of mitogens to up‐regulate the expression of eEF1‐α, a typical 5′TOP mRNA (Figure 3D). Furthermore, rapamycin suppressed the up‐regulation of eEF1‐α to the same extent in MEFs derived from wild‐type and homozygous mutant animals (Figure 3D). Thus, the difference in embryo size was not reflected in the rate of MEF proliferation neither the effects of rapamycin on cell proliferation nor the translation of 5′TOP mRNAs could be attributed solely to p70s6k/p85s6k.
S6 phosphorylation in p70s6k‐deficient mice
Despite earlier studies indicating that p70s6k/p85s6k plays a critical role in the translation of 5′TOP mRNAs (Jefferies et al., 1997), there was no difference in this response in p70s6k/p85s6k‐deficient mice (Figure 3D). The effects of p70s6k on this response as well as on cell growth have been hypothesized to be mediated through increased S6 phosphorylation, and studies with rapamycin or dominant‐negative mutants of p70s6k have led to the conclusion that it was the sole kinase responsible for mediating this event in vivo (Chung et al., 1992; Jefferies et al., 1994a; von Manteuffel et al., 1997). To ensure that this response was abrogated in p70s6k/p85s6k‐deficient mice, the extent of S6 phosphorylation was examined in the livers of wild‐type and homozygous mutant mice following re‐feeding of starved animals, a protocol which previously had been shown to induce p70s6k activation and S6 phosphorylation in rats (Kozma et al., 1989). Two‐dimensional polyacrylamide gel analysis revealed that in starved wild‐type or mutant mice, S6 resided in the unphosphorylated form (Figure 4A), with only small amounts of the singly and doubly phosphorylated derivative, S6a and b, present. Re‐feeding of wild‐type mice induced a large increase in S6 phosphorylation (Figure 4A), with the appearance of all five of the phosphorylated S6 derivatives, S6a–e, clearly detectable. Surprisingly, in the liver of p70s6k/p85s6k‐deficient animals, the extent of S6 phosphorylation was comparable (Figure 4A). Similar results were also obtained if S6 phosphorylation was induced by intraperitoneal injection of cycloheximide (data not shown), which is thought to trigger increased S6 phosphorylation through inhibition of protein synthesis (Blenis et al., 1991). Consistent with these observations, serum stimulation of MEFs from either wild‐type or p70s6k/p85s6k‐deficient mice also induced full phosphorylation of the ribosomal protein, with most of S6 migrating in the position of the most highly phosphorylated derivatives S6d and e (Figure 4B). Furthermore, rapamycin pre‐treatment abrogated this response in MEFs from both wild‐type and p70s6k/p85s6k‐deficient mice (Figure 4B). The results demonstrated that in response to either re‐feeding or mitogen stimulation, S6 phosphorylation in the tissues examined was unimpaired in p70s6k/p85s6k‐deficient animals and remained sensitive to rapamycin.
A novel rapamycin‐sensitive S6 kinase activity
The results above implied that the mouse genome contained an unidentified rapamycin‐sensitive kinase capable of inducing S6 phosphorylation. To test this possibility, extracts from serum‐stimulated MEFs derived from wild‐type and p70s6k/p85s6k‐deficient mice pre‐treated in the absence or presence of rapamycin were analyzed by Mono S chromatography. Extracts from wild‐type mice showed a major peak of S6 kinase activity which eluted at ∼0.25 M NaCl and a less pronounced peak at 0.45 M (Figure 5A). Pre‐treatment of cells with rapamycin abolished both peaks of activity (Figure 5A). Previous studies had shown that the first peak represented the p70s6k (Jenö et al., 1988) and, as expected, the first peak of S6 kinase activity was absent in extracts from MEFs of p70s6k/p85s6k‐deficient mice (Figure 5A). However, the second peak of activity did not disappear and seemed to be slightly elevated in extracts from p70s6k/p85s6k‐deficient mice relative to wild‐type mice (Figure 5A), suggesting that the gene product may be up‐regulated in mice lacking the p70s6k/p85s6k gene. These results strongly indicated the presence of a novel and as yet unidentified rapamycin‐sensitive S6 kinase. Based on these observations, a search of the existing expressed sequence tag (EST) databases was initiated (Bugowski et al., 1993), resulting in the identification of a putative homolog of p70s6k/p85s6k (Figure 5B). We termed this homolog S6 kinase 2, (S6K2), and for clarity have termed p70s6k, S6K1 (Figure 5C). A comparison of the S6K1 sequence with that of S6K2 reveals that they were highly homologous, sharing 82% identity in the catalytic domain (Figure 5C). In addition, all the important regulatory domains are conserved, including the acidic N‐terminus, the linker region and the autoinhibitory domain, as well as the critical phosphorylation sites, including those equivalent to T229, S371, T389 and the S/TP sites in the autoinhibitory domain of S6K1 (Pearson et al., 1995; Moser et al., 1997). Clearly absent was the 23 amino acid NLS found in the p85s6k isoform (Reinhard et al., 1994), although a potential NLS resides at the C‐terminus of S6K2 (Figure 5B). Thus, the existence of a novel S6K was consistent with the regulated S6 phosphorylation identified in S6K1‐deficient mice.
Identification of S6K2
To determine whether the second peak of S6 kinase activity observed in extracts from wild‐type MEFs (Figure 5A) eluted in the same position as S6K2, extracts were prepared from serum‐stimulated 293 control cells or 293 cells that had been transiently transfected with a myc epitope‐tagged S6K2. The extracts were then analyzed by Mono S column chromatography as described above. The data showed that in comparison with extracts from control cells, the later eluting peak of S6 kinase activity was greatly enhanced in extracts containing the epitope‐tagged kinase (Figure 6A), in agreement with this peak representing S6K2. In addition, insulin‐induced activation of the transiently expressed myc‐S6K2 in 293 cells and this response was blocked by rapamycin (Figure 6B), consistent with the data presented above for MEF extracts from p70s6k‐deficient mice (Figures 4B and 5A). To ensure that the second peak represented S6K2, the equivalent Mono S peak fractions from liver extracts of p70s6k/p85s6k‐deficient mice, which had been subjected to cycloheximide treatment, were concentrated on an S6‐peptide affinity column (Materials and methods). This material was then analyzed for kinase activity and for the presence of S6K2 employing an antiserum directed against an N‐terminal sequence of S6K2 (Materials and methods). This antiserum, as compared with pre‐immune serum, specifically immunoprecipitated S6 kinase activity and detected a single band on Western blots which migrated at the predicted molecular weight of S6K2 (Figure 6C). Immunodepletion studies showed further that most of this activity could be removed from the pooled fraction after three successive immunoprecipitations, suggesting the existence of a single kinase (data not shown). Finally, RNase protection studies revealed that S6K2 transcripts appear to be ubiquitously expressed and increased in all tissues examined in S6K1‐deficient versus wild‐type mice (Figure 6D). The up‐regulation of S6K2 is especially high in thymus, where it reaches 3.8‐fold the level observed in wild‐type animals when normalized to a control mRNA (Figure 6D). These results suggested that in mice lacking S6K1, a compensatory mechanism leads to higher expression of S6K2, especially in the thymus where rapamycin had been shown to have a more pronounced effect on cell growth (Luo et al., 1994).
Here we demonstrate that S6K1‐deficient mice are viable and fertile, but they are significantly smaller in size (Figure 2), a phenotype particularly obvious in embryos (Figure 3). Surprisingly, the S6 phosphorylation response was unimpaired in livers of S6K1‐deficient mice following either a starvation/re‐feeding regime or injection with cycloheximide (Figure 4A). Furthermore, MEFs derived from S6K1‐deficient mice, as compared with wild‐type mice, displayed an equivalent sensitivity to rapamycin in terms of serum‐induced proliferation, 5′TOP mRNA translation and S6 phosphorylation (Figures 3 and 4B). These findings led to the identification of a new S6 kinase that functionally overlaps with S6K1 and whose transcripts are up‐regulated in S6K1‐deficient mice (Figure 6D).
Despite the presence of S6K2, S6K1‐deficient mice are smaller in size. This result provides further evidence for an involvement of the PI3‐K/S6K signal transduction pathway in growth control. It was already shown that disruption of upstream signaling components, such as the IGF receptor 1 (J.‐P.Liu et al., 1993) and the insulin receptor substrate 1 and 2 (IRS1 and 2) (Araki et al., 1994; Tamemoto et al., 1994; Withers et al., 1998), affect animal size. Moreover, in Drosophila, ectopic expression of PI3‐K recently has been demonstrated to promote growth of tissues by positively regulating both cell size and cell number (Leevers et al., 1996). It is clear that several signaling pathways, in addition to that of S6K, originate from the growth factor receptor–PI3‐K complex and are likely to cooperate in the control of growth. In the case of S6K1, the small size of homozygous mutant mice is consistent with a defect in translational capacity. Mutants have been described in Drosophila in which either expression of ribosomal proteins or rRNA are affected, termed Minutes or bobbed (Shermoen and Kiefer, 1975; Procunier and Dunn, 1978; Lindsley and Zimm, 1992). Either set of mutations leads to smaller flies which are delayed in development, similar to that reported here for S6K1‐deficient mice. Consistent with the role of S6K1 in translation, a P‐element insertion in the 5′ untranslated region (5′UTR) of the Drosophila homolog of the S6K gene, which severely suppresses the expression of the kinase, induces a Minute‐like phenotype (unpublished data). As S6K1 has been shown to be involved in regulating the expression of 5′TOP mRNAs at the translational level (Jefferies et al., 1997), a deficiency in the gene product would be expected to lead to reduced translational capacity and, consequently, the generation of smaller animals. Thus, it may be that in certain cell types or during specific development stages, the newly described S6K2 cannot fully compensate for the loss of S6K1 function with respect to S6 phosphorylation and the up‐regulation of 5′TOP mRNAs. To address this issue, a thorough analysis of these responses in specific organs and tissues throughout development will be required. In addition, the relative contributions of both genes in individual tissues would be greatly facilitated by generating mice containing a deletion of the S6K2 gene, as well as mice containing deletions of both genes.
Previous biochemical studies (Jenö et al., 1988) or extensive screening of cDNA libraries gave no indication of a second S6 kinase (Banerjee et al., 1990; Kozma et al., 1990; Grove et al., 1991). In addition, despite the high level of identity between S6K1 and S6K2, antibodies generated against S6K1 do not cross‐react with S6K2 (Figure 1). In most cases, S6K1 activity has been analyzed by Mono Q chromatography (Jenö et al., 1988). Under these conditions, we have found that both kinases eluted in the same fraction (data not shown). Furthermore, in the initial purification of S6K1, Mono S chromatography was not employed until the final step (Jenö et al., 1988, 1989); at this point S6K2 may have been separated from the S6K1 active material. This fact, along with preliminary data showing that S6K2 is more sensitive to phosphatases than S6K1 during extraction (M.Sanders and M.Pende, unpublished), may explain why S6K2 was not detected in earlier biochemical studies. Here we show that Mono S chromatography of MEFs and 293 cell extracts reveals both peaks of S6K activity. The major peak of activity corresponds to S6K1, and the second peak represents S6K2. These data, together with extensive searches of EST databases, suggest that there are no additional homologs of functional S6 kinases in mice. It also appears from the Mono S chromatography profile that S6K1 is the more dominant activity; however, the in vitro assay conditions employed may not be optimal for detection of S6K2 activity. Furthermore, as stated above, S6K2 appears to be more sensitive to phosphatases. Higher in vivo S6K2 activity might explain the robust S6 phosphorylation response observed in the livers or fibroblasts derived from S6K1‐deficient mice. It is also clear that S6K2 transcripts are up‐regulated in S6K1‐deficient mice (Figure 6D) and the same may be true for protein levels; however, to resolve this latter point will require higher affinity antibodies. Another possibility is that protein phosphatase 1, which selectively dephosphorylates S6 in vitro (Olivier et al., 1988), could also be suppressed in S6K1‐deficient mice, compensating for the loss of S6K1 and leading to higher levels of S6 phosphorylation. Evidence for such a mechanism has been demonstrated in cells transformed by temperature‐sensitive Src, where high levels of S6 phosphorylation are largely maintained through suppression of protein phosphatase 1 activity rather than S6K activation (Belandia et al., 1994). Future studies employing the use of high affinity S6K2 antibodies as well as assays designed to measure the impact of protein phosphatase 1 on the S6 phosphorylation response will be important in resolving whether such compensatory mechanisms are operating in S6K1‐deficient mice.
The identity between S6K1 and S6K2 is >80% in the catalytic domain, suggesting that these forms result from gene duplication of the type described for Hox and tyrosine kinase genes in vertebrates (Spring, 1997; Aparicio, 1998). The high degree of S6K1/S6K2 identity in the catalytic domain extends to the linker region where critical phosphorylation sites involved in regulating S6 kinase activity reside, including S370 and T388. Indeed, of the eight phosphorylation sites identified in endogenous S6K1, all, as well as their surrounding motifs, are present in S6K2 (Figure 5). In addition to these sites, phosphorylation of S17 (Weng et al., 1995) as well as of T367 and T447 (Pearson et al., 1995) has been identified in overexpressed S6K1. Of these sites, only T447 is absent in S6K2. The homologous site to T388, T389 in S6K1, is the principle target of rapamycin‐induced dephosphorylation and inactivation of S6K1 (Pearson et al., 1995). This effect requires the acidic N‐terminus of S6K1 (Dennis et al., 1996), which is also present in S6K2, consistent with the fact that S6K2 is also sensitive to rapamycin (Figure 6B). Noticeably absent in S6K2 is evidence of a nuclear targeting sequence, such as that found in S6K1. However, as the 5′UTR of the S6K2 gene does not reveal in‐frame stop codons, a nuclear‐targeted S6K2 isoform could still be generated by an alternative translational start site at an upstream codon. In addition, there is a basic sequence at the C‐terminal end of S6K2 which could also serve as an NLS (Figure 5B). Additional studies employing better antibodies to the endogenous kinase will be required to resolve this issue. The other striking feature of S6K2 is a sequence extending from residue 435 to 459, which contains 50% prolines and is noticeably absent in S6K1. A search of potential related sequences showed that residues 453–469 are quite homologous to a domain in the p85 subunit of the PI3‐K which has been shown to bind to Src kinase (X.Liu et al., 1993). If this motif acts as a docking site for an SH3‐containing protein, it would suggest a role for S6K2 distinct from that of S6K1 in downstream signaling.
During the completion of these studies, it was reported that deletion by homologous recombination of the S6K1 gene in ES cells led to the generation of a clonal line which exhibited a 50% slower growth rate and maintained rapamycin sensitivity (Kawasome et al., 1998). In this case, no phosphorylation of S6 was detected in homozygous deleted S6K1 cells versus either wild‐type or heterozygous cells. Furthermore, 5′TOP transcripts were present on polysomes in the absence of mitogens, whereas these transcripts were largely present in mRNP particles in wild‐type cells, as previously reported (Jefferies et al., 1994a; Terada et al., 1994; Jefferies and Thomas, 1996). The slower growth rate of these embryonic stem cells could be viewed as being consistent with the apparently slower rate of growth exhibited by S6K1 embryos. Although the initial S6K2 EST was derived from a mouse blastocyst library (Lennon et al., 1995) and we have detected S6K2 transcripts in every other cell type we have examined, it may be that S6K2 is non‐functional in ES cells. However, it should also be noted that the method employed by Kawasome et al. (1998) to examine S6 phosphorylation was 32P‐labeling of growing cells. Under these conditions, S6 phosphorylation is normally quite low (Nielsen et al., 1981). Future studies should resolve these differences.
In most cell types, rapamycin delays cell proliferation whereas in T cells it abolishes this response (Kuo et al., 1992); this effect is also observed in vivo (Luo et al., 1994). This selectivity for T cells is being exploited in immunosuppressive therapies following organ transplantation (Borman, 1994). We have found that the level of S6K1 is particularly high in thymus. However, we have seen no difference in T cell proliferation induced by co‐stimulation with CD28/CD3 antibodies in cells derived from either S6K1‐deficient or wild‐type mice (unpublished data). Nevertheless, the extent to which S6K2 transcripts are up‐regulated in S6K1‐deficient mice is most dramatic in thymus, suggesting an important role for S6K in this organ. Disruption of the S6K2 gene in the S6K1‐deficient background and the use of RAG‐2 mice (Chen, 1993) should resolve the question of the importance of S6K in T cell proliferation and its value as a potential therapeutic target for transplantation therapy.
Materials and methods
Construction of the targeting vector
By screening a 129/SV mouse genomic library (Stratagene) with p70s6kcDNA as a probe, we obtained a 14 kb fragment that contained the sequence of the catalytic domain of p70s6k. The 3.4 kb NheI–ApaI fragment and the 1.0 kb SpeI–PstI fragment of the p70s6k/p85s6k gene were ligated into the XbaI and SalI sites of the neo cassette vector (Arber et al., 1997), respectively. The resulting targeting vector was designated pS6Kneo/TK.
Electroporation of embryonic stem cells
The pS6kneo/TK targeting vector (50 μg) was linearized at the single NotI site and electroporated into 1.0×107 E14 ES cells at 500 mF and 250 V. The cells were then cultured in the presence of G418 (0.6 mg/ml) and gancyclovir (2 μM) for 10 days. Homologous recombination events were identified by PCR. The forward primer for the PCR was 5′‐TGCTCTCAGTCATGGTCTCCACCAA, which corresponds to the sequence of the 3′ end of the neo cassette. The reverse primer was 5′‐AACGCACGGGTGTTGGGTCGTTTG, which corresponds to the sequence of the p70S6k gene flanking the short arm of the targeting vector. The primers were used according to the following cycling conditions: 3 min at 94°C (one cycle); 0.5 min at 94°C, 0.3 min at 67°C and 1 min at 72°C (40 cycles); 10 min at 72°C (one cycle). ES cell colonies were screened by PCR for the 1.1 kb fragment specific of the targeting event.
Chimera generation and Southern blot analysis
Aggregation of positive ES cells with morula‐stage embryos obtained from inbred (C57Bl/6×DBA/2) F1 mice was performed following Wood et al. (1993). Chimeric mice were crossed to C57Bl/6 mice and germline transmission was assessed by Southern blot analysis. DNA extracted from the tails of mice was digested with BamHI, and 10 μg was loaded onto a 0.7% agarose gel and transferred to a nylon membrane (Porablot, Macherey‐Nagel). Hybridization and washing were performed as previously described (Reinhard et al., 1992). The probe used for hybridization was the 300 bp PCR product amplified with the forward primer, 5′‐ACAGTGGATGAGCCTACTTG, and the reverse primer, 5′‐GGACGGAGGTATCAGTGTAG.
Isolation of mouse embryo fibroblasts, cell cultures and two‐dimensional gel electrophoresis
Day 13.5 embryos were minced and incubated in 0.25% trypsin at 37°C for 15 min, and then filtered through a cell strainer (Falcon) as previously described (Blasco et al., 1997). Dulbecco's modified Eagle's medium (DMEM), containing fetal calf serum (FCS) at a final concentration of 10%, was then added to the cell suspension. Cells were then centrifuged at 1000 r.p.m. and the pellet was suspended in DMEM containing 10% FCS. To measure cell doubling times and saturation densities, cells were plated in 6‐well Falcon plates at a density of 2.5×105 cells per well. Cells were maintained and counted every day after trypsinization. For measurements of [3H]thymidine incorporation, cells were plated in 24‐well Falcon plates at a density of 0.5×105 cells per well and cultured in the presence of [3H]thymidine (1 mCi/ml) and increasing concentrations of rapamycin for the indicated times. The [3H]thymidine incorporation was measured by liquid scintillation counting. Ribosomal proteins were isolated from either mouse liver or MEFs, and the level of in vivo S6 phosphorylation was examined by two‐dimensional polyacrylamide electrophoresis as previously described (Olivier et al., 1988).
Polysome profiles and Northern blot analysis
Cytoplasmic extracts for polysome fractionation were prepared as already described (Jefferies et al., 1997). The extracts were run on a 17.1–51% sucrose gradient in an SW60 Beckman rotor at 56 000 r.p.m. for 58 min at 4°C. Each sample was then fractionated into 12 fractions. Isolation of RNA and Northern blot analysis of the fractions were performed as described previously (Jefferies et al., 1994b).
Cloning of S6K2 cDNA
A search in the EST database (Lennon et al., 1995) for p70s6k homologs provided us with the partial sequence of three clones (accession Nos AA563175, clone 975227; AA396470, clone 569218; and AA103535, clone 556081). The first clone showed similarity with the 5′ end of p70s6k, whereas the other two clones had overlapping sequence and were similar to the 3′ end of p70s6k. This sequence information was used to design two primers complementary to the predicted ends of a putative p70s6k homolog. The sequence of the forward primer was 5′‐CAGAAGCTTATCTCCGAGGAGGACCTGGCGGCCGTATTTGATTTAGACTTGG and that of the reverse primer 5′‐ATAAGAATGCGGCCGCTGCAGTTCTCACAGCTGCCCTCTCTTCCTCTATTCTCCTAACG. A 1.5 kb fragment was generated by PCR using these primers and cDNA from a mouse muscle library as a template (Stratagene). The 1.5 kb fragment was then amplified with the same reverse primer and the forward primer 5′‐AGCTCGCGAGGATCCGTGGTTATGGAGCAGAAGCTTATCTCCGAGG. The nested PCR introduced the sequence corresponding to the myc 9E10 epitope (Evan et al., 1985) after the initiation codon of S6K2. The myc‐S6K2 fragment was subloned into the BamHI and the NotI sites of a human cytomegalovirus promoter‐driven expression vector and sequenced. Clone 975227 was obtained from American Type Culture Collection (ATCC) and entirely sequenced. The sequence of clone 975227 and the myc‐S6K2 sequence were compared, and the sequence of the S6K2 open reading frame was registered in the DDBJ/EMBL/GenBank database under the accession No. AJ007938.
Antibody production
The peptide MAAVFDLDLETEEGSEGEGEPEFSPADV, comprising the first 28 amino acids of S6K2, was coupled to a C‐terminal KKC peptide, conjugated to keyhole limpet hemocyanin and used to produce rabbit polyclonal antiserum (Neosystem, Strasbourg, France).
Transient transfections, immunoprecipitation, immunoblotting and kinase assays
Human embryonic kidney 293 cells were maintained, transfected by a calcium phosphate precipitation method and stimulated as previously described (Dennis et al., 1996). Protein concentrations were measured using the BioRad D/C protein assay. For Western blot analysis, either 50 μg of protein extract from thymus or 20 μg of protein extract from cells was resolved by SDS–PAGE before transfer onto an Immobilon P membrane (Millipore). Endogenous p70s6k was monitored with either the M1 or M5 antibody as previously described (Lane et al., 1992). Expression of the epitope‐tagged S6K2 was detected with the monoclonal 9E10 antibody as previously described (Ming et al., 1994). Expression levels were quantified using fluorimetry (Molecular Dynamics) and ImageQuant software (Molecular Dynamics). Kinase activity assays and quantitation of results were carried out as previously described (Pullen et al., 1998).
Protein extracts were prepared and fractionated by Mono S cation‐exchange chromatography as previously described (Lane and Thomas, 1991). Briefly, cells were washed twice with ice‐cold extraction buffer [15 mM pyrophosphate, pH 6.8, 5 mM EDTA, 1 mM dithiothreitol (DTT), 1 mM benzamidine and 0.1 mM phenylmethylsulfonyl fluoride (PMSF)], scraped from the culture dish and homogenized with 12 strokes of a Dounce homogenizer. To remove cell debris, homogenates were first spun at 8000 g for 10 min at 4°C followed by ultracentrifugation at 340 000 g for 1 h at 4°C. Supernatants were filtered through a 0.22 μm filter and loaded onto the Mono S column, pre‐equilibrated with Mono S buffer (50 mM MOPS, pH 6.8, 1 mM EDTA, 10 mM pyrophosphate, 1 mM DTT, 1 mM benzamidine and 0.1% Triton X‐100). The column was washed with Mono S buffer and developed with a 15 ml linear salt gradient up to 0.5 M NaCl, at a flow rate of 0.5 ml/min. Fractions of 0.5 ml were collected and assayed for S6 kinase activity at a 1:10 dilution (Lane and Thomas, 1991). For immunoprecipitation and Western blot analysis, fractions containing S6 kinase activity were pooled, diluted with 15 vols of peptide buffer (20 mM diethanolamine, pH 8.5, 1 mM EDTA, 1 mM benzamidine, 1 mM DTT, 15 mM pyrophosphate, 0.1% Triton X‐100) and concentrated on a 50 μl affinity column having as a solid phase S6 peptide (20 residue peptide S6228–249) coupled to activated CH–Sepharose 4B (Ferrari, 1998). The column was then washed with 6 vols of peptide buffer containing 0.2 M NaCl, and proteins were eluted with 200 μl of elution buffer (20 mM Bis‐Tris, pH 6.3, 1 mM EDTA, 1 mM benzamidine, 1 mM DTT, 15 mM pyrophosphate, 0.5 M NaCl, 0.1% Triton X‐100). A 50 μl aliquot of the eluate was analyzed on Western blots using the anti‐S6K2 antibody described above. A second 10 μl aliquot was immunoprecipitated with the S6K2 antibody, and S6 kinase activity of the immune complex was assayed as described (Lane et al., 1992).
RNA extraction and RNase protection assay
Total RNA from mouse tissues and cultured cells was extracted as previously described (Chomczynski and Sacchi, 1987). To obtain an S6K2‐specific probe, the last 171 nucleotides of the S6K2 coding sequence were amplified by PCR using primers that introduced a BamHI and an EcoRI site, respectively, at the 5′ and 3′ ends. The PCR fragment was subcloned into the BamHI and the EcoRI sites of pBluescriptKS. The resulting plasmid was then linearized with BamHI and served as a template to generate, by in vitro transcription with T3 polymerase, a radiolabeled antisense S6K2 RNA probe of 243 nucleotides. The S6K2 probe (105 c.p.m.) was added to 10 μg of total RNA and dried in a speed‐vac. RNA was resuspended in 10 μl of hybridization solution (40 mM PIPES, pH 6.7, 400 mM NaCl, 1 mM EDTA and 80% formamide), incubated for 3 min at 95°C and for 16 h at 45°C. Samples were then incubated for 1 h at 28°C with 100 μl of RNase solution (10 mM Tris, pH 7.5, 200 mM NaCl, 5 mM EDTA, 100 mM LiCl, 30 μg/ml RNase A and 6 U/ml RNase T1). RNase digestion was terminated by adding 2 μl of 10% SDS and 2 μl of 20 mg/ml proteinase K. After incubation at 37°C for 15 min, RNA was extracted with phenol/chloroform and precipitated with ethanol using 20 μg of tRNA as carrier. The RNA pellet was washed with 70% ethanol, resuspended in formamide loading buffer and loaded on a 5% polyacrylamide gel containing 8 M urea. At the end of the run, the gel was dried and analyzed by PhosphorImager (Molecular Dynamics).
We thank T.Ochiya and T.Matsuyama for their guidance during the p70s6k/p85s6k disruption studies, P.Kopf and J.‐F.Spetz for their assistance with ES cells and embryonic aggregation, and S.Ferrari for providing S6 peptide affinity ligand. We are also grateful to P.Caroni, P.Dennis, U.Muller, N.Pullen and S.Volarevic for critical reading of the manuscript, and M.Rothnie for preparing figures. We thank Melanie Sticker and Melissa Sanders for their technical help. M.P. and S.F. are supported by stipends from the EEC and EMBO/HFSPO, respectively. These studies were supported in part by grants from the EEC and HFSPO to G.T.
- Copyright © 1998 European Molecular Biology Organization

subset: commoncrawl


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From Lagers to Ales, beer is a universal drink and is enjoyed by many all over the world. Since beer comes in different colors and flavors it's practical to learn about the varieties and discover which one you like best. Not only to stock up on your favorite beer but also to share it with your friends and family.
Let's start with the different varieties of beer. The beer you are drinking can be either Lager or Ale. Within these two groups are different varieties to choose from. Let's explore them, shall we?
The term "Lager" refers to storing the beer at cool temperatures. Lager is considered as the base of all beers and usually serves as a standard to measure other beers. After a brewing process of four to twelve weeks, it results in a smooth beer with crisp taste. The lager category consists of "Pale Lager" and "Dark Lager".
Pale Lager
Pale Lagers are the most consumed and commercially available style of beer in the world. With their golden color, they possess a clean very subtle "hoppy" flavor. The main ingredients are water, malt and noble hops. The noble hops are the source of the dry bitterness of the beer. Other ingredients occasionally added are corn or rice, to lighten the body of the beer.
Dark Lager
Dark lagers come in colors ranging from copper to dark brown. Their dark color comes from the extra caramel that is added to the drink or the type of malt that is used. It has a low level of hops strength and bitterness. The taste tends to lean more toward sweet hints of caramel, chocolate and nuts.
Pilsner has an almost transparent golden color and claims to be the oldest variety of Lager. It has by far the most intense bitter taste from the hops. However, the taste of pilsner differs between the production countries. For example, the Czech pilsner has a lighter flavor, while the German pilsner is more bitter and earthy.
The term "Ale" was originally used to describe a "hopless" brewed drink, however throughout the years, the meaning changed to "lightly hopped" drink. This is exactly what ale beers are. Hops are added to impart a bitter herbal flavor that balances the sweetness of the malt. Because of the high temperature during fermentation, the speed in which the yeast ferments the beer increases, giving it a sweet, full bodied and fruity taste.
Brown Ale
Brown Ale has a dark amber or brown color. This variety of Ale is mostly popular in England and North America. When produced in North America it tends to be dry with slight citrus hints. The sweet and chocolaty aroma, bitterness and medium body are drawn from the American hop type. In England the taste ranges from malty, nutty to sweet with low alcohol. The taste depends if it's from Northern or Southern England.
Pale Ale
Pale Ales are made from malt that is dried with coke, which is a type of coal. The malt used for this type of beer is called, pale malt. That is why it results in a lighter colored, pleasant beer. Pala Ale has a very high level of hops, making it nice and bitter.
Flavors of Beer
The flavors that are present in beer generally described in three terms. By learning to identify these flavors you will be able to decide what balance of these traits you prefer. Then you'll be well on your way to ordering beer like a true expert.
The term maltiness is used to describe the sweetness of the beer. Malt is derived from grains, and is the part of the beer that is fermented. The grains that are generally used to create malt are wheat and barley. There are several varieties of malt, with varying flavors.
Hops are the plants used during production of beer to add the bitter flavor. Bitter might sound unpleasant, but actually it is not hard to appreciate. Pale ales are generally the hoppiest beers available. Another way to recognize high level of hops in you beer is the aroma of fresh herbs, grapefruit and pine needles.
Actual fruit are not added to the production of beer, but their flavors can be tasted in the beer. How is that possible? Well during fermentation, not all sugars are consumed; these leftovers provide the beer with a sweet taste, usually very similar to the taste of fruits. The fruitiness in beer is reminiscent of pears, apples and bananas.


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Kim writes:
I think I spotted an error in the Harvard Gazette, of all places. Check out this headline. I always thought awhile was one word.
Thanks for sending this in, Kim! You're right. There is an error, but it's not what you're thinking. There is awhile, and there is also a while. Their meanings are so close that many people use them interchangeably, but there is a difference between them.
Where does the Harvard Gazette go wrong? It uses the noun, a while, instead of the adverb, awhile.
Quick rules:
- A while is a noun meaning "a short spell." It has been a while since I met with Tom.
- Awhile is an adverb meaning "for a short spell." Use it to modify verbs. Let's wait awhile. After school, I needed to sleep awhile.
- When there is a preposition, use a while. (A noun can be the object of a preposition, but an adverb can't be.) Let's sit down and talk for a while. We spoke a while ago.
Try this:
- When in doubt, try replacing awhile with another adverb, such as "slowly" or "happily." If the substitute adverb sounds fine, then use awhile. If not, use a while.
Let's wait quietly. (It works.) >> Let's wait awhile.
Let's wait for quietly. (It doesn't work.) >> Let's wait for a while.
- Sit down, stay awhile
- Sit down, stay for a while


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Is Fossil collecting for me ? Are you interested
in the way the world used to be ? Do the strange creatures and
plants that used to inhabit earth fascinate and interest you ? If
so, then fossil collecting is for you. Fossil collecting brings
you closer to these life forms, and handling the real thing can
teach you more than most books can.
How do I get started ? The best way you can start off your fossil collection is to join a club, and get contact with other fossil collectors. They will help you with how and where to find fossils in your local area and may provide useful contacts for the future. Some kind people will even give you a few of their swaps, just to get your collection started.
Where will I keep all these fossils ? There are a lot of ways you can keep your fossils. Depending on the size of them, you can store fossils in labeled boxes with cotton wool. Wooden slide out drawers with different size compartments are very handy, as are display cabinets. A good fossil collection should not be kept hidden but displayed for all to admire. Shelves displaying fossils should not be cluttered, but kept simple. Different material that can be used to good effect are glass, wood and shiny metals. Display stands can be expensive brand new, so have a look around local secondhand stores for a set of shelves that would be best to display your particular collection. Fossils should be dusted every month or so, and labels with their name, location and collector, should be kept with the specimen at all times. Unlabeled fossils are worthless to many collectors and scientists.
Where are fossils ? This really depends on where you live. Fossils can be found in a lot of different places, such as on the sea floor, in creeks / rivers, on beaches, in cliff sides, in quarries, or even beside roads. Fossils are only found in sedimentary rocks formed over 10 000 years ago. Fossils from the sea are plentiful as shells are easily covered by sediments. To find fossils in your area, you should once again talk to experienced collectors, or consult a geological map. You will find out a lot about the ancient history of your area and all the old coastlines etc.
How do I get fossils from the site to my collection ? You may be lucky enough to find fossils free of matrix (surrounding rock) but sooner or later you will have to remove a fossil from it. If it is a large boulder, a sledgehammer can be used, a smaller piece can be handled with hammer and chisel and small, delicate pieces with a fine scratching tool or acids. The specimen can then be washed, labeled and displayed. Protective coatings of plastics can be used to protect rare or good condition fossils.
The above is
only a short guide but other, more comprehensive books should be
consulted. One book I recommend for beginners is : The
Nature Company Guides : ROCKS AND FOSSILS by
Arthur B. Busbey III, Robert R. Coenraads, Paul Willis, David
Roots. RD Press , Australia, 1996
BARRY VAN BAKEL : email@example.com


text:
Modular SPM system
MultiView 4000
The MultiView 400 is an extremely versatile and modular SPM system. Indeed, this instrument is the only SPM system available with a free optical axis from both above and below the sample, and thus can be moved easily between different host microscopes allowing for completely integrated microscopy...
Scanned probe microscopy and conventional optical microscopy
MultiView 1000
The award winning MultiView 1000™ is the first system available that fully integrates all forms of scanned probe microscopy with conventional optical microscopy. Designed around Nanonics' patented, award winning 3D Flatscan™ scanner technology and incorporating sophisticated cantilevered optical fiber probes, the instrument can simply and transparently be combined with any inverted, upright, or dual optical microscope....
SNOM probes
Nanonics is the only supplier of the entire spectrum of probes used in SNOM today. Those probes perform all mode of NSOM operation and are breaking new ground with exciting new techniques...


text:
On campus, we have a specialist team based within our Careers & Employability Centre on the ground floor of the University Library, and this is an ideal place to start. We are open 9am until 4.30pm Monday to Friday where you can drop in with any careers queries you may have. If it's a quick enquiry such as a CV check, or advice on personal statements, job applications or job search, you can just pop in to our Careers Clinic between 1-4pm. If travelling to visit us, please give us a quick call to double check that we're open.
We offer a comprehensive events programme giving you the opportunity to hear from the experts. With a host of employers and external organisations offering bespoke sessions to support you, these are not to be missed. Furthermore, our annual Careers Fair, 'Find Your Feet' is a great opportunity to network with the employers of the future. Group advice sessions take place throughout each week and cover a variety of topics, from CV writing to job search techniques and interview preparation; these can all be found on the Events page.
Our vacancy service will give you access to 1000s of graduate level jobs, as well as work placements and part time jobs for whilst you are studying here, helping you to set yourself aside from the competition, in an increasingly competitive graduate labour market.
To help you make sense of all of these activities, we offer you the opportunity to work towards your Lincoln Award, to help you to understand how you can apply your learning and experiences to the 'world of work'.
In addition to all of this we offer one to one careers guidance, and subject specific workshops and lectures, so look out for us in your School's, but should you want any information on any of our services please do get in touch by visiting our Careers Centre.
Finding your career can be tough, but we are here to help guide you through the challenges and to help you to achieve your full potential and ambition.


text:
At SME Web Design we pride ourselves on developing technically superior websites. Our grasp of search engine optimisation and databases, combined with our web design and hosting expertise, has allowed us to succesfully undertake projects of quite varied complexity. In recent times this has enabled us to develop a unique range of website specifications, which offer exceptional value for money.
We have four website specifications, ranging from budget to ecommerce, all of which are search engine optimised, use modern CSS design, are W3C compliant, include Google Analytics, Google Sitemap, are customisable and upgradable. We also have a number of customisable specialist website systems and work with experts in sql server, high quality video, My Presenter videos, and 360 imagery.
Search Engine Optimised Website Specifications
- Budget
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- Database
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Budget Search Engine Optimised Website
Modern CSS Business Template webpage layout, with a Home page optimised for your business name, plus Contact Us page incorporating a unique anti-spam enquiry form. Google Analytics and Google Sitemap included. Top quality website hosting and support. Fantastic value - find out more ...
Advanced Search Engine Optimised Website
Expertly designed, W3C compliant, up to 10 pages, branded with your own logo and images. Webpages optimised for Google based on chosen keyword phrases. Google Analytics, Google Sitemap, and a unique anti-spam contact form all included. Top quality website hosting and much more ...
SQL Server CMS Search Engine Optimised Website
Includes all the features of the Advanced Specification, up to 20 pages, plus your own SQL Server Database and fully customised Content Management System. Update your web pages, create news article webpages, site/download links, and carry out your own on-page search engine optimisation, plus much more ...
Ecommerce Search Engine Optimised Website
Includes all the features of the Advanced and Database specifications, plus your own e-commerce store with shopping cart, checkout, and payment service provider functionality. Using our unique method individual product web pages are highly optimised, for the search engines, based on their product titles, plus much more ...
Specialist SQL Server Driven Websites & Applications
We have a number of database/website systems, which can be customised for clients on very competitive terms. Currently these include; training, job recruitment, holiday rentals, and cricket leagues. We also have a number of advanced website components that can be added to existing sites. Please call for further details.
Specialist Partner Services
At SME Web Design we are constantly on the lookout for opportunities to network with specialists that can provide high quality content and applications that can be incorporated in to for our clients websites. This currently includes:-


text:
Hawkins said, "When I ran for District Councilor in 2011, people wrote me in for every council seat. At large. I think I came in second in the District Attorney's race. They were writing me in for judge. A lot of people want to vote for the Greens and vote for me."
Updated 05/17/2013 05:14 PM
Uncertainty in the race for Syracuse mayor
The outlook remains unclear in this year's race for mayor in Syracuse. Just who will end up on the ballot in November? YNN's Bill Carey says as the political parties work to finalize their slate of candidates, there are still a lot of variables at play.
SYRACUSE, N.Y. -- Tom Dadey and Stephanie Miner don't agree on much, but they share growing concerns over this year's race for mayor. Miner is seeking re-election and hoping to avoid any battles within her own party.
"I think primaries are divisive. I think our city has significant challenges and that we should focus on those challenges and not focus on primaries and wasting resources," Miner said on Wednesday.
That may be wishful thinking. Already, one democrat, Alfonso Davis, says he will challenge the incumbent mayor in the September primary. And there is ongoing speculation about Common Councilor Pat Hogan, a democrat, who says he is still mulling over a primary challenge.
Republican chairman Dadey thinks the struggles inside the democratic party offer an opportunity to retake the mayor's office. His problem is finding someone to run. It has not been easy.
Dadey said, "It is very costly to raise a lot of money to run in a very competitive race against a very well-funded incumbent. The time, the sacrifice, the commitment it takes."
The only republican actively considering a candidacy is former Syracuse firefighter Tom Sexton. But Dadey says he is not the only possibility.
"My goal would be to have a republican candidate for mayor. Now, if that's not possible, we may look at an alternative. Whether it's a democrat who can agree with some of our party principles or somebody else who's possibly a non-enrolled voter or enrolled in another party. We may not rule out that option as well," Dadey said.
Dadey, for months, has been considering whether to give democratic Councilor Hogan the republican line to bolster his challenge to Miner.
And in the midst of all that, there is a new wild card. The Green Party has a guaranteed ballot line. And one of the party's most prominent names is thinking about a potential run for mayor. Howie Hawkins, who ran as the Green Party candidate for governor in 2010, says a decision will come soon.
Hawkins said, "When I ran for District Councilor in 2011, people wrote me in for every council seat. At large. I think I came in second in the District Attorney's race. They were writing me in for judge. A lot of people want to vote for the Greens and vote for me."
Hawkins expects to announce a decision, on a run for mayor, next week.
Republican committee members meet this weekend to choose candidates for City of Syracuse races, but may delay a decision on the mayoral contest. The democrats have already designated Stephanie Miner. The Green Party meets for its selection process on Monday.
People challenging those results can begin circulating petitions on June 4th to force a primary in September.


text:
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©PornMD.com, 2019


text:
"It's telling that, amid all the recent recriminations, even lenders have not fingered CRA. That's because CRA didn't bring about the reckless lending at the heart of the crisis. Just as sub-prime lending was exploding, CRA was losing force and relevance. And the worst offenders, the independent mortgage companies, were never subject to CRA — or any federal regulator. Law didn't make them lend. The profit motive did."-Robert Gordon, American Prospect
I have been meaning to get back to this issue, but events in the market have kept me a tad busy.
Making the rounds amongst a certain subset of wingnuts on CNBC, at IBD and other selfconfoozled folks has been the meme that the entire housing and credit crisis traces to the the Community Reinvestment Act (CRA) of 1977. An alternative zombie myth is the credit crisis is due to Fannie Mae and Freddie Mac. A 1999 article from the New York Times about the GSE's role in subprime mortgages has been circulating as if its the rosetta stone of the credit crisis.
These memes have become a rallying cry — cognitive dissonance writ large — of those folks who have been pushing for greater and greater deregulation, and are now attempting to disown the results of their handiwork.
I feel compelled to set the record straight about this pseudo-intellectual detritus. As we have painstakingly discussed over the past few years, there were many direct and indirect causes of the current financial mess.
Let's clarify the causes of current circumstances. Ask yourself the following questions about the impact of the Community Reinvestment Act and/or the role of Fannie & Freddie:
• Did the 1977 legislation, or any other legislation since, require banks to not verify income or payment history of mortgage applicants?
• 50% of subprime loans were made by mortgage service companies not subject comprehensive federal supervision; another 30% were made by banks or thrifts which are not subject to routine supervision or examinations. How was this caused by either CRA or GSEs ?
• What about "No Money Down" Mortgages (0% down payments)? Were they required by the CRA? Fannie? Freddie?
• Explain the shift in Loan to value from 80% to 120%: What was it in the Act that changed this traditional lending requirement?
• Did any Federal legislation require real estate agents and mortgage writers to use the same corrupt appraisers again and again? How did they manage to always come in at exactly the purchase price, no matter what?• Did the CRA require banks to develop automated underwriting (AU) systems that emphasized speed rather than accuracy in order to process the greatest number of mortgage apps as quickly as possible?
• How exactly did legislation force Moody's, S&Ps and Fitch to rate junk paper as Triple AAA?
• What about piggy back loans? Were banks required by Congress to lend the first mortgage and do a HELOC for the down payment — at the same time?
• Internal bank memos showed employees how to cheat the system to get poor mortgages prospects approved that shouldn't have been: Titled How to Get an "Iffy" loan approved at JPM Chase. (Was circulating that memo also a FNM/FRE/CRA requirement?)
• The four biggest problem areas for housing (by price decreases) are: Phoenix, Arizona; Las Vegas, Nevada; Miami, Florida, and San Diego, California. Explain exactly how these affluent, non-minority regions were impacted by the Community Reinvesment Act ?
• Did the GSEs require banks to not check credit scores? Assets? Income?
• What was it about the CRA or GSEs that mandated fund managers load up on an investment product that was hard to value, thinly traded, and poorly understood
• What was it in the Act that forced banks to make "interest only" loans? Were "Neg Am loans" also part of the legislative requirements also?
• Consider this February 2003 speech by Countrywide CEO Angelo Mozlilo at the American Bankers National Real Estate Conference. He advocated zero down payment mortgages — was that a CRA requirement too, or just a grab for more market share, and bad banking?
The answer to all of the above questions is no, none, and nothing at all.
The CRA is not remotely one of the proximate causes of the current credit crunch, Housing collapse,and mortgage debacle. As I detailed in Barron's, there is plenty of things to be angry at D.C. about — but this ain't one of them.
The root legislative cause of the current was excessive deregulation. From exempting derivatives from regulation to nonfeasance on the Fed's part in supervising bank lending, the pendulum swung too far away from reasonable oversight. By taking the refs off of the field and erroneously expecting market participants could self-regulate, the powers that be in DC gave the players on Wall Street enough rope to hang themselves with — which they promptly did.
There are too many people who are trying to duck responsibility for the current mess, and seeking to place blame elsewhere. I find this to be terribly important, as we seek to repair the damage amidst an economic crisis. Rather than objectively evaluate the present crisis in an attempt to craft an appropriate response, the partisan hacks are trying to obscure the causes of the current situation. Like burglars trying to destroy the surveillance tape, they are all too aware of their role in the present debacle.
Shame on them for their foolishness or cowardice.
Whenever I see a CRA proponent blathering, I have a "Star Trek moment." That's when Captain Kirk proves to some random alien computer that its basic programming is logically inconsistent. It's the AI (artificial intelligence) version of cognitive dissonance. The computer, recognizing the fraud its entire existence was based upon, seeing the futility of its belief system, at least has the dignity to blow itself up. No such luck with the wingnuts, who merely move on to their next piece of spin . . .
Note in the Sources section, we have a few subtopics: "Sources" is what I use to show where data, quotes and charts are from. "Previously" discusses commentary on this subject we have written in the past. "Related" is a good jumping off point for further reading; lastly, Consistently Wrongis where we point out the willfully misleading tripe written by people who should know better, but publish nonsense anyway. In the case where it appears some are trying to mislead the public, the least we can do is call them out.
Originally published at The Big Picture on Oct 2, 2008 and reproduced here with the author's permission.


text:
After being in Taipei for ten days, I somehow feel like I've seen the whole city… I'm positive that's not true but I've certainly seen a lot: Longshan Temple, Taipeo 101, Nanmen Market, a flower and jade market, Chiang Kai-Shek Memorial, a celebration for the Chinese Valentine's Day, a celebration for the Chinese Halloween (yes, in the same week,) the famous Shinlin Night Market, Karaoke (!) and the National Palace Museum, which holds most of the world's Chinese art, saved or stolen depending on your place in the world.
It's really been a whirlwind of experiences so far—being both busy and a little jet-lagged, it's all blended together a bit. My favorite experience so far is a hike some of us took yesterday. 天上山(The Heavenly Mountain) provided a view and adventure that I thought only existed in guidebooks. We were mostly hoping to get in a little exercise and maybe a few good pictures before our first day of classes (tomorrow!) The almost 5 hour round trip hike took us through a Buddhist temple set in the side of the mountain. There, we were offered water to drink, blessed by a Buddha, said to bring us good luck. As we continued on up the Mountain both official plaques and slabs of stone had Chinese carved into them. I remember two rocks carved with the translations of "Change Mountain" and "Blessed Rock." We are lucky to have "ambassadors" (Taiwanese students who will work with us on the language, but have also joined us in our orientation adventures.) They often are able to help with translations, or the simple necessity of understanding directions well. The end of the hike came just as the sun was setting on Taipei.
Traveling to the "must go" sites in Taiwan has been a great way to orient me to the city, get used to the public transportation (which is incredibly clean,) and get to know the other students on my program. But on the last day activities I am itching for Chinese classes to start. I am looking forward to both to a routine, and to start being seriously challenged with this language.


text:
Finland's Minister of Transport and Communications has expressed her country's readiness to promote cooperation with the Islamic Republic of Iran in communications fields and information exchanges.
Anne Berner during a meeting on Tuesday with Iranian Communications Minister Mahmoud Vaezi said that Finland was ready to establish bilateral ties with Iran in all areas of communications.
'We are ready to promote cooperation with Iran in all key areas of communications, including electronic government and network security,' Berner said.
The Finish minister also said that Helsinki is ready to help Iran develop its digital services in healthcare, transportation and education sectors and at the same time assist the country in the fields of information exchanges and mobile services.
Vaezi on his part expressed Iran's readiness for developing cooperation between the two nations in the areas of communications and information technology and urged enhancement of cooperation between the two country's private sectors.
Iran is eager to seize the chance and cooperate with Finland in the communications areas, the Iranian minister said.


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Moose Kid Comics is back with a third issue, and I have another instalment of Big Things Hiding Behind Small Things in there. This time Sir Tiny has been to the opticians for some snazzy eyewear.
You can read the whole issue (plus the previous two and Christmas Special) at www.moosekidcomics.com.


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|Wildflower Will.
|"Welcome to my little patch of garden."
|For one mostly blind old dog, bliss is found somewhere
between the wildflowers and the pumpkin patch.
|Will watching over the pumpkins.
|His squinty-tough guy pose. (The sun was bright and in his eyes.)
|Cosmos and other wildflowers.
|Where we do our best thinking, reading, writing...or nothing at all.
|Chaos & order.


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Swedish Sunday – Deville
After a nice, long holiday weekend, Swedish Sunday is now back. I hope you all enjoyed the holidays, and I hope you're ready for more great music. Today's featured artist is the always heavy Deville.
From their MySpace bio:
When it all came together at the end of 2003 Deville was born after some years of searching. Through a haze of rock, metal and stoner the members have found a way to do something that feels… The line-up was complete when Åkesson came back from Australia and Hambitzer gave up soulless pop and joined the inseparable duo, Andy and Markus. Since the 2004 line-up there have been over 150 gigs and festivals in Sweden, Denmark, Finland, Norway, Germany, the Netherlands, Belgium, Austria, Switzerland, France, Italy and Portugal. Deville has also appeared in radio and national press in these countries. Daredevilrecords released at the end of 2005 a double feature cd lp with Deville and gothenburgers Sergej the Freak called Sergej the Freak meets Deville. Deville signed to Buzzville Records in 2007 and the album Come Heavy Sleep was released in Benelux, December 2007 and in the rest of Europe and the US in the beginning of 2008. Hail the Black Sky is the latest album released in June 2009 again through Buzzville in Europe and in the US and the touring in Europe has continued.
Andreas Bengtsson – vocals, guitar
Martin Hambitzer – guitar
Markus Nilsson – drums
Markus Åkesson – bass
Daredevil Records
- Hands of Mine
- From Below
- Devola
- The Wreck That You Make
- Seven
- Lowride
- Get It Right
- Thorn in My Side
All together a very enjoyable Split…the musicianship varies from very decent to great and the production for both bands are great for the most parts. It might be a bit of a run of the mill release, but it's an above average run from a great mill! – Milan Elkerbout (The Metal Observer)
Buzzville Records
- Intro
- Sunset Capricorn
- Come Heavy Sleep
- Black Dawn
- Deserter
- Stillborn
- Earthburn
- Open Gates
- Into the Smoke
- Sweet Blood
- Far Beyond
- Rise Above
- Outro
While the tell-tale Swedish rock aesthetic is prevalent throughout all 13 tracks, the four-piece manages to push out some heavier than expected moments, as on "Black Dawn" and "Rise Above." Both tracks have a rumbling tone that falls between the syrup of Acid King and the pile-driving mayhem of Black Cobra and are immediate highlights of Come Heavy Sleep. There's a bit of radio sensibility – in both the vocals and the composition – to Deville as well. Come Heavy Sleep does the job of laying down some kick ass stoner rock. Lots of good songs on this one. – John Pegoraro (stonerrock.com)
Buzzville Records
- Levitation
- On the Throne
- Hail the Black Sky
- Undead
- My Enemy
- The Only Thing
- Reason
- Through the Blade
- Early Grave
- Down to Me
While spinning the new Deville album Hail the Black Sky, I had an overwhelming urge to lie down on the carpet and make a (what I like to call) Fuzz Angel. Somewhat similar to a Snow Angel only you flap your arms very fast until you build up enough static electricity to power a small car, then run around screaming "RELEASE THE FUZZ!!!". Typical Saturday night at my house. Anyhow… Hail the Black Sky definitely releases the fuzz, delivering a nonstop barrage of catchy hooks and memorable riffs. – Adam Walsh (planetfuzz.com)


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Welcome to our Team's Homepage
Feb 05, 2010 by Linda Mallut
Welcome to our Light The Night Walk homepage.
In Honor of Becky-Linscott-Houseman my "Honored Hero"
Light The Night Walk is The Leukemia & Lymphoma Society's evening
walk and fundraising event. It is the nation's night to pay tribute and
bring hope to thousands of people battling blood cancers and to
commemorate loved ones lost.
The Leukemia & Lymphoma Society (LLS) funds lifesaving research
that has contributed to major advances in the treatment of blood cancers
and treatments for other types of cancer, such as chemotherapy and stem
cell transplants. These treatments have helped patients live better,
longer lives. New targeted therapies that kill cancer cells without
harming normal tissue are providing drugs and procedures that are
improving quality of life.
- A donation of $25 provides patients and their loved ones with FREE
- booklets that contain up-to-date information on their disease and help
- them make informed decisions about their treatment options.
- A donation of $50 makes possible a Family Support group with a trained
- facilitator where comfort can be found and experiences can be shared among
- patients and family members.
- A donation of $100 helps supply laboratory researchers with supplies
- and materials critical to carrying out their search for cures.
- A donation of $1,000 makes possible one- on-one conversations with
- health care specialists who provide patients with information about their
- disease, treatment options, and helps prepare them with questions for
- their health care team.
Please make a donation to support my team's participation in the Light
The Night Walk and help save lives. Be sure to check my team's Web site
frequently to see my team's progress, and thanks for your support!0


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Budding armchair design experts: I have a new logo to go on the redesigned JAFA website. What colour/s would you use and where to make this logo absolutely perfect? Be sure to look at the colours at http://www.janeaustenfestival.com/ as it needs to work with this.
JAFA is organised under the Earthly Delights banner at http://www.earthlydelights.com.au - Earthly Delights provides an office, insurance coverage, financial backing and me.
12-14 April 2019
Jane Austen Festival Australia is an annual celebration in Canberra where Austen and Napoleonic fans from all over Australia come and indulge themselves in everything Regency - including dancing, music, food, games, archery, fencing, theatre, promenades, grand balls, talks, workshops, costumes and books. This festival is now a regular part of the ACT Heritage Festival, Australian Heritage Week and is supported by the ACT Government, the Australian Costumers Guild and the Earthly Delights Historic Dance Academy. Since its inception in 2008 this little festival has blossomed into one of the most delightful four days anyone could experience each April in an old and beautiful part of Canberra, the Federal capital of Australia.
November 02, 2013
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Tuesday, June 9, 2009
Tabistry Tutorial Now Available at Etsy.com
Well, I've finally gone and done it. I posted my "Tabistry Basics" Tutorial on Etsy.com. In the near future, I will be putting up some of my patterns, as well. I've had so many people requesting to purchase my patterns, I had to get something up. I really hate charging for it. I have been enjoying the fun of swapping with others for crafts. I love all the unique items we have received. I truely cherish each of them. I still plan to offer the swap option, but right now my swaps are kinda getting hard to manage. :)
Hope it goes well and is useful to others. If you do purchase it, I'd love to hear any comments on how I can make it better and as easy to follow as possible.
Hope it goes well and is useful to others. If you do purchase it, I'd love to hear any comments on how I can make it better and as easy to follow as possible.
Posted by mieljolie at 9:29 AM
Labels: announcement
2 comments:
Congratulations, I'm so happy for you and I can't wait to see the patterns you'll be selling!ReplyDelete
That's so sweet of you, crafterella! Thanks! I guess I have to contribute something out of this to my family so they'll let me keep doing it. ;)ReplyDelete
I'm going to try to put some other patterns for nonclothing items up along with a few of my wearables. Anyway, I need a taller trash can for the bathroom with a lid (our IG likes to shread TP). So, guess what's on my list, now?! :)
We've decided that any money made by this venture will go straight to the daughter's college fund. Maybe, someday, when someone asks her how she paid for college, she can tell them, "with can tabs". ;D
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On January 19th, 2013, the unpaid employees of George Soros, Josef Ackerman (Mr Tsipras' new best friend!) and Lloyd Blankfein will be attacking the Hellenic national resurgence. While the Neo-Conservative Samaras destroys the Greek people to pay off amoral loan sharks, what the media calmly has labeled a "Conservative-Marxist" coalition in power in Greece right now is throwing everything in their arsenal against the fast climbing Golden Dawn.
SYRIZA, the spiritual continuation of PASOK, has learned how easy and effective it is to utilize the brutality of some illegal immigrants and young, bored, middle class Anarchists. While SYRIZA tells the Greek people it will deport illegal immigrants to other parts of Europe, they turn around and manipulate the latter into violent attacks on the million strong legal citizens using constitutional rights to free speech and assembly of Golden Dawn. While the media constantly lambasts the Greek people for shutting down and calling for the prosecution of the incredibly vulgar and offensive play called Corpus Christi , the "skeptics" and "free speech" activists and globalist "human rights" NGO's of the West stay remarkably silent when Golden Dawn offices are bombed or lawful assemblies are violently attacked. The new "liberal" idea of free speech and tolerance is the same as Josef Stalin's, the freedom to agree and tolerate the toxic ideology of a tiny clique, no room for anything else! The poster above organized by "tolerant" and "peaceful" leftists is not actually calling for a national exchange of ideas, the useful idiots that come out to these events say it in plain terms : SMASH, CRUSH, DESTROY GOLDEN DAWN!
A list of the groups attending this January 19th attack on the Greek people demonstrates just how far behind the Leftists and international financiers are with the demands of the Greek people. The vast majority of the people have said it time and time again in easy to understand terms: UNINVITED ILLEGALS OUT OF GREECE!
So who is going to this event, here they are http://greecesolidarity.org/?p=611
Let's see:
Panhellenic Federation of Artists and Entertaiment (POTHA), THE GREEK EQUIVALENT OF HOLLYWOOD. HOMOSEXUALS LIVING THE HIGH LIFE IN THE WEALTHY PARTS OF GREECE.
Intellectuals and Artists HOW MANY OF THESE SMUG BOURGEOISIE HAVE TO COMPETE WITH ILLEGAL IMMIGRANT LABOR OR LIVE IN THE IMMIGRANT GANG INFESTED PARTS OF ATHENS?
Theatre "EMPROS",
Initiative "MAVILI" (Artists), MORE REPRESENTATIVES OF THE BABEL TOWER, FAR AWAY FROM THE RAPES, ASSAULTS, AND MURDERS.
Then the list goes on to talk about 1 or 2 vague trade unions, led by more bourgeois college Marxists. What sane craftsman or union worker would want illegal immigrants replacing them for half the pay? Why would they , we could infer by participating in this march, focus so much on Golden Dawn, and not on SYRIZA and New Democracy who are in power and have either started implementing austerity or stood by with calculated idleness to sell them out?
But the best part of all, the bulk of the sponsors? A long list of ethnic organizations, with the praise of SYRIZA, in Greece! :
Union of Immigrant Workers,
Pakistani Community of Greece,
Afghan Community of Greece,
Open School of Immigrants in Piraeus,
Egyptian Community,
Union of United Afghan Refugees in Greece,
Iranian Political Refugees Union,
Antifascist Group of Albanian Immigrants,
Union of Syrians in Greece,
ASANTE (African immigrants in Greece)
So this is the question I propose to those Greeks that are racist against themselves as well as these non-Greek ethnic organizations: Why is it ok for Africans, Albanians, Pakistanis, and Afghans illegally living in Greece to organize based on ethnic lines, but not Greeks… in Greece?
Does the irony escape them? Probably not. They know it's unfair, it's unprincipled, that it's hypocritical for groups that encourage organizing based on race to accuse and stifle another group for doing the same…in their own country!
This is just a macro version of the squatting epidemic, where a Greek that is barely getting by goes to visit the grave of their parents in another town, only to come back and find Anarchist dope fiends or illegal immigrants squatting in their house, with an attitude of entitlement no less! They want to squat in Greece, under a racial pretense that benefits their own race over Greeks, all while condemning the actual native people for "racism" when they loosely try to save their home!
It goes without saying that a Greece for Greeks is not some radical idea invented by the Nazis, it is the default of nature as any human being knows instinctively. As is a Pakistan for Pakistanis, an Afghanistan for Afghanis, an Africa for Africans, all human beings have a right to a home and sovereign government that puts them first. Greece did not create the economic and political conditions in Afghanistan and Africa that immigrants are fleeing, and Greece is in absolutely no position to solve the problems of Africa and Asia either! The people who are today robbing Greece are the ones who bring instability to all these remote regions of the world, these illegal immigrants should go home and fight to fix their own home as the Greek people are trying to do (against the racist replacers, against the communists, against the bankers, and against the traitors). The Leftist (tolerant only of those on their side) and Conservative (Conservers of monopoly capital and usury) alliance has no problem when foreigners within their borders do it, only when the indigenous people of Greece seek to assert their Providence bestowed right to a nation and call on their corrupt government to actually enforce the existing constitutional clause barring all illegal immigration!
Denounce the hegemonic, agenda pushing NGO's, denounce the traitors on the Left and on the Right, denounce "Arab Spring" pseudo-rebels that persecute Christians in Syria and Libya, denounce injustice. Victory to Golden Dawn!


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For answers to common questions, visit our FAQ page.


text: Last weekend, we joined Scott on his Young Adult/College Retreat with Hephzibah Baptist Church (our church). We went to Myrtle Beach and stayed at Kingston Plantation. We had a super nice room with a king size bed that I had to get a running start to get UP onto, and a huge whirlpool tub with jets. We (Briana and Elijah) had to spend a good chunk of time in the room, so I made use of that nice tub!! Elijah and I joined in on the meals and the music, but then left once the speaker started... it would have been a huge distraction to stay with the group.Elijah had a GREAT time in the sand. He would pick up handfulls of sand and let it sift through his hands. After a little while on the beach, we decided it was a lost cause to keep him clean and just went ahead and amused him by buying his legs and feet in the sand. He was quite funny digging his legs back out.We also got in the lazy river, which is like a pool that makes you go in circles so that you can just float with the current. He LOVED that too..This is the group of Young Adults/College Students and Leaders'Coop' took a liking to Elijah.... calling him 'little man'. They had to get a picture together before he could leaveMontage of what Elijah did with the SandDaddy covered me up... pay attention to the toe sticking out :-)Im so happy...Kisses in the sand :-)Stand Up!!Here you go Mommy, have some sand


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Gary Angle
Gary spends his days at O'Brien mired in architectural design and imaging for conceptual and constructed projects. His imaging expertise spans the best of old and new technologies, bringing a unique perspective to the team. Gary's outstanding conceptual representations are a critical tool in facilitating client decision making.
Originally from Fort Worth, Gary received his Bachelor's in Architecture from University of Texas at Arlington. He's married to horticulturist Christina with four grown daughters and three grandsons. Gary is captivated by the beauty of Italy and its people, and has vacationed in Bellagio and Lenno for the past 15 years. He is a bagpipe, drum, and fife fan with plans to attend the Royal Edinburgh Military Tattoo in the near future.


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The Bible says:
Proverbs 22:6 Train up a child in the way he should go; even when he is old he will not depart from it.
My first born
Matthew 7:12 "So whatever you wish that others would do to you, do also to them, for this is the Law and the Prophets.
My Big "D"
Proverbs 29:17 Discipline your son, and he will give you rest; he will give delight to your heart.
My Sweet "D"
James 1:17 Every good gift and every perfect gift is from above
One of my greatest joys
Proverbs 1:8 NIV
Listen, my son, to your father's instruction and do not forsake your mother's teaching
Is 21 years old....
Now a junior in college...
does a parent ever feel like they said enough?
showed enough?
did enough?
for them to remember..
all those things you hope for them
he's back at school
in his first apartment
with all kinds of "hand me down" furniture
leftover camping silverware
and a hodge podge of other treasures to make his place comfortable
he has settled in...
quite perfectly!
We drove up today
to wish him a great day
take him to lunch
and buy him some groceries
There is NO WAY
in this world
that we could be more proud
the man he is becoming.
from what we are seeing...
he DID hear all of the teachings of his childhood
"For whoso findeth me findeth life, and shall obtain favour of the Lord.Proverbs 8:35
We pray prayers of blessing and hope for our Big "D"!!!
We love you more than words can express!!!


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Apologies, my warm and yeasty baker's dozen of dedicated readers, for the suspension of transmission. My mother is here, a watercoloured smudge of Australia, motherly love and faint resentment in her wake. Your regular programming will be back shortly. In the meantime, I wanted to share this t-shirt from snorgtees. It speaks to me.
Women's stories
1 day ago


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By: Marisa Herman Associate Editor
Proposed plans to redevelop part of the city's Old School Square Historic District with a new development known as Midtown Delray will come before commissioners for approval or denial this month.
Originally scheduled to be discussed at a February meeting, attorneys for the developer Delray Beach-based Hudson Holdings asked to postpone the hearing so they could make additional changes to the $140 million project.
Midtown Delray proposes adding a hotel, apartments, restaurants and shops. The 7-acre project is located on the south side of West Atlantic Avenue between South Swinton Avenue and Southwest First Avenue. It is an area that holds historic value to the city, but also an area that needs a facelift as many of the structures are dilapidated.
Attorney Bonnie Miskel called those changes beneficial to the project and said they address concerns residents have raised. The request to postpone will allow city staff time to review the revisions and make a recommendation to the commission on whether they comply with city development rules.
Miskel said the proposed changes will eliminate three of the five waivers the developer is requesting to build the project.
"Our plans will change, not significantly, but positively," she told commissioners.
In a 4-1 vote, commissioners agreed to delay the vote on the project so the developers can submit their updated version of the development for consideration.
Commissioner Shelly Petrolia cast the sole dissenting vote on the postponement.
"This has been going on for a very, very, very long time," she said. "It's been delayed and delayed and here we are again delaying it again. I think we have given plenty of time and I think we should move forward."
The project has gone through several iterations and names like Swinton Commons since it was first proposed several years ago.
Mayor Cary Glickstein said he had concerns about affording the developer due process and supported the delay along with the rest of the commission.
"Our job is to get the best possible project we can," he said.
Commissioners are scheduled to vote on March 6.


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"Absolutely nobody could ever persuade me to do something."
Camila Cabello has denied claims that Taylor Swift was behind her decision to quit Fifth Harmony to pursue a solo career.
Despite speculation at the time of her departure from the group in December 2016, the Havana singer insists that the decision was hers, and hers alone.
"It annoyed me that people said she was encouraging me because I know that she probably got a lot of negative criticism for that", she told The Sun.
"Absolutely nobody could ever persuade me to do something. If I don't want to do something the whole world could be telling me and I won't do it.
"And if I do then the whole world could be telling me to not do it and I'll do it. I've always been that way."
"To be honest with you, she's never ever had anything to do with that. It's so funny, because when Taylor and I talk and whenever we hang out we talk about love and boys."
But while Taylor might not have been behind her decision to leave, she's certainly giving her a boost – after she was announced as a support act for Taylor's forthcoming world tour.
"This is a Reputation stadium tour update, and I've been wanting to tell you this for a really time," said Swift last month.
"The opening acts on the the Reputation Stadium Tour will be Charli XCX and Camila Cabello. I'm really excited, I hope you are too. I can't wait to see you, I can't wait to see them – I'm really excited just about the whole thing in general."


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Using Instagram to document life in Trondheim, Norway, Jan Ove paints a clean and peaceful picture of his hometown. Working as a graphic designer/illustrator, Jan takes a fairly symmetrical approach to his Instagram compositions, which often show a quality mix of both the nature and architecture of Trondheim. Be sure to check out and follow his extremely beautiful feed here @janove. Place : live in Trondheim, Norway. sum up Trondheim? The country's 3rd largest city, and in Norway that means 179,000 people :) In a place like Trondheim everything is pretty calm, safe, clean and a little bit boring. The good kind of boring. Norway was a poor country up until the 70's (oil) so there´s a lot of relics/old stuff from our peasant past which I find interesting and eye pleasing! In small big city like Trondheim and it's surroundings you find a nice mix of urban and rural culture. You have the fjord, the mountains and the stuff in between. I like it. For the visitor? If in Trondheim you should visit the cathedral Nidarosdomen, Stroll around Bakklandet and visit the café Bakklandet Skydsstation, walk along the fjord on the path Ladestien, check out the small wooden houses in Ilsvikøra, take the boat out to Munkholmen and take a trip out of town and check out the small/stange/ugly/pretty places and the nature surrounding the city. Best Meal in the City? Klubb in the restaurant Tavern
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Trondheim, Norway - Jan Ove
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@yinofyang : it is my favorite shot as well, the reflection and the contrast photo, it was good ! :)