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The Epstein-Barr virus (EBV)-encoded lytic activator Zta is a bZIP protein that can stimulate nucleosomal histone acetyltransferase (HAT) activity of the CREB binding protein (CBP) in vitro. We now show that deletion of the CBP bromo- and C/H3 domains eliminates stimulation of nucleosomal HAT activity in vitro and transcriptional coactivation by Zta in transfected cells. In contrast, acetylation of free histones was not affected by the addition of Zta or by deletions in the bromo or C/H3 domain of CBP. Zta stimulated acetylation of oligonucleosomes assembled on supercoiled DNA and dinucleosomes assembled on linear DNA, but Zta-stimulated acetylation was significantly reduced for mononucleosomes. Western blotting and amino-terminal protein sequencing indicated that all lysine residues in the H3 and H4 amino-terminal tails were acetylated by CBP and enhanced by the addition of Zta. Histone acetylation was also dependent upon the Zta basic DNA binding domain, which could not be substituted with the homologous basic region of c-Fos, indicating specificity in the bZIP domain nucleosome binding function. Finally, we show that Zta and CBP colocalize to viral immediate-early promoters in vivo and that overexpression of Zta leads to a robust increase in H3 and H4 acetylation at various regions of the EBV genome in vivo. Furthermore, deletion of the CBP bromodomain reduced stable CBP-Zta complex formation and histone acetylation at Zta-responsive viral promoters in vivo. These results suggest that activator- and bromodomain-dependent targeting to oligonucleosomal chromatin is required for stable promoter-bound complex formation and transcription activity.
Chromatin modification is thought to be an early and integral step in the regulation of transcription and DNA replication (2, 18, 27, 28, 30, 44). Initiation sites for transcription and DNA replication are specified by sequence-specific DNA binding proteins which are subject to multiple levels of regulation through signaling pathways that respond to changes in intra- and extracellular conditions. Higher order chromatin structures associated with repressive heterochromatin are thought to prohibit sequence-specific transcription and DNA replication factors from binding to their recognition sites. Precisely how sequence-specific factors can access their sites in these regions of the genome, recruit chromatin modifying and remodeling activities, and initiate the processes of transcription and DNA replication remains poorly understood (50). Previous work, however, has suggested that a subset of bZIP proteins can stimulate chromatin modifying activity in the absence of sequence-specific recognition, suggesting that these proteins may function as early-acting factors in the chromatin remodeling process (9).
The Epstein-Barr virus (EBV) immediate-early protein Zta is a transcription and DNA replication factor that must disrupt chromatin-repressed latent viral genomes to initiate the lytic cycle transcription program (15, 39; reviewed in references 34 and 43). Zta is a member of the bZIP family with significant sequence similarity to the basic and zipper regions of the adipocyte differentiation factor C/EBP, the erythroid-specific factor NF-E2, and the protooncogene c-Fos. In latently infected B lymphocytes, the EBV genome exists as a nonintegrated nuclear episome with nucleosome phasing indistinguishable from that of cellular chromatin (32, 52). The majority of viral genes required for lytic replication are repressed but can be reactivated by overexpression of Zta or by treatment of cells with pleiotropic agents, such as sodium butyrate, trichostatin A, calcium ionophores, or phorbol esters (34, 43). It is thought that chromatin-based repression is one important component of maintaining the transcription silence of lytic cycle gene products during viral latency. Consistent with this hypothesis is the fact that transcriptional coactivators with histone acetylase activity can cooperate with Zta to stimulate lytic cycle gene expression (1, 55).
The CREB binding protein (CBP) and its close relative, p300, are transcriptional coactivator proteins with intrinsic histone acetyltransferase activity (3, 41; reviewed in references 6, 7, and 26). CBP and p300 are essential for early embryonic development and can costimulate transcription driven by numerous cellular and viral transcription factors (47, 53). The association of CBP and p300 with several activators can be regulated by phosphorylation, and it is thought that recruitment of CBP and/or p300 can be limiting for transcription initiation of several target genes (7, 25, 48). In this way, CBP and p300 are thought to integrate the complex network of transcription signals. The inactivation of p300 and/or CBP by viral oncogenes, including SV40 T antigen, adenovirus E1A, and papilloma virus E7, may play an important role in cellular growth transformation. The near ubiquitous requirement for CBP and/or p300 in transcription regulation underscores the importance of coactivators and histone acetylation in coordinating gene expression. Despite the enormous number of transcription factor interactions with CBP, the precise mechanism of CBP coactivator function remains poorly understood, and it is especially unclear whether CBP can function to facilitate the initial recognition of sequence-specific factors to their cognate binding sites in promoter regulatory regions.
Zta activates transcription of numerous viral and cellular genes in the early stages of the EBV lytic reactivation cycle (10, 19, 33, 37). Zta exists as a homodimer that binds with high affinity to a large family of degenerate sequence recognition sites found in the upstream regulatory regions of responsive viral genes (36). In addition to activation of transcription, Zta also binds to several sites in the EBV origin of lytic DNA replication and recruits virally encoded lytic replication enzymes to the origin (24, 45, 46). Thus, Zta not only is a transcriptional activator but also participates directly in the process of DNA replication initiation. The amino-terminal transcriptional activation domain of Zta has been examined by several groups in some detail (11, 16, 20). It has been found that aromatic hydrophobic residues in the activation domain are essential for transcription activation and recruitment of CBP (16). The Zta transcriptional activation domain binds to the cysteine-histidine (C/H)-rich regions 1 and 3 of CBP, and this interaction is strictly dependent on the aromatic residues that are critical for transcription function (16, 55). It was also found that Zta can stimulate CBP histone acetyltransferase (HAT) activity directed toward small oligonucleosomes (SONs) in vitro (9). Stimulation of HAT activity was also dependent on the aromatic residues in the Zta activation domain, suggesting that a strong correlation exists between the activation of HAT activity and transcription. Stimulation of CBP HAT activity was found to be dependent on the Zta DNA binding domain as well, and this correlated with the ability of Zta to bind directly to oligonucleosomes. Similar concentrations of other bZIP or CBP binding proteins did not share this activity, with the exception of C/EBP and NF-E2, two bZIP proteins with high sequence similarity to the Zta basic DNA binding and zipper dimerization domains. These results suggested that a subset of bZIP proteins shared the ability to direct CBP HAT activity to nucleosomal DNA lacking an obvious sequence recognition site (9).
In this study, we further characterize the activation of CBP HAT activity by Zta and suggest that the mechanism of activation is more complex than the simple recruitment of enzyme to substrate. We found that the activation of EBV lytic transcription correlated with the stimulation of CBP HAT activity by Zta, and both required the CBP bromo- and C/H3 domains. We also found that the basic region of Zta is essential for targeting CBP to oligonucleosomes, which also correlated with transcription activation. Finally, chromatin immunoprecipitation (ChIP) assays revealed that stable promoter-bound CBP and histone acetylation are dependent on the CBP bromodomain. Based on these results, we propose that Zta initiates nucleosomal targeting of CBP HAT activity, which facilitates stable promoter binding and sequence-specific transcription activation.
Cells and plasmids.293 and D98/HR1 cells were passaged in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum and antibiotics. Raji and ZKO-293 cells were passaged in RPMI supplemented with 10% fetal bovine serum and antibiotics. EBV reactivation in Raji cells was induced with 100 ng of tetradecanoyl phorbol acetate (TPA) per ml and 1 μM ionomycin (Sigma). The BRLF1-luciferase plasmid contains nucleotides −178 to +28 relative to the transcription initiation site cloned into the NheI and HindIII sites of pGL3Basic (Promega). BHLF1-Luc has been described previously (16). CMV-CBP-ΔHAT (L1690K/C1691L), CMV-CBP-ΔBromo (Δ1101-1171), and CMV-CBP-ΔC/H3 (D1805-1890) were generated by site-directed mutagenesis of pCMV-CBP-FLAG (gift of M. G. Rosenfeld). Baculovirus expression plasmids for CBP-FL, CBP-ΔBromo, and CBP-ΔHAT were generated by subcloning the CBP gene from the cytomegalovirus (CMV) expression vector construct into the NotI site of pFASTBAC (Pharmingen). Zta wt and ZFZ were expressed in the pQE8 bacterial expression plasmid (Qiagen). ZFZ was derived from a mammalian expression vector provided by G. Miller and consists of Zta (nucleotides 1-171), Fos (nucleotides 137-162), and Zta (nucleotides 198-245) (22). Zta substitution mutants were generated by site-directed mutagenesis (Stratagene Quick Change) in pCDNA3 (Invitrogen) for mammalian cell expression and in the pQE8 (Qiagen) vector for expression in Escherichia coli. Saccharomyces cerevisiae NAP1 was expressed from plasmid pET28-yNAP1 (a gift from T. Tsukiyama). Mammalian expression vectors SRα (pcDLSRα296) and SRα-Zta have been described previously (55). 293 and D98/HR1 cells were transfected with Lipofectamine 2000 (Invitrogen), and luciferase assays were performed as described previously (16).
HAT assays and kinetic analysis.Typical HAT assays contained 200 ng of histone substrate (SONs, dinucleosomes, mononucleosomes, or free histones), 0.25 μCi of [3H]acetyl coenzyme A (Amersham), and 20 ng of baculovirus-expressed CBP in a 30-μl reaction volume as described previously (9). For kinetic studies, baculovirus-derived CBP (20 ng) was mixed with 0.1, 0.2, 0.5, or 1.0 μg of purified HeLa SONs in a 15-μl reaction volume and incubated at 30°C for 1, 2, 5, or 10 min in the presence or absence of 200 ng of Zta. Histone acetylation was quantitated by phosphorimager analysis using a tritium-sensitive screen. Lineweaver-Burk analysis was used to calculate Vmax and Km.
Nucleosome assembly in vitro.Nucleosomes were assembled in vitro by use of yeast NAP1 as an assembly protein as previously published (23), with modifications by T. Tsukiyama (personal communication). Yeast NAP1 was expressed and purified from E. coli by MonoS chromatography. Histone octamers were purified from HeLa nuclear pellets by hydroxyapatite chromatography to remove nucleosomal DNA (14). Chromatin was assembled in a reaction containing 0.1 mg of bovine serum albumin per ml, 0.75 mg of yNAP1, 0.3 mg of histone octamer, and 0.3 mg of pBKSII plasmid DNA in buffer ExB (10 mM HEPES [pH 7.6], 50 mM KCl, 5 mM MgCl2, 0.5 mM EGTA, and 10% glycerol). Nucleosomes were assembled by incubation for 4 h at 30°C. Assembly was confirmed by a gel mobility shift assay (data not shown) and by a supercoiling assay with topoisomerase-relaxed plasmid DNA as described previously (23). Topoisomerase I was purchased from Topogen, Inc. Histone acetylation reactions were described previously (9). Mononucleosomes and dinucleosomes were assembled by high-salt dialysis and purified by glycerol gradient centrifugation as described previously (13).
ChIP.ChIP was performed as described previously (42). EBV sequences were amplified with the following primers: BMRF1 sense, ACCTACATGACTAGCATCAAGCAA; BMRF1 antisense, GGCCTCCATAGTTTACAGACAGAA; BHLF1 sense, AGGTGTGTCATTTTAGCCCG; BHLF1 antisense, CCTACCTCTAGGCTCCACCC; Cp sense, AAAATTGAACCTTGTTGGCG; Cp antisense, ATGGTAAGAACCCTGCGATG; EBER sense, AGGTCAGACCACGAAAGGTC; EBER antisense, AGACAGACTACGTCACCGTG; BZLF1 sense, ATCTTCAGCAAAGATAGCAAAGGT; BZLF1 antisense, TAGAGTCCATGACAGAGGATTTGA; BRLF1 sense, TTCTGCCATGCAATACAAATAAAT; BRLF1 antisense, ATTTCCTCCAGAAGAGTGACATC; cellular gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sense, TCACCACCATGGAGAAGGCT); and GAPDH antisense, GCCATCCACAGTCTTCTGGG. Antibodies for acetylated histones were purchased from Upstate Biotechnologies, Inc. Hemagglutinin (HA) antibodies were obtained from Babco, Inc., and Boehringer. Flag M2 antibody was obtained from Sigma. CBP antibodies (A2) and normal rabbit immunoglobulin G (IgG) were purchased from Santa Cruz Biotechnologies. EBV-specific antibodies for Western blotting were monoclonal anti-Rta (Argene), anti-EA-D (Advanced Biotechnologies), and polyclonal rabbit anti-Zta.
CBP bromo- and C/H3 domains are required for transcriptional coactivation by Zta.We have found that cotransfection of CBP with Zta can cooperatively stimulate transcription from several viral promoters in transient transfection assays (55). In a previous study, it was found that deletion of the CBP HAT domain compromised transcription activation of the BZLF1 promoter, indicating that HAT activity is required for the autoactivation of Zta gene transcription (9). We and others have subsequently found that the CBP HAT domain is particularly important for the coactivation function of the BRLF1 promoter (Fig. 1B) (1). BRLF1 encodes the Rta transactivator, which is required for full lytic cycle gene expression and is a critical early target of Zta transcription regulation. To further explore the mechanism of CBP coactivation, we tested whether deletion of additional subdomains of CBP affected transcription stimulation by Zta. We deleted the CBP bromodomain and E1A interacting region of the C/H3 domain. The bromodomain has been implicated in chromatin (38) and acetyl-lysine binding (17, 29, 31, 54), and the C/H3 domain is the target of numerous protein interactions, including contacts with Zta (55). Consistent with previous findings, full-length CBP modestly stimulated Zta activation of the BRLF1 promoter (Fig. 1B). Cotransfection of CBPΔHAT repressed Zta transcription activation of BRLF1, indicating that HAT activity is important for BRLF1 activation. We now show that deletion of the bromodomain (CBPΔBromo) and the E1A interacting region of the C/H3 domain (CBPΔC/H3) also inhibits transcription activation of BRLF1 by Zta (Fig. 1B). Since Zta can bind to the C/H1 domain of CBP, we suggest that these mutations in CBP function as dominant-negative inhibitors that interfere with Zta and native CBP associations. This suggests that native CBP requires the bromo- and C/H3 domains for coactivation of the BRLF1 promoter by Zta.
CBP full-length and internal deletion mutants were tested for the ability to stimulate viral reactivation from latently infected D98/HR1 cells. Viral reactivation was measured by Western blotting of early antigen D (EA-D). We found that CBPΔHAT and CBPΔBromo were modest inhibitors of Zta activation, while full-length CBP stimulated EA-D activation by Zta. Although CBPΔC/H3 was barely detectable by Western blotting, it had the strongest inhibitory effect on EA-D coactivation by Zta (Fig. 1C). These results are consistent with the findings shown in Fig. 1B and indicate that CBP HAT, bromo- and C/H3 domains are important for Zta coactivation of latent EBV genomes as well as of transiently transfected reporter genes.
The CBP bromo- and C/H3 domains are required for stimulation of nucleosomal HAT activity.Previous work had indicated that Zta stimulated CBP HAT activity directed toward SONs. We have previously shown that HAT activity of immunoprecipitates of CBP expressed in 293 cells could be stimulated by the addition of recombinant Zta in vitro. We also showed that CBPΔHAT lacked HAT activity in immunoprecipitates and could not be further stimulated by the addition of Zta, indicating that the CBP HAT domain and not a CBP-associated HAT was required for HAT activity. We now tested whether the CBP bromodomain or the C/H3 domain was important for mediating Zta stimulation of CBP HAT activity (Fig. 2). Flag-tagged full-length (FL) CBP, CBP-ΔBromo, and CBP-ΔC/H3 were expressed in 293 cells and subjected to immunoprecipitation with anti-Flag antibody (Fig. 2A). As controls, we used baculovirus-expressed and -purified CBP-FL and the transcription factor NF-E2, which had also been shown to stimulated CBP HAT activity in vitro. Addition of Zta or NF-E2 to baculovirus CBP-FL stimulated HAT activity as expected (Fig. 2B). Similarly, Zta and NF-E2 stimulated HAT activity of immunoprecipitates of Flag-tagged CBP-FL. In contrast, neither Zta nor NF-E2 was capable of stimulating CBP-ΔBromo. Interestingly, we found that HAT activity of immunoprecipitates of CBP-ΔC/H3 was not stimulated by Zta and was stimulated at reduced levels by NF-E2. These results suggest that Zta and NF-E2 may have different mechanisms for stimulating CBP HAT activity, although both activators require the bromodomain.
To confirm these results and rule out the possibility that additional cellular factors bound to CBP were influencing the properties of CBP deletion mutants, we assayed CBP deletion mutants that were expressed and purified from baculovirus (Fig. 2C). It was previously found that baculovirus-expressed CBP, but not CBPΔHAT, could be stimulated by Zta (9). As expected, Zta stimulated HAT activity of full-length CBP when SONs were used as substrates. In contrast, CBP-ΔBromo had robust HAT activity when assayed with free histones but had no detectable HAT activity with SONs in the presence or absence of Zta. Deletion of the C/H3 domain also reduced Zta-activated acetylation of SONs but had little effect on free histone acetylation. These results indicate that the CBP bromo- and C/H3 domains are required for Zta-stimulated nucleosome acetylation by CBP, suggesting that this activity correlates with transcription activation in vivo.
Stimulation of CBP HAT requires nucleosome assembly.Histone octamers stripped of nucleosomal DNA by hydroxyapatite were not substrates for Zta activation, while SONs could be stimulated for acetylation (9). To better understand the substrate specificity of SONs, we determined whether in vitro-assembled nucleosomes on supercoiled plasmid DNA could reconstitute the substrate specificity for Zta and CBP. Yeast Nap1 functions as a histone chaperone that deposits histone octamers on supercoiled DNA resembling natural chromatin to some extent (23, 40). Nucleosomes were assembled in vitro using HeLa octamers, yeast NAP1, and supercoiled Bluescript plasmid DNA (Stratagene), which lacks any obvious high-affinity Zta binding sites. Nucleosomes were assembled in the presence of yNap1 (23), and the extent of the assembly reaction was determined by examining the change in superhelical density by use of a topoisomerase relaxation assay (Fig. 3B). We found that nucleosomes assembled in this manner could not be acetylated by CBP, suggesting that nucleosome assembly inhibits histone acetylation by CBP (Fig. 3C). However, the addition of Zta strongly stimulated acetylation of histones assembled in the presence of yNap1. In contrast, octamers incubated with DNA in the absence of yNap1 were not assembled (Fig. 3B), did not inhibit CBP acetylation (Fig. 3C), and were not further acetylated by the addition of Zta (Fig. 3C). These results indicate that Zta can facilitate the acetylation of nucleosomes assembled in vitro on supercoiled DNA. Inclusion of specific Zta-responsive element (ZRE) binding sites in the assembled templates did not significantly alter the stimulation of histone acetylation, suggesting that Zta interaction with nucleosomes was sufficient for this activation (data not shown). These results also indicate that nucleosome assembly inhibits CBP acetylation of histones and that Zta functions to derepress this inhibition.
We also compared the ability of Zta to stimulate histone acetylation on defined-length templates that were assembled by high-salt dialysis and purified by use of a glycerol gradient (13). We found that equivalent concentrations of oligonucleosomes, dinucleosomes, and mononucleosomes were acetylated at similar levels in the absence of Zta (Fig. 3D). However, Zta-dependent HAT activity was significantly higher for SONs and dinucleosomes than for mononucleosomes. These results indicate that oligonucleosomal length is an important parameter for substrate targeting by Zta.
Zta stimulates acetylation of all lysine residues in histones H3 and H4.Specific lysine residues in histones H3 and H4 have been associated with particular functions in gene expression and chromatin assembly in various organisms and developmental models. To determine if Zta targets specific lysine residues for acetylation by CBP, we first utilized acetyl-lysine-specific antibodies in Western blots (Fig. 4A). SONs were assayed in vitro with recombinant CBP and Zta in the presence of unlabeled acetyl-coenzyme A and were subjected to Western blot analysis with various lysine-specific antibodies. Antibodies for the hyperacetylated forms of histone H3 and histone H4 showed significant stimulation of acetylated histones in the presence of Zta. We also found that antibodies specific for histone H4 K5 and K12 caused significantly increased acetylation after the addition of Zta. Histone H4 K8 acetylation was also stimulated by Zta, but to a lesser extent than other lysine residues. The acetylation of lysine residues on histone H3 was examined by Edman degradation of 3H-acetylated histones (Fig. 4B). In this analysis, we found that Zta stimulated acetylation of all lysines, including K4, K9, K14, K18, and K23. Interestingly, the greatest incorporation of the [3H]acetyl group per mole was at K14, K19, and K23, which are residues not typically thought to be subject to significant levels of acetylation. Edman degradation analysis of H4 lysines was not possible since the amino termini of H4 were blocked in HeLa cells. Nevertheless, these results indicate that Zta stimulates CBP acetylation of all lysine residues in the amino-terminal tails of histones H3 and H4 and may preferentially stimulate lysine residues at the C-terminal ends of the N-terminal tails.
The basic DNA binding domain of Zta is required for stimulation of HAT activity.Our previous data suggested that the Zta bZIP domain provided a nucleosome binding function that is important for stimulation of CBP HAT activity (9). Substitution of the Zta basic region with the c-Fos basic region (ZFZ) has been shown by others to allow binding of Zta to AP-1 sites and stimulation of transcription from AP-1-responsive reporter genes to levels similar to those of wild-type Zta. ZFZ failed to activate lytic cycle gene expression since it could not bind to all ZRE sites. However, substitution of serine 186 with alanine resulted in the ability of Zta to bind ZRE sites and to activate EBV lytic promoters in transfected reporter plasmids but in its failure to activate chromatin-associated viral genes (21, 22). To determine if the Zta basic region conferred specific properties important for chromatin-specific reactivation function, we assayed several basic region substitution mutants for the ability to stimulate CBP nucleosomal HAT activity, bind nucleosomes and ZRE DNA, and stimulate transcription from plasmid or viral chromosomal targets (Fig. 5). Substitution mutants S186A, S186E, VASR-MAAA, AKF-NKR, QLLQ-ELLD, and ZFZ (Fig. 5A) were expressed and purified from E. coli, normalized by Coomassie staining, and assayed for the ability to stimulate acetylation of SONs by CBP (Fig. 5B). We found that S186E, MAAA, and ZFZ had significantly reduced stimulation of HAT activity. These same mutants were then assayed for nucleosome binding by an electrophoretic mobility shift assay (EMSA) with 32P-labeled SONs (Fig. 5C) (9). We found a perfect correlation between nucleosome binding and stimulation of HAT activity, suggesting that nucleosome binding is necessary for HAT stimulation by Zta. These substitution mutants were then assayed for binding to a ZRE from the BHLF1 promoter (Fig. 5D). We found that all of the substitution mutants, except for ZFZ, were capable of binding to this ZRE, indicating that nucleosomal binding was distinct from DNA binding. These same mutants were then assayed in transient transfection assays for activation of BHLF1 promoter fused to luciferase. Transcriptional activation of the transfected plasmid DNA correlated perfectly with the ability of Zta mutants to bind nucleosomes and stimulate CBP HAT activity (Fig. 5E). However, when these mutants were assayed for the ability to activate transcription from viral chromosomal genes, we found that S186A, in addition to S186E, MAAA, and ZFZ, was defective (Fig. 5F). This result is consistent with the findings of Francis et al. (21, 22), who showed that S186A is specifically defective for transcription of viral chromosome-associated genes. These findings with S186A suggest that stimulation of CBP HAT is not sufficient for transcriptional activation of viral chromosomal genes and that posttranslational modification of this residue may also be required for viral reactivation in vivo (4). Nevertheless, these data suggest that nucleosomal binding is a specific feature of the Zta basic region, is not found in the c-Fos basic region, and is essential for stimulation of CBP HAT activity.
The mechanism by which Zta stimulates CBP HAT activity was further analyzed by use of Michaelis-Menten kinetics (Table 1). Purified baculovirus-expressed CBP was assayed in the presence or absence of Zta with SONs (ranging from 0.1 to 1 μg) to generate plots for 1 to 30 min of reaction time. Lineweaver-Burk analysis was used to calculate the Km and Vmax for three independent experiments. We found that the addition of Zta decreased the Km by an order of magnitude but had only an approximately twofold effect on Vmax. This suggests that the predominant mechanism of activation of Zta is to increase the effective local concentration of nucleosomes for HAT catalysis by CBP.
Zta stimulates histone acetylation in vivo.Latently infected Raji Burkitt lymphoma cells can be induced to enter the lytic cycle by treatment with TPA and ionomycin, which stimulates transcription of the BZLF1 promoter (49). This treatment has been shown by others to lead to hyperacetylation of the BRLF1 immediate-early promoters (8). However, in these studies it was not clear if TPA and ionomycin treatment involved Zta or CBP for gene activation and histone acetylation. We now show that TPA-ionomycin leads to an increased association of Zta and CBP with immediate-early promoters BZLF1 and BRLF1 in vivo by using a ChIP assay (Fig. 6). We also found that TPA-ionomycin leads to an increase in hyperacetylated histones H4 and H3 at the BZLF1 and BRLF1 promoter (Fig. 6). This result demonstrates that CBP functions in association with Zta at the immediate-early promoters during in vivo reactivation, and this correlates with an increase in histone acetylation.
We have shown that Zta can stimulate CBP histone acetylation in vitro and transcription activation in vivo. To determine if Zta stimulates histone acetylation in vivo, we assayed histone acetylation by ChIP after transfection of latently infected D98/HR1 cells with a Zta expression plasmid (+) or control vector (−) (Fig. 7). Since Zta stimulates viral replication in these cell lines, input viral copy number was increased three- to ninefold after transfection with Zta (Fig. 7, input). ChIP revealed that histone H3 and H4 acetylation increased at EBV lytic promoters but not at a viral latent transcript (EBERS) or a cellular gene (glutaraldehyde-3-phosphate dehydrogenase [GAPDH]). Quantitation of the ChIP PCR products as percentages of input DNA revealed that lytic immediate-early promoters BZLF1 and BRLF1 were increased two- to fourfold with acetylated histones H3 and H4 (Fig. 7B). In contrast, Zta transfection did not stimulate histone acetylation at EBERS or GAPDH, and no PCR product was detectable with control IgG antibody (Fig. 7). These results indicate that Zta can stimulate acetylation of histones H3 and H4 at transcriptionally responsive lytic viral gene promoters.
The ChIP assay was next used to determine if the bromodomain of CBP was important for histone acetylation at EBV-responsive promoters in vivo (Fig. 8). Zta was cotransfected with the CBP-Flag or CBP-ΔBromo-Flag expression vector into highly transfectable ZKO-293 cells. As expected, the addition of Zta increased the input copy number for EBV DNA. Interestingly, the addition of CBP, but not CBP-ΔBromo, further increased input DNA levels, suggesting that CBP stimulates DNA replication. ChIP assays with antibodies to Zta, Flag (CBP), and acetylated histones H3 and H4 revealed a striking increase at EBV lytic promoters (BRLF1, BMRF1, and BHLF1) relative to latent transcripts (EBERS and Cp) or cellular genes (GAPDH). By adjusting PCR levels as percentages of input DNA to normalize for replicating genomes, we found a consistent two- to fourfold increase of Zta, CBP, and acetylated histones at EBV lytic promoters. Importantly, this increase was dependent upon the bromodomain, since transfection of CBP-ΔBromo had no significant effect on histone acetylation or promoter association of Zta or CBP. CBP and CBP-ΔBromo were expressed at similar levels and did not affect Zta expression levels, as determined by Western blotting (Fig. 8B). These results indicate that the CBP bromodomain is required for the increase in histone acetylation at Zta-responsive promoters and for the stable association of Zta and CBP with promoter DNA in vivo.
Histone acetylation is thought to be a prerequisite for transcriptional activation of genes that may be silenced by heritable chromatin structures (5, 28). Transcriptional coactivators with intrinsic HAT activity can be recruited by sequence-specific DNA binding proteins to alter the local histone acetylation pattern and to stimulate transcription complex assembly (28, 51). The mechanism by which HAT complexes facilitate sequence-specific DNA binding of transcription factors remains poorly understood, since it is not clear how the initial sequence recognition occurs in higher-order chromatin structures that limit DNA accessibility. One simple mechanism that would overcome this problem is for initiating activators to recruit HAT complexes to nucleosome arrays found in repressed chromatin. Our data suggest that the EBV immediate-early bZIP protein Zta stimulates CBP acetylation of nucleosomal chromatin to enhance transcription complex assembly at viral lytic gene promoters.
Previously, it was shown that a class of bZIP proteins which included Zta, C/EBP-β, and NF-E2 was capable of stimulating CBP HAT activity directed towards oligonucleosomes (9). Here we further characterized the ability of Zta to stimulate CBP HAT activity. Most significantly, we found that the CBP bromo- and C/H3 domains were important for Zta activation of transcription and nucleosomal HAT activity. The bromodomain was required for both Zta and NF-E2 stimulation, but the C/H3 domain deletion had a greater effect on Zta activation, suggesting that Zta and NF-E2 may stimulate HAT activity by different mechanisms. Zta binds to the C/H1 and C/H3 domains of CBP with near equal affinity in vitro (55). Deletion of the C/H3 domain may reduce CBP binding to Zta, but it is unlikely to eliminate the strong interaction with C/H1 and therefore is unlikely to eliminate CBP-Zta association in vivo. We suggest that Zta binding to two domains of CBP provides allosteric enhancement of nucleosome-specific histone acetylation. Kinetic studies showed that Zta had a weak but reproducible effect on the Vmax of CBP HAT activity, suggesting that some stimulation of catalysis may exist (Table 1). However, the predominant effect of Zta was on the Km, which was reduced ∼10-fold in the presence of Zta. This suggests that Zta alters the substrate accessibility, either by recruitment of CBP to nucleosomes or by altering substrate-enzyme interactions to make acetylation more favorable. A similar observation was made with RpAp48, a histone binding factor that stimulates CBP HAT domain targeting to core histones and nucleosomes (56). Although simple recruitment of CBP to nucleosomes can stimulate nucleosome acetylation, we consider it more likely that Zta facilitates the accessibility of nucleosomal histone tails to the catalytic HAT domain of CBP. However, direct evidence for any conformational change in CBP or in the histone tails after the addition of Zta awaits further experimentation.
Bromodomains from several different proteins have been shown to interact with acetylated histones, and it has been proposed that the bromodomain four-helix bundles bind acetylated lysine (17, 31, 54). The p300 bromodomain, which is most closely related to the CBP bromodomain, was shown to be important for transcription initiation and responsiveness to estrogen receptor activation in vitro (35). The p300 bromodomain was also found to bind to chromatin and was essential for a stable, template-committed transcription complex (38). The bromodomains of Swi2 and GCN5 were also found to be required for anchoring of these chromatin-modifying complexes to acetylated chromatin templates (29). Binding to acetylated lysine may be important for the recognition of nucleosomal substrates, since we also observed that CBPΔBromo had a reduced ability to acetylate oligonucleosomes relative to free histones (Fig. 2). We suggest that the bromodomain facilitates processive targeting of multiple lysine residues in the amino-terminal tails of all four histones. Bromodomain association with acetylated lysines may help target adjacent lysine residues on neighboring histone tails. We hypothesize that Zta facilitates this process by interactions with the C/H1 and C/H3 domains of CBP as well as by providing essential interactions with nucleosomes.
Several lines of evidence suggest that the bZIP domain of Zta interacts with nucleosomes to facilitate histone acetylation by CBP. Zta bound HeLa-derived oligonucleosomes in vitro and stimulated histone acetylation of nucleosomes assembled on defined plasmid templates lacking known high-affinity ZRE sites (Fig. 3). Nucleosome assembly in vitro inhibited CBP acetylation of core histones, which could be reversed by the addition of Zta, suggesting that Zta altered the accessibility of histone tails in packed nucleosomes. Substitution of the Zta basic region with the c-Fos basic region eliminates the ability to target SONs for acetylation. Mutation of a single amino acid residue in the Zta bZIP domain from serine to glutamic acid inhibited nucleosome binding and HAT activation but did not eliminate ZRE DNA binding (Fig. 5). This suggests that S186 is important for nucleosome-specific binding and HAT activation. Interestingly, mutation of Zta serine 186 to alanine had no effect on HAT activity but eliminated transcription activation of viral chromosomal, but not plasmid-borne, genes. The complex behavior of S186 mutations suggests that additional events, like phosphorylation, may regulate the nucleosome binding and activation properties of Zta in vivo (4).
ChIP assays with TPA-ionomycin-induced Raji cells or Zta-transfected cells indicated that histone acetylation was increased at specific viral lytic promoters but not at viral latent transcripts or at cellular genes. Zta is known to stimulate transcription from these viral lytic genes (16), indicating that a good correlation exists between transcription activation and histone acetylation of Zta-responsive promoters. Interestingly, we found that cotransfection of wild-type CBP, but not CBP-ΔBromo, increased histone acetylation of Zta-responsive genes, as well as increased the promoter association of Zta and CBP themselves. Thus, transcription activation is correlated with increased occupancy of the activator-coactivator complex. This in vivo observation is consistent with recent biochemical studies showing that the bromodomain anchors coactivators to nucleosomal templates (29). The bromodomain-dependent increase in Zta binding to Zta-responsive genes suggests that CBP and Zta function in concert to select specific promoter sequences in nucleosome-repressed chromatin.
Higher-order chromatin structures can present a significant barrier to transcription initiation and DNA recognition by sequence-specific DNA binding proteins. Nucleosome binding and chromatin opening activity have been described for the "pioneering" proteins HNF3 and GATA4, which initiate liver-specific gene expression during development (12). Zta may also function as a pioneering protein with a different mechanism of action. Our data suggest that Zta recruits CBP to compact chromatin to induce transient histone acetylation, which in turn facilitates scanning for high-affinity Zta binding sites in compact chromatin. This pioneering function may be shared by other bZIP proteins, like C/EBP-β and NF-E2, that have a similar ability to target the nucleosomal HAT activity of CBP (16). The results presented here demonstrate the importance of the bromodomain as a nucleosome targeting element that may facilitate processive acetylation of histones in compact chromatin and promoter scanning by sequence-specific activators such as Zta. Finding the structural basis through which Zta facilitates nucleosome acetylation in a bromo- and C/H3 domain-dependent manner remains the work of future studies.
We thank W. Wunner and the Vector Expression Core at the Wistar Institute for baculovirus infections and acknowledge the technical support of Latasha Day.
This work was supported by a grant from NIH (CA-85678).
- Received 19 November 2002.
- Accepted 13 January 2003.
- American Society for Microbiology


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Come and join us on Sundays and Wednesdays at our Los Banos Campus. Located at 935 I St, Los Banos, CA 93635. Bring the entire family and see how there is something for everyone at Lifetree.
9:30am fellowship
Come and join us for FREE coffee and baked goodies prior to our 10:00AM service. We believe fellowship is vital to our growth, so bring the entire family and we look forward to welcoming you to the Lifetree Family!
10:00am kid's church
Ages 5-12 years of age are invited to participate in King's Kids every Sunday morning! It is our prayer that as we teach the children about the heroes of the Bible, they will discover how they can become a hero for God themselves.
10:00am service
"Let the word of Christ dwell in you richly, teaching and admonishing one another in all wisdom, singing psalms and hymns and spiritual songs, with thankfulness in your hearts to God." Colossians 3:16 ESV / 142
WEDNESDAY NIGHTS 6:00PM Los Banos
Our Wild 'N' Out youth group meets every Wednesday night at 6:00PM right here in Los Banos. Come for a great evening of fellowship with food, video games and prizes. All Jr. High & High School students are welcome!

subset: commoncrawl


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Yesterday I received my first package from ASOS and I was excited to wear this mint green dress! I'm so happy I decided to get it. The material is really thick and a nice quality... plus the color is so BRIGHT! I love it!! There are two ways I was thinking about wearing it: either in a chic way or a more boho way. I went with chic by accessorizing with this studded collar plus a gold studded bracelet and gold necklace. Lavender socks and a coral belt gave the outfit an extra funkiness. =)
Love this dress so so much! Hey! I was on this website > wonderrocket.com/ec/cmShopTopPage1.html
Do they ship worldwide? I don't speak this language but I would be so happy to puchase to them! Thanks for answere!


text: 20th Anniversary and Awards of the League of Maritime Editors & Publishers in photographs 11/02/2019 admin Uncategorized 0 L-r: DR. Kofi Mbiah, former MD Ghana Shippers Authority, presents to Barr. Nicholas Bruce, the Life Achievement Award for the Growth and Development of Nigeria's Maritime Industry bestowed on Mr. Pier Luigi Carrodano, Group Managing Director, NALComet Group L-r: Dr. Kofi Mbiah presents to Mr. Adeniyi Olayiwola Shittu the award of Innovation in Customs Brokerage in Nigeria given to his father Prince Olayiwola Shittu, the immediate past President of the Association of Nigeria Licensed Customs Agents. Mr. Muyiwa Akande, Manager, Corporate Affairs, SIFAX Group, receiving the award of Best Indigenous Terminal of the Year given to Ports & Cargo Handling Services Ltd from Barrister Hassan Bello, Executive Secretary of Nigerian Shipper Council Mr. Adams Jatto, General Manager, Public Communication & Strategy (right) receiving the Award of Transparency and Infrastructural Development in Nigerian Ports given to the NPA from the Executive Secretary of Nigerian Shippers Council, Barrister Hassan Bello. L-r: Otunba Kunle Folarin, Mazi Tony Ohakwe, Mr. Kingsley Anaroke, President of League of Maritime Editors & Publishers,Prince Olayiwola Shittu and Mr. Wale Oni. Share this:Click to share on Twitter (Opens in new window)Click to share on Facebook (Opens in new window)Click to share on Google+ (Opens in new window) Related


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It is always a good thing to book a travel insurance a short while before you take holiday in Mount Desert Island.
If you have planned holiday to Mount Desert Island in United States of America you would probably visit Somesville and Northeast Harbor, also Seal Cove in Town of Tremont. During your journey in Mount Desert Island, United States of America you will have the opportunity to enjoy strolling in public gardens, for the more adventurous, there is kayaking and even carrying wallet in backpocket.
Currency there is Us dollar and the general consumer price index is high. Generally you will find the health-care cost bordering to high, and Mount Desert Island Hospital is known as safe. Last thing to know, criminality rate is high so with all this information choose your travel insurance carefully.


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Companies in Croydon Park, Australia
Looking for Companies in Croydon Park? Here you will find 407 companies in Croydon Park, Australia. For more results, browse business categories or keywords.
Categories in Croydon Park
- Beauty Professionals
- Construction
- Engineering
- Advertising
- Beauty Salons
- Logistics
- Doctors and Clinics
- Clothing and Accessories
- Advertising Agencies
- Furniture Manufacturers
Top Cities
We found 407 companies
RM Gregory Promoprint
227-231 Georges River Road,, Croydon Park, NSW
R.M Gregory Printers is a leading printing business firm that has garnered multiple awards & accolades through the years. We strive to offer our high quality products and services in a friendly & loya...
VerifiedPhoneE-mailWebsitePhotos (4)
336 South Road, 5008, Croydon Park, SA
Ozaboriginal Pty Ltd
222 Georges River Road, 2133, Croydon Park, NSW
Altec Group Pty Ltd
23 Hay St, 2133, Croydon Park, NSW
Altec Group PTY. LTD. Pty Ltd : Printed Circut Boards and Quick-Turn Prototypes, Sydney, Australia
Cavan Glass Pty Ltd
26 Rawson Street, 2133, Croydon Park, NSW1
ANZ ATM Croydon Park
BP Express 236-240 Georges River Road, 2133, Croydon Park, NSW
ANZ offers a range of personal accounts and services including internet banking, credit cards, personal loans, insurance, home loans and foreign exchange. Find out more now.
39846 ALISON ST, 2133, Croydon Park
16 DUNMORE ST CROYDON PARK, Croydon Park
A A AI.L G M Spares
35-37 Beresford Avenue, 2133, Croydon Park, NSW
ABA Cement Rendering Pty Ltd
23 Lennartz St, 2133, Croydon Park, NSW
230 GEORGES RIVER RD, 2133, Croydon Park
Adelaide Industrial Labour Service Pty Ltd
Corner of South and Overland Roads, 5008, Croydon Park, SA
AFU Pty Ltd
PO Box 397, 2133, Croydon Park, NSW
All Tech Electrics
PO Box 320, 2133, Croydon Park, NSW
A Stratos Electronics
NSW 2133, Croydon Park, NSW
Battery World Northern Metro
151 Regency Road, 5008, Croydon Park, SA
Black Forest Bakery
32 Days Road, 5008, Croydon Park, SA
Buckley Pty Ltd
19 Brighton Avenue, 2133, Croydon Park, NSW
WORKS DEPOT KINGSBURY ST, 2133, Croydon Park
PROVINCIALATE 1/ 2 ALISON ST, 2133, Croydon Park
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Age: 17Location: BaliSchool: Junior High SchoolOrphan: NoTEP Number: IBBnTM-135
Karang's parents broke up in 2012. His father left town and his mother could not support him on her salary. He lives in a one-bedroom house with his paternal grandmother. He is a funny, friendly child who always wants to help out at the center. He wants to be a doctor when he grows up.
To support I Putu Aditya (Karang), please see the options below:


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1378 results found for ""
Kitchen Confidential
[mustHaves] Liven up your counter with ChicWrap, the stick tight, easy-to-use cling wrap, that lets you say goodbye to grated knuckles thanks to their single finger slide button. Available in six energetic designs. www.bonvivant.ky [image issue="Fall...
October 04, 2014
Delectable Dishes
Cayman Style Sauce [mustHaves] 8 oz canned peeled tomato 4 oz whole peeled tomato 4 oz yellow onion 2 oz red pepper 2 oz green pepper 1 oz garlic 2 oz Pickapeppa Sauce 1 oz butter 1 tsp scotch bonnet Salt and pepper to taste [image issue="Fall_2014"...
October 04, 2014
Killer Cocktails
[mustHaves] Ketel Part:1 Featuring distinct citrus flavours not expected from a clear martini, this zesty elixir is a delightful surprise. 2 oz Ketel One Vodka 1/2 oz homemade orange syrup 1/4 oz Grand Marnier Pinch of ascorbic acid 2 drops orange bi...
October 04, 2014
Mirai - The Future of Green Homes
Set to break ground in 2015, Mirai is the latest vision from the team of eco-experts at GreenTech Group, a Cayman Islands based conglomerate providing sustainable products, services and eco-expertise. [image issue="Fall_2014" article="mirai" image="m...
October 04, 2014
Contemporary Cayman
Since the late 2000s, interior design and architecture has taken a modern turn in Grand Cayman with a number of high-end developments breaking ground. Similarly, buyers and renters are increasingly requesting contemporary fixtures and finishes, as we...
October 04, 2014
Designing New Horizons
OBM International Founded in Bermuda almost 80 years ago, OBM International has, in a sense, gone forth and conquered the world, architecturally speaking. On that journey they have gained wisdom, experience and international acclaim. Now they are ref...
October 04, 2014


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Step 1
Select a legal name for your partnership and perform a name availability search using the search function on the website of your state's Secretary of State. Some states require the legal name of a partnership to be the last names of the partners -- for example, "Cramer & Cramer."
Step 2
Select a fictitious business name, perform a name availability search and register your fictitious name, if you wish your partnership to be known under a name other than your last names. Many states require you to notify the state government of your fictitious business name, and others allow you to register a fictitious business name even though it is not mandatory. Registration will prevent another in-state business from using your name.
Step 3
Draft a partnership agreement in duplicate, sign both copies and have your spouse sign them. You don't have to file your partnership agreement with any state or federal agency. The partnership agreement should cover your respective capital contributions to the partnership, profit and loss sharing, voting rights, management rights and responsibilities, entry and exit of partners and dispute resolution procedures. Because a two-person partnership faces a tie vote every time the partners disagree, a comprehensive partnership agreement is the most effective way to resolve disagreements.
Step 4
Amend any title documents representing property that you want the partnership to own so that it lists the legal name of the partnership as the owner. If the partnership will own real estate, for example, have the county land recorder's office reissue the title deed. Use a quitclaim deed to relieve the previous owner of potential liability in case of unrecorded mortgages or competing claims to title.
Step 5
Register as a limited partnership with your state government, if you want one partner to enjoy limited liability for partnership debts. To register you must complete a short registration form and pay the state filing fee. The registration form asks for the name and address of the partnership, the name and address of a legal representative, and the name and address of the general partner. Designate either yourself or your spouse as the limited partner in the registration form. The limited partner may not take an active role in operating the business.
Step 6
Apply for any business licenses necessary to do business in your state or locality. A real estate partnership, for example, may require state licensing of the partnership itself, even if the partners are already licensed realtors.


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Call us @ 561.744.3232
for a free estimate
As a leading provider of murphy beds, we take pride in offering the best craftsmanship available in the Jupiter and surrounding areas.
We are dedicated to serving the needs of our customers each and every day.
We have been in business for over 30 years.
Our customers are always happy to provide references.
Look for us on Angie's list!


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Snack Review
Buy Herr's Snacks on Amazon #ad
Note: This snack has been discontinued by the manufacturer.
Taste test: These wide-rippled potato chips had a very sweet flavor on them, about as sweet as you'll ever get on potato chips.
The seasoning seemed to be a combination of mustard, onion and various other spices. The taste wasn't entirely unlike honey, mustard and onion chips, but the flavor was even sweeter in this case, with the mustard more subtle. All of the chips in the bag were small. Pretty good.
Shop: Buy Herr's Snacks on Amazon #ad
Smell test: Nice smell, sort of a sweet honey mustard.
From the package: "The chip with the Sweet Island Flavor" ... "Contains wheat and soy ingredients."
Online store: Buy Herr's Snacks on Amazon #ad
Company: Herr's
Reviews of similar snacks:
This snack was discovered at Snaxpo. Review published .
Most popular in previous months
Still hungry? Check out the 100 latest reviews


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If you haven't heard about the Vivoactive 3, then you're either not a fitness addict, or you're simply not current. In this Garmin Vivoactive 3 review, you'll get to know everything there is about this innovative fitness watch, and why you might consider getting it for your sporting activities.
Like most of the other smartwatches we've seen recently, Garmin Vivoactive 3 is aiming to be more than just a fitness device. The Fitbit series is a perfect example of this, having completely moved to the smartwatch category with the Fitbit Ionic – Apple equally followed the same route, starting as a smartwatch that's since transformed more and more into fitness.
In maintaining Garmin's sports tech legacy, the Vivoactive 3 comes with GPS tracking, heart rate sensor, and a collection of inbuilt apps dedicated to keeping checks on a variety of sports activities, from skiing and snowboarding to running and indoor cycling.
Even though Garmin Vivoactive 3's key focus may be on fitness, this all-round timepiece equally has an ample of other features to make noise about, including real-time stress and sleep tracking, built-in payment system known as Garmin Pay, and also smart notifications.
Garmin has often been a famous name in the fitness and sports industry, so much so that it was able to crush TomTom. And with the Vivoactive 3, the company is pushing vigorously to ensure that this a wristwatch you'll wear daily, which also translates to a new, improved design that's a world away from the popular Garmin Vivoactive HR.
Aside from that, it's equally a Garmin device that's suitable for people who don't wish to go all-in on the Garmin Fenix 5. Basically, Garmin Vivoactive 3 is more affordable and has a lot of the same features, but just removed a few things here and there.
Its design equally reflects its sporty yet smart appearance, as it looks more like a wristwatch than a large fitness tracker. That being said, here's more about the Garmin Vivoactive 3 smartwatch:
- Garmin Vivoactive 3 Review | Design & Highlights
- Garmin Vivoactive Review | GPS and Heart Rate
- Vivoactive 3 Review | Fitness and Sports
- Garmin Vivoactive 3 | App and Smart Features
- Garmin Vivoactive 3 Activity Tracking
- Garmin Vivoactive 3 Accuracy
- Easy to Use Garmin Pay
- App Store | Limited to Fitness
- Garmin Vivoactive 3 | Battery Life
- Garmin Vivoactive 3 Review: Conclusion
Garmin Vivoactive 3 Review | Design & Highlights
Garmin vívoactive 3 GPS Smartwatch – Black & Gunmetal
With such a big jump in design quality, Garmin Vivoactive 3 slightly upped the Vivoactive HR even before I'd turned it on. The unappealing rectangle has been changed by a circular face, whilst keeping the size slightly over 43mm.
That makes the watch 1mm bigger than Garmin Fenix 5S, the smallest of the Fenix series, but a great deal more tiny than the popular Fenix 5 and, of course, the colossus that is the Garmin Fenix 5X. It also weighs only 43 grams, a fraction less than the Garmin Fitbit Ionic, another timepiece it's fiercely competing with.
But as well as weight, Garmin Vivoactive 3 equally loses some buttons, so you'll be doing a bunch of screen tapping and swiping as a result. Although, there's just one button on the right-hand side. All the other Garmin Fenix 5 smartwatches have 5 buttons and transitioning to just one does feel like a significant relegation when Garmin's interface hasn't been immensely improved in turn.
However, Garmin has included a touchpad along the left-hand side of the wristwatch case, which you can scroll by running a finger along it, and it's a good alternative to the touchscreen for moist fingers. That being said, I wish it was a bit easier to navigate the Vivoactive 3, though I assume most of my frustrations with the watch interface can be attributed to the delay it has sometimes when I'm moving between faces.
Furthermore, Garmin Vivoactive 3 comes in 3 basic versions. There's a black colour with a silver ring around its face, but it can be bought with a silver ring and white strap, or one that's all black in colour. The Vivoactive 3 is certainly not the most fashion-conscious sports watch out there, but I'm sure a lot of people will prefer the look over Garmin Fitbit Ionic. I understand that some of you don't just like a square face.
Talking of, the 240 x 240-pixel display on Garmin Vivoactive 3, it's far from the most vibrant I've ever seen and pales tremendously when compared with the Apple Watch and Fitbit Ionic. What I do admire a lot though is the always-on-dimmed display, so I'll never have to shake my wrist to wake it up, as is always the case with Fitbit Ionic. Although, a raise of your wrist will ignite the backlight, for when you need the included visibility.
Another edge Garmin Vivoactive 3 has over the Ionic in terms of design its support for 20mm quick-release straps, this means that it can be paired it with an ample of other straps if the one in the box isn't good for you, or you simply love having other options.
Generally, I'd put the Garmin Vivoactive 3 on a par with the Garmin Fenix 5S in terms of looks and appearance, but above the other Fenix 5 smartwatches and their exposed screws. However, it's like a more efficient Forerunner on your wrist in some ways, or a bit less efficient Fenix 5, depending on how you view it.
Garmin Vivoactive Review | GPS and Heart Rate
Source: wareable.com
We've seen optical heart rate make continuous improvements since the Garmin Vivoactive HR. Although it's not perfect yet, it's gradually getting better at maintaining increased intensity. With the Vivoactive HR, we discovered that it battled pushing above 165bpm, but the Garmin Vivoactive 3 has shown to be much better. In fact, it equally maintained the pace with the Polar H10 chest strap in troughs and peaks, which is a good news for people that run on an interval.
According to the guys at wareable.com:
Furthermore, I equally compared the Vivoactive 3 to the Fitbit Ionic, not only because they're shooting for a similar user, but because Fitbit Ionic happens to be one of the most efficient GPS devices around at the moment. Fitbit made a great deal of this before it was introduced, and it was right. Thankfully Garmin performs commendably here as well, as expected from a brand that has GPS running through its veins.
Fitbit Ionic came in with about 0.05 miles more distance ahead of the Garmin, and there were equally a couple of instances the Garmin seemed to fumble slightly, but nothing to be too worried about. Aside from that, it was equally very quick to get a lock.
Vivoactive 3 Review | Fitness and Sports
Source: wareable.com
Notwithstanding the new smart features, fitness and sports is still the soul of Garmin Vivoactive 3, and the new smartwatch comes fortified with an ample of workouts to choose from. You can equally decide which among these you desire to keep as your favourites – I have swimming and running– but from elliptical to skiing to indoor and outdoor rowing, there's a lot here for everyone.
However, if you don't like any of them, you also have an option to create your own, where you can simply rename an existing activity, altering it so you have the alerts, data screens, and sensors working just the way you like it. But there are chances that you'll be set with what Garmin has packed on there.
A single press of that side button will instantly start any activity, which will move you straight to your favourites. Interestingly, the process of starting an activity is pretty quick – much faster than the Fenix Ionic and probably the Apple Watch as well depending on how you set it up. The moment you've got a GPS lock – or not, assuming you don't need it – it's simply another tap on the button to begin, and you'll be notified by a vibration to get moving.
It's really average for the course, but Garmin Vivoactive 3 builds on the Vivoactive HR with an iota of new features. Basically, it can monitor VO2 Max now, a measurement of the highest saturation of oxygen in the course of the exercise, and a vital element in measuring your fitness. Because of that, we've seen it creep into Apple's and Fitbit's devices, and Garmin pioneered it with their Vivosmart 3 earlier. I don't believe this is better on the Vivoactive 3 because of the heart rate sensor demonstrating a bit more accuracy.
But apart from that, there's equally stress monitoring, and that also a feature that Garmin introduced with the newest Vivosmart, which further embolden the Vivoactive 3 as an all-day timepiece. This works by tracking the variability of your heart rate all-through the day and accessing your stress levels.
Garmin Vivoactive 3 equally allows you to download and use structured workouts either from the mobile app or the desktop Connect platform, and start them from the wrist. This can be done when you start an activity and tap the workouts button, which will reveal to you all the ones you have synced. This is an amazing feature the Vivoactive 3 has over Fitbit Ionic (at least, until it gets more apps).
Moreover, another upper hand is allowing you to pair a chest band or other Bluetooth devices with the Garmin Vivoactive 3, a feature that's not supported by the Fitbit Ionic.
However, Apple and Fitbit watch outperformed on onboard music storage. Although, I'm the kind of person that tends to take their phone along while on a run, I've been tempted more and more by the idea of going without it. But I stream pretty much, and if you're like me then there's a possibility that you don't have many music files anyway.
But overall, I like changing up my playlists each time I'm out for a run, so streaming from my smartphone makes a lot of sense, but that might change, however, when Apple Music streaming is launched to Apple smartwatch. The Vivoactive 3 can function properly without the smartphone, but if you really want music, you'll be required to keep compressing that phone to your arm, or slipping it inside your pocket before you go.
Although I've already mentioned stress as one of the things Garmin Vivoactive 3 takes care of, it's also tracking your resting heart rate, calories, steps, and active minutes; Garmin obviously wants this device to be an all-day wristwatch. It equally tracks sleep, however, I don't really find sleep tracking feature of Garmin to be as good as other similar devices.
I can confidently say that Fitbit currently holds the gold standard for wrist-based devices. Although Garmin got it right a couple of times, but more constantly missed the mark – and in many cases over – or massively undershot.
The problem with it is that it either assumes I've slept off when I've been immovable for too long (I'm just being lazy, and not sleeping!) or it takes a lot of time to register when I've come out of bed. This could be much better if the heart rate is integrated into Garmin's algorithms, as this is what made Fitbit get a better idea of when I'm sleeping and when I'm fully awake, whilst Garmin is simply making use of accelerometer data (although there are apps you can get that utilizes both).
Garmin Vivoactive 3 | App and Smart Features
So, how much of a smartwatch exactly is Garmin Vivoactive 3? First, let's begin with notifications – any notifications that enter your smartphone, you can have it displayed on the wristwatch. So everything from texts messages to Snaps, to Facebook updates. You can't speak through Garmin Vivoactive 3 though, but you can answer or dismiss phone calls on your smartphone through the watch.
Aside from that, you can equally read complete messages and reply to them using preset replies – but this only works on Android devices. While carrying out my tests, I swapped from iPhone to Android, and instantly I could see that I could respond to text messages with things like "Love you!", "Ok."
Although I don't think I've ever used cut-out replies to this day except when I was testing watches, as it just feels strangely dishonest. But if that's not a problem of concern to you, so long as you're using an Android device you can do it with Garmin Vivoactive 3.
Source: wareable.com
Talking about the new app. The entire smartwatch team has lamented about the Garmin Connect mobile app for a very long time, but to my utmost surprise, I was able to test a beta of the new and enhanced version when it was introduced, and for your information, it is really new and improved.
What's primarily changed here is the home screen, which now displays your daily stats on a series of cards. Once you scroll down, you'll see heart rate, steps, weight, sleep, stress, intensity minutes, and a lot more; tap any of these and you'll be taken to a page with a more detailed breakdown. Additionally, you'll find a summary box of your last 7-days at the very bottom, and you can also manually include any activities or weight changes.
It's very similar to Fitbit's app experience that equally breaks down your day into a visually consumable form – and we love this a lot. And hopefully, it will move out of beta pretty soon because this has been needed for a terribly long time now.
Garmin Vivoactive 3 Activity Tracking
The Vivoactive 3 is sited just below the more sport-focused watches in Garmin's line up like the Forerunner and Fenix series, and so it's no surprise to see them focusing more on sleep, steps, tracking weather, intensity minutes, flights of stairs, and so on.
There is equally a new stress stat that basically tells you how your day is going in numeric form.
With inbuilt GPS, compass, GLONASS, accelerometer, barometric altimeter, Garmin's Elevate wrist-based HR, thermometer, and the Vivoactive tops the majority of activity trackers out there.
There are equally a million different activity modes to select from such as ski, yoga, strength, and even the default bike and run. But apart from that, you can equally create your own or simply get a prebuilt app from the Connect IQ store. This is another part where Garmin has flopped on smartphone integration, and it's unfortunate you cannot define these from inside the app.
The wristwatch can show up to 3 screens of 4 metrics as well as an HR gauge. If you choose the 4 field layout, the outside 2 are limited to only a time of day, distance, heart rate, timer, and calories, while the inner two get the complete spectrum of data fields – the 3 field layout equally permits full customizability for each field.
As expected, you equally have options for auto-scroll, auto-pause, auto-laps and manual laps.
Essentially, the timepiece can connect to both speed and cadence sensors, and ANT+ and Bluetooth HR straps, but it does not natively allow connections to power meters. But there are some data fields in the Garmin Connect IQ store that do let the devices communicate, even though they equally yield for some wonky readings occasionally.
Garmin Vivoactive 3 Accuracy
Source: tomsguide.com
I actually tested it against a Garmin ANT+ heart rate band that's paired to the head unit of a Wahoo Elemnt Bolt and went by the best procedure for wrist-based optical HR – worn just above the wrist bone and not so tight.
The Vivoactive tracked really much similar to the chest strap while on the road. I was so astonished at how close the Vivoactive tracked.
But unfortunately, when it came to the trail, the Vivoactive surrendered to the bumps quite significantly and really battled to retain a lock on my heart rate. Additionally, to ensure that this was not a one-off, I took another MTB ride which showed to be a little better, but still displayed a difference in the result.
On the other hand, the GPS accuracy is bang on – with the distances lining up to around .1 of per mile and average speed around 1mph. This was making use of the normal GPS setting and was even closer to the GPS +GLONASS.
Interestingly a feature I never expected to find on an activity tracker of this quality was the ability to download training and workouts plans from the Garmin Connect app and also have the wristwatch guide you through them.
Easy to Use Garmin Pay
Source: tomsguide.com
Just like Apple and Fitbit, Garmin wants to make its watches capable of doing a lot of things on their own. The brand launched a mobile payment system with the Vivoactive 3 named "Garmin Pay." This payment system lets you pay for items on the go by just holding your watch close to a payment terminal. You can easily tell whether a restaurant or store allows mobile payments if you see an NFC icon in the checkout section.
Interestingly, it's pretty easy and straightforward to set up Garmin Pay in the Garmin Connect phone app. You can manually add payment details or simply snap a photo of your physical credit card, and then verify your credit card with the credit card issuer or bank. The entire process takes just some minutes, but it's well worth it.
Just long press Garmin Vivoactive 3's side button and click on the wallet icon to make use of Garmin Pay in a store. You can also store several cards in your wallet and select between them by simply swiping down. Once that is done, you'll immediately see an image of your card and its last 4-digits to ensure that you choose the correct one.
However, Garmin Pay does not really support a lot of cards at launch, which sadly limits its usefulness. They currently support Visa cards from Bank of America, First Tech Federal Credit Union, Capital One and U.S. Bank and Mastercards from BECU, Capital One, Bank of America, Security Service Federal Credit Union, VACU, and U.S. Bank. But the good thing is that more and more card issuers are being included in the already, but the roster is nowhere close as costly as Apple Pay, which has a long head start.
App Store | Limited to Fitness
Similar to Apple and Fitbit, Garmin also has an app store for its smartwatch, as I already stated earlier. But while Garmin claims that its Connect IQ store has over 1,600 apps, big-time applications that aren't related to fitness are few and scarce. There are an ample of workout apps and watch faces that can be installed from the store in the Garmin Connect app, but they're awfully specific.
The store has apps for skiing, hiking, biking, stretching, running, and setting a stopwatch, but you'll find it quite challenging to find famous smartphone apps that are not related to working out. Typically, an app that displays your Uber driver's ETA and another that does the same for Lyft are incredibly useful, but your smartphone is still needed to hail a ride.
Finally, it's a nice thing that Garmin is so devoted to fitness, but a smartwatch should be more flexible. Garmin Vivoactive 3 would be handier when you're not working out if it had more apps which can deliver news headlines to your wrist, control smart-home devices, and use the watch's sensors to provide more insights into your health. It has a few though, but not enough. And when speaking of fitness, though, Garmin's store is equipped with everything you could want – and even some.
Garmin Vivoactive 3 | Battery Life
Excitingly, that puts it at about 3 extra days ahead of the Fitbit Ionic. However Fitbit Ionic is equally running a much higher resolution display, and possibly streaming music as well. But Garmin has neither of these features, but it sure does have other interesting stuff to its bow that may as well make it a more enchanting choice for you.
Here's a short video review of the Vivoactive 3:
- A very easy to use touch interface
- Extended battery life
- Good aerobic and daily heart rate tracking
- Smart notifications with calendar
- All-inclusive stats in Garmin app
What We Don't Like
- Sleep tracking is not accurate
- Heart rate tracking usually gets confused by intervals
- Messaging apps don't work properly
- Garmin app UI is a mess
Garmin Vivoactive 3 Review: Conclusion
Generally, Garmin Vivoactive 3 is one of the first-class sport-oriented activity trackers you can ever found in the market. Even though it does not have certain smartwatch features like the ability to connect to Bluetooth headphones and memory for music – these are features you can see in the likes of Apple and Fitbit – but the optical heart rate proved to be more accurate and the range of inbuilt sensors make up for this obvious fact.
It features the typical Garmin habit like the app integration and notifications could be better, but the Vivoactive is really only limited by its firmware at this point. Good enough, it has all the internal sensors which come with the more costly smartwatches, it just needs to be informed how to use them.
That brings us to the end of this Garmin Vivoactive 3 review. I'm sure by now, you already know everything you need to know about this smartwatch that will further inform your buying decision. However, if there's anything you really want to find out which we didn't cover on this post, use the contact form to reach us, or simply give us a shout-out in the comment section.
And if you enjoyed this post, please help us spread the word by sharing it with your friends.


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After his triumphant return to the fold last week, a clip from tomorrow's Agents of S.H.I.E.L.D. brings Fitz back to Simmons. This season of Agents of S.H.I.E.L.D. took quite a gamble by leaving out one of its main characters for three episodes, but the reveal that Fitz made his way to the future and space was worth the wait. Even better, last week's episode focused entirely on the absent agent's journey and finally brought Lance Hunter back to the show. Hopefully, Hunter will return when the agents head back to Earth (and their own time), but first they'll need to be rescued by Fitz.
Tomorrow's episode, 'Marauder Protocol,' will see Phil Coulson himself step behind the camera as Clark Gregg directs a mini-Marvel movie for the first time. Aside from Fitz's return, we'll see Daisy forced to battle for the Kree in a gladiator-like battle. Meanwhile, the rest of the team will attempt to make contact with what remains of the Earth's surface. But in pure character terms, it's hard to argue that the reunion of Fitz and Simmons won't be the episode's most emotional moment.
RELATED: Agents of S.H.I.E.L.D. Drops An EPIC Star Wars Reference
Marvel Entertainment released a new clip from this week's Agents of S.H.I.E.L.D. that sees Fitz and his new alien friend Enoch milling around Kasius' gathering of prospective Inhuman buyers. And while the two prove their comedic pairing immediately, Fitz soon becomes distracted when he spots Simmons in her role as one of Kasius' servants.
Likely, what will follow is Fitz attempting to speak with Simmons only to realize she can't hear him given the metallic goo Kasius implanted in her. The moment will no doubt still be dramatic, however. Even the clip itself teems with tension (in fact, there's already fan fiction based on the promo). Though the pair have been separated by hundreds of thousands of miles and 74 years, it's only been 6 months for Fitz. Likely, it's been even less time for the agents in space. Regardless, a lot has kept Fitz and Simmons apart in the past and it's always nice to see them back together.
With Agents of S.H.I.E.L.D. taking over after the limited run of Inhumans, audiences won't have to wait longer in between episodes. The winter break only lasted a week, and it's likely the show will run straight through the fall season until its finale. In the midst of that will be the series' 100th episode, recently commemorated with new Agents of S.H.I.E.L.D. art looking back on the show's major moments. We've heard something special is coming for episode 100, and anything from a big new Marvel Comics character to a return to Earth (and, perhaps, of Bobbi and Hunter) could be in the future. For now, fans will probably be satisfied just to see the team reunited.
MORE: Marvel's Agents of S.H.I.E.L.D. Just Answered A Lot of Big Season 5 Questions
Agents of S.H.I.E.L.D. returns tomorrow, January 5, on ABC.
Source: Marvel Entertainment


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Denia Caballero takes discus WL with 70.65m (231′ 9″)
Last Saturday the Cuban discus thrower Denia Caballero pushed Sandra Perkovic from the top of the world at the XV International Meeting Villa del Bilbalo in Spain. With a massive throw of 70.65m Denia improved her personal best by more than one meter and lacked only 23 centimeters for the North American record of her compatriot Hilda Ramos of 1992. Previously Olympic, World and European Champion Sandra Perovic (Croatia) had led with 70.08m the world's top list. Yaimí Pérez placed second with also a solid distance of 65.11m.
How will discus queen Sandra Perkovic answer to that massive new world leading throw?


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The hardest part for a candidate when selecting their vice presidential candidate is finding the right balance. You want to select someone who adds new demographics of supporters and increases your odds of winning the general election without overshadowing yourself. The nominee doesn't want to select a candidate who is not noteworthy enough to make a difference (think William E. Miller in 1964), different or controversial enough to cause a loss of focus on the candidate's platform (think Sarah Palin in 2008), controlling enough to affect the candidate's superiority as president once they take office (think John C. Calhoun from 1824-1828), or someone with a dangerous personal agenda that could come out later once in power (think Aaron Burr in 1800). Finding that perfect running mate is often a very difficult feat to accomplish. Mitt Romney would know, he's facing this challenge right now.
You would think someone like Romney would be easy to balance off. His only political work was being governor of Massachusetts from 2003-2007 and is better know as a businessman. Romney is also lacking a compelling and powerful personality when it comes to public speaking and debating. So, for balance reasons his running mate selection should be a veteran politician with multiple terms of experience and an excellent orator and debater while still not failing any of the four points made above. Names are constantly being thrown around by the media lately trying to determine a candidate that fits this criteria. The problem is, every single name they have mentioned fails to do so perfectly.
Two of the most popular names as of late are Florida Senator Marco Rubio and Louisiana Governor Bobby Jindal. Both are younger and minorities that could these demographics so naturally they seem like a good selection. The problem with Sen. Rubio is that he could easily overpower Romney's duller personality in such a way that he garners more focus and damages the campaign. Meanwhile, Gov. Jindal for the most part is a political unknown on the national scale and isn't likely to make a difference.
Then there is Paul Ryan, a young Tea Party member who would increase Romney's chances in the primarily Democrat state of Wisconsin. However, Ryan's controversial and somewhat unpopular budget plan would gain attention over Romney and would be the focal point of any debates.
Several other candidates such as Ohio Senator Rob Portman, Virginia Governor Bob McDonnell, and New Jersey Governor Chris Christie are also considered front runners. Sen. Portman makes sense as a safe choice from a battleground state that is a must win. The problem with Sen. Portman is that he is also not a compelling man, does not add any new demographics to the campaign besides Ohio, and was budget director during the debt building Bush Presidency. Gov. Bob McDonnell makes little sense for Romney as well. He would cause a large focus on the controversial ultrasound abortion law he supported this year. This would likely push female voters even further from selecting Romney. Gov. Chris Christie is likely the best available choice for Romney. Gov. Christie has a powerful personality and managed to win as a Republican in a mostly Democrat state, New Jersey. The only apparent flaws for Christie is that he could also overshadow Romney, like Rubio, and that his large weight creates health concerns.
The fact of the matter is the current Republican landscape lacks any candidate (yes, even Ron Paul) who is anywhere near the perfect choice, and this could greatly hurt Romney come November. Did I miss someone that you feel is a perfect or near perfect selection? Or do you wish to refute one of my statements about a candidate not fitting the bill?


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ATLANTA — During a recent taping of "Family Feud," a woman gave an answer that no one could have expected.
Visibly embarrassed, the woman immediately followed her answer with, "I didn't say that," but it was too late.
The audience and her opponent couldn't contain their laughter as host Steve Harvey just stood for a moment in stunned silence.
Needless to say, the answer wasn't on the board.
Watch the video above to see the hilarity ensue.
33.748995 -84.387982


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Electric Guitar - Mexican Stratocaster
Electric Guitar - Mexican Stratocaster
This is an Electric Fender Mexican Stratocaster Guitar. It is in excellent condition and has very good strings. Its pickups are in excellent condition, and the pickup dial is also in very good condition. This guitar stays in tune very well and sounds great.
Add To Cart


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This Wednesday April 17th, 2013 at 10:00am PST ….. I am returning to Soul Revolution Radio. Yes I have been invited back ~ Three times a charm!… Read more "Dream Speak with Soul Revolution Radio"
Category: Interviews
Lette's Chat … about Dreams
On Wednesday Feb. 27th, 2013 I will be returning to Lette's Chat to discuss what is really happening during our dreamtime with host Carlette. We will explore… Read more "Lette's Chat … about Dreams"
Ringing in the New Year ~ Jan 2, 2013 on Walking With Spirit Radio Show
This blog posting was from 2012 Merry Solstice! Peaceful Holidays! and a Happy New Year to all! Now that the darkest days of the year are passing,… Read more "Ringing in the New Year ~ Jan 2, 2013 on Walking With Spirit Radio Show"
Thursday, 12/20 on The Soul's Journey with Tom Jacobs : Unraveling Karma
Other Radio Shows to download ~ click here *************************************** I wanted to share with everyone the following blog posting (from Astrologer Tom Jacobs website). Hope you can… Read more "Thursday, 12/20 on The Soul's Journey with Tom Jacobs : Unraveling Karma"
My highschool encounter with a familiar
Jacob's Beloved Books invited me to write a guest post. As I write through inspiration, it is sometimes hard for me to write on a time limit.… Read more "My highschool encounter with a familiar"
Follow My Blog Tour!
My book DreamWorkers Behind the Veil; Night Shift Dreams is available for purchase at Oaklight Publishing, Amazon, Barnes & Noble… or support your local bookstore by asking… Read more "Follow My Blog Tour!"
Simple Ways to Cleanse Yourself of Negative or Unwanted Energies
After speaking with David Schrader and Tim Dennis on Darkness Radio last night. I decided to take my list of Simple Ways to Cleanse Yourself that I… Read more "Simple Ways to Cleanse Yourself of Negative or Unwanted Energies"
Darkness Radio Guest – Dreams, Spirits, Healing
Live (10pm to Midnight CST) last night on Oct 2, 2012 I was given the opportunity to be a guest on Darkness Radio I had a lovely… Read more "Darkness Radio Guest – Dreams, Spirits, Healing"
Excerpt from DreamWorkers Behind the Veil; Night Shift Dreams Book
Find My Book: DreamWorkers Behind the Veil; Night Shift Dreams at Barnes & Noble, Amazon, and on Oaklight Publishing website ! Behind the Veil of Night Shift… Read more "Excerpt from DreamWorkers Behind the Veil; Night Shift Dreams Book"
Interview with Kim Larocque from Muse In The Valley
I was lucky enough to be interviewed by Kim Larocque (fellow Dream Intuitive) from Muse In The Valley. Funny, Kim mentions discovering my blog through Zementa (which… Read more "Interview with Kim Larocque from Muse In The Valley"
STARTING TODAY THURSDAY OCTOBER 14TH, 2010 From 11AM TO NOON Flash of Spirit a wisdom based talk show about dreams, spirituality, an messages sent to us in… Read more "FLASH OF SPIRIT RADIO SHOW…."


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- Matella – My self written engine based on modern OpenGL using LWJGL
- OrangeJuice – First attempt at writing a first person based RTS
- Kirella – Taking all experience about first person RTS games to a new project, based on Matella
Yes, I like RTS games. 🙂
Here you can find some of the projects I've been doing in my freetime over the past years. Lot's of them are games (although I don't wanna be a games engineer) as no other area let's you achieve something you can actually see and that easily play around with.
I also got pretty interested into 3D rendering using modern OpenGL as can be seen from my self written engine called Matella.
The past years I've drifted somewhat away from this and have mainly focused on more technical and software engineering wise complicated projects.
I've been contributing to the Forged Alliance Forever open source project since December 2016. FAF basically is a community made Lobby/Tournament/Map-/Mod-Vault project for the real time strategy (RTS) game Forged Alliance from 2007.
I've spent quite some time on one of their clients (Downlord's FAF Client).
The game internally uses a peer to peer network architecture that tends to disconnect players from the game regularly. Due to this I've also invested some time into fixing/writing and ICE adapter lately which aims at tunneling all game connections through the local adapter and using the Interactive Connectivity Establishment (RFC 5245, candidate discovery, STUN+TURN) protocol for connecting to other players.


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Prestonwood's own 500-voice choir joined the fun, adding background vocals to the weekend's stellar lineup which included the Gaither Vocal Band, the Booth Brothers, the Isaacs, Lynda Randle, the Nelons, Karen Peck & New River, Charlotte Ritchie, Buddy Greene, Angela Primm and more.
In a recent Homecoming Magazine article, Bill Gaither reflected on his choice of Texas as the locale for this annual event. "Historically Texas has been a hot spot for gospel music, dating all the way back to the Stamps Music days in Dallas. A lot of people may consider Texas 'the Wild West' and a bit independent, but a lot of great art has originated in Texas, including a number of beloved Gaither Vocal Band members, including Guy Penrod, Mark Lowry and David Phelps. So we have a lot of friends and a lot of history in this great state, and they always come out with Texas-sized enthusiasm."
With gorgeous weather all weekend, two grand evenings of uplifting music, and Homecoming family and friends all around, this year's event truly was … well, spectacular! If you weren't able to make it this year, we hope you'll join in the fun next year!
Enjoy these photos from the Texas Homecoming Spectacular:


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As a B Corp, we believe in harnessing the power of business to help build a more sustainable community. In our journey, we have been influenced by liked-minded parties who aspire to improve the world we live in. In today's post, we share the second part of our conversation with George McGraw, Founder and Executive Director of DIGDEEP, —with whom we love to collaborate on initiatives that move towards a future with clean running water for everyone.
What are some of the future projects/campaigns Dig Deep is working on?
Dig Deep has two kinds of projects:
- Water projects, where we partner with local communities to develop resources for people who don't have clean running water at home.
- And, education projects for people that have water, but often take it for granted like the #4Liters Challenge.
After the #4Liters Challenge this October, we are launching a really exciting project in November, where we would be able to bring water and electricity into homes in the Navajo Nation, New Mexico by installing solar panels, sinks, water heaters, showers and batteries that are water pumped.
Why does DigDeep like to partner with Falcon Waterfree Technologies?
We think the future of conservation is all about taking something that we already do and making it better. It's not about radically disrupting or changing our lives it's about making them friendlier and more efficient, and Falcon understands this. After all, we all have to go to the bathroom, right? So, why not do it in the most sustainable way possible.
It is the perfect example of a small but impactful change that we all can make in our buildings, stadiums and cities.
Plus, we also think Falcon is a young and innovative company, full of dreamers and believers like us, and when we partner in a project, awesome things happen.
Upgrade your commercial building restroom and start saving water and money TODAY with our products. Check our latest PROMO and get four free cartridges with installation of your Hybrid or Waterfree urinals.


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Open floor...
3 comments:
- theoddbod said...
what's your off-season training routine? ps. do you wanna have a home run derby at centennial?
- October 5, 2009 at 8:20 PM
Similar question to the one in a previous comment: Is your off season training tailored to what the Jays' staff said you NEED to work on or is it basically 'Get better and be ready on your own for Spring Training'?
Also, are you still under contract with the Jays?
Enjoyed watching you in N.H. this season and see some good potential there!
- October 6, 2009 at 11:39 AM
I love your posts about what life was like living with your fellow pitchers this season. I'm curious now, what was life like on the road? Living in hotels, transportation to road games in places far away like Akron, if you have free time and what you do in it?
Also, what is it like having basically a revolving door at the bullpen with guys going up and coming down? Do you keep in touch with guys like Zep and Bubbie or is it more of a temporary bond you all share?
- October 7, 2009 at 5:45 PM


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The Mystery of Giulia Farnese Revealed
I wake up to an inbox full of press releases every morning as part of my work editing Where Rome magazine. To be perfectly honest, the majority of them go straight in the trash after a quick skim—either they aren't of interest to my readers, or the event in question is already out of date (I write up to two months out, so I need lots of advance notice). The releases that I do find pertinent get filed away for future reference. Rarely do I ever receive a release that makes me gasp out loud. But it happened a few months ago.
{I wish I could say, "But it happened today." Sounds much more dramatic. But as you have probably realized, I took a loooooooong break from blogging, and am just now getting back into it. So when the release in question arrived, I filed it away for the day I revived my dormant blog. And that day is today, I am happy to say.}
So after that long preamble, what event is it that got me so hot and bothered? Well, perhaps some of my most faithful bloglings remember the post I did on The Borgia Pope, Pinturicchio, and Giulia Farnese way back in 2012. I'm a little obsessed with the Borgias, and I'm very obsessed with art history mysteries. So when I heard that the Capitoline Museums were displaying a rare work by Pinturicchio called Baby Jesus of the Hands (which is actually a fragment of a larger work that is now lost), I was thoroughly intrigued. Check out that post for the full story on the fascinating mystery of that controversial (and eventually mutilated) work of art.
When, five years later, the very same Capitoline Museums announced in the above-mentioned press release that not only was the elusive Baby Jesus of the Hands going on display once again, but that it would be presented alongside Pietro Fachetti's rarely seen copy of Pinturicchio's full work, The Divine Investiture of Alexander VI, giving much needed context to the beautiful Baby Jesus of the Hands fragment, it was all I could do to contain my excitement. The exhibit includes works of art depicting some of the Borgia clan's most infamous members such as the ruthless Cesare, the sensuous Lucrezia, the courtesan-turned-matriarch Vannozza Cattanei, the papa cattivo himself Rodrigo, and another fragment of Pinturicchio's original work, containing the face of Giulia herself. Also on display are additional works by Pinturicchio, a number of ancient Roman sculptures, Renaissance maps and engravings that shed light on what Rome looked like during the time of the Borgias, and even an original copy of Giorgio Vasari's The Lives of the Artists.
The exhibit is still on, dear readers and fellow art lovers, but not for long. So if the story of the Borgia Pope, his lovely young mistress Giulia Farnese, and the painting that immortalized them and their baby daughter Laura is as intriguing to you as it is to me, I suggest you get yourself to the Capitoline Museums post haste. Pinturicchio, Painter of the Borgias: The Mystery of Giulia Farnese Revealed is only on until 10 September 2017!
Below is a selection of the works on display.
All images provided courtesy of Roma Capitale - Sovrintendenza Capitolina ai Beni Culturali-Assesorato alla Crescita Culturale.


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Birmingham (BHX) to Larnaka (LCA)
One Way/Economy
London (LTN) to Larnaka (LCA)
One Way/Economy
*Prices have been available for one way within last 48 hours and may not be currently available. Additional baggage fees for other optional service may apply. Other terms and conditions may apply.
With a history dating back 6,000 years, Larnaca is Cyprus's oldest city. Known for its palm-lined promenades and Mediterranean beaches, it's best seen by foot. Book your flight to Larnaca with Blue Air and see the crossroads of the Greek, Roman, and Ottoman empires.
Tour the Church of St. Lazarus. Discover the baroque iconostasis and the tomb of the biblical Lazarus. Then bike around the Larnaca Salt Lake and see the flamingos on your way to Hala Sultan Tekke Mosque. This revered mosque is an ancient monument and Umm Haram's burial place.
Take a stroll down Palm Trees Promenade and enjoy sweeping views of the Mediterranean Sea. You can hear a concert at the Seafront Stage or learn more about Larnaca's culture at Europe Square and the Larnaca Medieval Castle, both located at opposite ends of the promenade. Then, stop by a cafe for some fresh seafood and people-watching.
Browse the shops along Ermes and Ermou Streets in downtown Larnaca. You'll find exquisite laces and silverware from nearby Lefkara along with independent jewelers and watchmakers.