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Precise regulatory mechanisms are required to appropriately modulate the cellular levels of transcription factors controlling cell fate decisions during blood cell development. Here, we show that miR-126 is a novel physiological regulator of the proto-oncogene c-myb during definitive hematopoiesis. We show that knockdown of miR-126 results in increased c-Myb levels and promotes erythropoiesis at the expense of thrombopoiesis in vivo. We further provide evidence that specification of thrombocyte versus erythrocyte cell lineages is altered by the concerted activities of the miRNAs miR-126 and miR-150. Both microRNAs are required but not sufficient individually to precisely regulate the cell fate decision between erythroid and megakaryocytic lineages during definitive hematopoiesis in vivo. These results support the notion that microRNAs not only act to provide precision to developmental programs but also are essential determinants in the control of variable potential functions of a single gene during hematopoiesis.
The determination of hematopoietic lineages heavily relies upon regulation at the transcriptional level, and the functional roles of many transcription factors that are essential for hematopoietic specification have been well established. 1–4 Throughout hematopoietic development, the cellular levels of these transcription factors undergo dynamic changes that affect lineage specification as well as the proliferation and maturation of a given cell type 5. The c-Myb protein is a major hematopoietic transcription factor whose levels must be precisely controlled during normal blood cell development and whose aberrant expression contributes to leukemogenesis. 6–10 C-Myb functions at multiple stages of blood cell development from hematopoietic stem cells (HSCs) and progenitor cells to fully differentiated cell types, and can affect the definitive specification of lymphoid, myeloid, erythroid and megakaryocyte lineages. Given the spatio-temporal diversity of its developmental roles, the mechanisms controlling the timing and levels of c-Myb activity must be precise and tightly maintained. While the c-Myb protein has a very short half-life, the regulatory control of its degradation, synthesis and mRNA stability remains poorly understood.
MicroRNAs (miRNAs) provide an elegant mechanism that affords finely tuned control of protein levels by controlling translational efficacy and mRNA stability 11. These noncoding small RNA molecules are quickly becoming recognized as key regulators of diverse cellular functions, including the specification and differentiation of hematopoietic cells 12. The c-myb 3′UTR contains predicted target sites for more than a dozen miRNAs 13–15. Up to date, regulation of c-Myb by two of these miRNAs has been experimentally validated. The overexpression of miR-15 inhibits c-myb 3′UTR-dependent luciferase activity in vitro and contributes to the inhibition of erythroid and myeloid colony formation 16. Overexpression studies of miR-150 in vitro, using a luciferase reporter gene containing the c-myb 3′UTR, also result in a reduction in luciferase activity. miR-150 deficient mice show an increase in B1 cells which are known to require c-Myb 17; an increase in c-Myb protein level was also seen in miR-150 deficient activated B cells. These findings support the role of miR-150/c-myb interactions in B-cell maintenance and differentiation 18. Gain- and loss-of-function analyses have shown that the miR-150/c-myb interaction is also essential for the lineage decision of megakaryocyte-erythrocyte progenitor (MEP) cells to form either megakaryocytes or erythrocytes in vitro 19. Overall, these observations indicate that variations in the levels of c-Myb may be differentially controlled by multiple miRNAs to execute lineage–specific developmental programs at critical junctions during hematopoiesis. Nevertheless, possible combinatorial effects of multiple miRNAs regulating c-Myb within the same developmental context have yet to be addressed experimentally.
Zebrafish and mammalian hematopoiesis are highly conserved on molecular and cellular levels and the phases corresponding to mammalian embryonic and fetal hematopoiesis occur rapidly in the zebrafish with definitive hematopoiesis predominating by 5 days post-fertilization (dpf). Therefore zebrafish provide an ideal system to study the function of miRNAs on different cell lineages at specific hematopoietic stages 20. Furthermore, zebrafish are amenable to transient gene knockdown assays using antisense morpholinos and forced mRNA expression analyses, which provide major advantages for studying mechanisms of miRNA activities regulating embryonic development 21, 22. Hematopoietic development in zebrafish occurs in 4 distinct waves, the first two are termed 'primitive', generating macrophages and erythrocytes prior to 30 hours post fertilization (hpf). Two 'definitive' waves follow; first, erythromyeloid progenitors form at 30 hpf within the caudal hematopoietic tissue (CHT) or posterior blood island (PBI). At 32–36 hpf the second wave generates HSCs from endothelial cells lining the ventral wall of the dorsal aorta, a region thought to be comparable to the mammalian aorta-gonad-mesonephros (AGM).23, 24 These HSCs subsequently colonize the CHT and the adult hematopoietic organs, the kidney and thymus 25–27.
C-Myb is largely dispensable for primitive hematopoiesis but is essential for definitive blood formation and the normal development of erythroid and megakaryocyte lineages 28. Importantly, c-Myb is also required in non-hematopoietic tissues and c-Myb loss- or gain-of-function assays in vivo result in severe developmental malformation and embryonic lethality which can preclude hematopoietic analysis 29. One approach to circumvent this is by manipulating tissue-specific regulators of c-Myb, which allow analysis of c-Myb's functions within a specific developmental context.
In this study we demonstrate the direct regulation of c-Myb by miR-126 and the importance of the miR-126/c-myb interaction during definitive hematopoiesis in the zebrafish in vivo. We show that precisely regulated levels of c-Myb are responsible for determining the lineage decision between megakaryocyte/thrombocyte and erythrocyte cell fates in the zebrafish. Furthermore, we show that regulation of this cell fate decision depends upon the concerted actions of both miR-126 and miR-150 assigning a permissive role to miR-126 to promote megakaryocyte/thrombocyte lineage commitment in vivo.
Wild-type AB stocks of Danio rerio, the transgenic lines Tg(c-myb:EGFP) 30, Tg(gata1:dsRed) 31, Tg(cd41:EGFP) 32 and the mutant line Nacre 33 were maintained by standard methods 34.
Morpholinos (Gene Tools, LLC, Philomath, OR, USA) were designed to target the mature miR-126 (MO126) and miR-150 (MO150), the Drosha and Dicer sites of pri-miR-126 (MOMB), the putative miR-126 seed sequence in the zf c-myb 3′UTR (c-myb-TPmiR126) and the translational start site of c-myb (MOmyb). The standard control MO from Gene Tools was used as a control (MOctrl). Synthetic miRNA duplexes for miR-126, 150 and 451 were purchased from Dharmacon (Dharmacon Inc, Lafayette, CO, USA) miR-451 was used as a negative control miRNA for target validation studies (miR-ctrl). MOp53 was co-injected with other morpholinos to abrogate non-specific morpholino toxicity 35. Morpholino and miRNA sequences are provided in Table 1. Morpholinos, miRNA duplexes and mRNAs were injected into the yolk of 1-cell stage embryos. MOs were injected at 8ng (MO126, MOctrl), 0.6ng (MO150), 0.8ng(for fluorescence quantification) or 2ng (for ISH analyses) (c-myb-TPmiR126), 3ng(MOMB), 2ng (MOmyb) and 1.6ng (MOp53); MiRNA duplexes were injected at 5-10μM; mRNAs were transcribed from linearized pCS2+ and injected at 350ng/μl (EGFP-c-myb-3′UTRwt and mut), 80ng/μl (mCherry), 100ng/μl (c-myb).
WISH for mRNAs was performed as described previously 36.
Full-length cDNA of zebrafish c-myb and c-myb 3′UTR were isolated from total RNA by one-step RTPCR (Qiagen, Valencia, CA, USA) and cloned into pCRII, verified by sequencing and subsequently subcloned. Primers for full-length zf-c-myb amplification included a 5′ BamHI site and a 3′ EcoRI site to be subcloned into the pCS2+ expression vector. Primers for the c-myb 3′UTR included a 5′ XhoI site and a 3′ XbaI site to be subcloned into a pCS2-EGFP expression vector C-terminal to the EGFP open reading frame. pCS2+. Mutated c-myb 3′UTR was generated using a site-directed mutagenesis kit (Stratagene, Santa Clara, CA, USA). Oligo sequences are provided in Table 2.
Embryos (~150) were collected from Tg(c-myb:EGFP), Tg(cd41:EGFP) Tg(gata1:dsRed) and Tg(gata1:dsRed/cd41:EGFP) at 36 hpf and 4 dpf 31, 32 respectively and disaggregated into a single cell suspension as previously described 26. Sorting of c-myb:EGFP+, cd41:EGFP+ and gata1:dsRed+ cells was performed on a BD FACSAria (Becton Dickenson, Franklin lakes, NJ, USA) and FACS analysis on gata1:dsRed/cd41:EGFP embryo suspensions was collected on a FACScaliber (Becton Dickenson, Franklin lakes, NJ, USA) and analyzed using Flowjo software (Treestar Inc, Ashland, OR, USA).
Total RNA from whole embryos or sorted cells was extracted with TRIzol (Invitrogen, Carlsbad, CA, USA). qRTPCR was performed using miRScipt Reverse Transcription and SYBR Green PCR and miRScript Primer Assays (Qiagen) according to the manufacturers protocol. All qRTPCR reactions were performed in triplicates in 2 independent experiments. Gene expression changes were quantified based on the delta-delta CT method using the relative expression software tool (REST) 37. Oligo sequences are provided in Table 2.
Embryos (50 or more) were lysed in 2μl/embryo lysis buffer (1% Nonidet P40, 0.1% SDS, 100mM NaCl, 50mM Tris pH 7.4, 10mM EDTA, 0.2mM PMSF, complete mini protein inhibitor cocktail (Roche, Indianapolis, IN, USA)), incubated on ice for 5 min. and spun down at 12500g and 4°C for 10 min. Protein concentration of the supernatant was determined using the DC Protein Assay Reagents Package (Bio-Rad, Hercules, CA, USA). For SDS-PAGE, 100μg protein was combined with 4x denaturing Laemmli buffer, heated at 70°C for 10 minutes and loaded on NuPAGE Novex Bis-Tris Mini Gels (Invitrogen) and separated according to the manufacturer's protocol. Membranes were blocked with 5% milk powder in TBST (0.1%) and probed with primary antibody overnight at 4°C (mouse anti-c-Myb- MCA1793; 1:500 (AbD Serotec, Raleigh, NC), rabbit anti-beta-Tubulin; 1:10000- ab6046 (Abcam, Cambridge, MA, USA)). Membranes were then washed in TBST and probed with anti-rabbit or anti-mouse HRP-coupled secondary antibodies; 1:5000 (Perkin-Elmer Life Science, Waltham, MA, USA) for one hour. After washing in TBST proteins were detected by ECL (Pierce, Rockford, IL, USA).
To determine miRNA regulation of the c-myb 3′UTR in live zebrafish embryos we quantified the relative fluorescence upon co-injections of mRNA of the c-myb 3′UTR fused to EGFP, mCherry and various miRs and MOs. Co-injection of mCherry-encoding RNA with the c-myb 3′UTR:GFP mRNA was used in all conditions as an intra-embryo injection control. Relative fluorescence was calculated to account for normal variations observed in expression levels of injected mRNAs between embryos (related to minor variations in injection and translation efficiency). All experiments were performed in triplicate. Five representative embryos from each injection set were selected and imaged on a Zeiss Axio Imager Z1 microscope (Carl Zeiss, USA) using a Hamamatsu Orca-ER (Hamamatsu, Bridgewater, NJ, USA) digital camera and Openlab software (Improvision, Perkin-Elmer). To calculate relative fluorescence for each embryo, identical selections from three different body regions were chosen from individual embryos and measured for the sum of their pixel intensity using Volocity software (Improvision). Relative fluorescence was determined for each embryo (ΣGFP / ΣmCherry). These values were then averaged for all embryos analyzed for a given injection condition, normalized to the average relative fluorescence of the control miRNA-injected embryos, and compared to each other to determine the effect of a given injection condition on fluorescence intensity resulting in a readout for miRNA-mRNA interaction in vivo.
In order to identify novel miRNAs regulating c-Myb, we analyzed the c-myb 3′UTR for potential target sites of miRNAs expressed in HSCs. We were particularly interested in miR-126 because it is highly expressed in normal human and murine HSC and acute myeloid leukemia (AML) leukemia–initiating cells 38–41 and is part of a distinct micro-RNA overexpression signature in core binding factor leukemias 40. Our analysis revealed a putative miR-126 target site within 120 bp downstream of the c-myb stop codon with a perfect seed match that was conserved in three mammalian and two teleost fish species (Fig. 1A,B). Consistent with these findings, the sequence of mature miR-126 was also conserved among these organisms (Fig. 1B), as well as the sequence of miR-150 (Fig. 1C). Notably, mammalian 3′UTRs contained at least two putative miR-150 target sites, while zebrafish and medaka 3′UTRs had a single perfect seed match for miR-150 (Fig. 1A).
We evaluated the function of the putative miR-126 target site using an in vivo reporter assay in zebrafish embryos 22. Figure 2A illustrates our reporter assay where mRNA encoding the wild-type zebrafish c-myb 3′UTR fused to EGFP was co-injected with mCherry mRNA into embryos at the one cell stage. The co-injected mCherry mRNA controlled for injection variability. Changes in levels of EGFP expression relative to mCherry expression were assessed in the presence of various miRNA duplexes (Fig. 2B). Co-injection of miR-126 resulted in a statistically significant decrease (35%) in c-myb 3′UTR-EGFP fluorescence compared to co-injected control miRNAs and indicated potent negative regulatory function (Fig. 2B,C). Expression of miR-150 yielded a similar 30% reduction in EGFP fluorescence. To confirm the specificity of these observations we also performed assays using EGFP fused to a mutated 3′UTR that contained four nucleotide substitutions within the miR-126 target seed sequence, or a target protector morpholino (c-myb-TPmiR126)), which competes with miR-126 for the specific target sequence in the c-myb 3′UTR (Figs. 1D,E and and22)42. In both cases, GFP fluorescence was restored to control levels (Fig. 2B, C). Interestingly, the co-expression of both miR-126 and miR-150 showed no additive effect on the reduction of GFP fluorescence when compared with either of the miRs expressed independently. Target protector morpholinos provide an exquisite means to inhibit specific mRNA/miRNA interactions during zebrafish embryonic development without affecting other potential target mRNAs or miRNAs. Given that the miR-126 and miR-150 target sites in both fish species are in close physical proximity within the 3′UTR, we verified that c-myb-TPmiR126 blocked the miR-126 effects but did not interfere with miR-150-mediated regulation of c-Myb (Fig. 2B, C). Our findings show that miR-126 can interact in vivo with its predicted target site within the c-myb 3′UTR to attenuate translation of an upstream coding region.
To demonstrate the physiological relevance of the miR-126/c-myb interaction, we analyzed c-Myb expression in zebrafish embryos and sorted zebrafish hematopoietic stem/progenitor cells. As in mammals, c-Myb is broadly and dynamically expressed in many tissues, including HSCs of the developing hematopoietic system 20. Definitive c-myb+/runx1+ HSCs appear at approximately 36 hpf within the AGM 43, and 4 hours later begin to migrate into the kidney and thymus, the adult hematopoietic organs, and the posterior blood island (PBI) 25–27. In order to analyze c-myb mRNA levels in hematopoietic stem cells (HSCs) upon knockdown or forced expression of miR-126 in vivo, we used fluorescent-activated cell sorting (FACS) to isolate EGFP+ HSCs from embryos of the transgenic zebrafish line Tg(cmyb:EGFP), which drives EGFP under the control of the zebrafish c-myb promoter 30. MiR-126 knock-down was validated by qRT-PCR using a combination of morpholinos targeting either the mature form of miR-126 (MO126) or a multiblocker morpholino targeting the Drosha and Dicer cleavage sites within the primary miR-126 transcript (MOMB) or both. (Fig. 3B) The combination of both resulted in a robust reduction of mature miR-126 (Fig. 3A,B). A recent report indicated that miR-126 knockdown in zebrafish embryos resulted in a loss of vascular integrity and hemorrhage 44. However, at the morpholino concentrations used in this study, knockdown of miR-126 did not yield any obvious defects in circulation and embryos were viable through 4 dpf with intact circulation (Fig. 4; Suppl. Movies). In contrast, enforced expression of miR-126 by injection of a synthetic miR-126 duplex resulted in gross developmental malformations and circulation failure by 25 hpf, most likely due to the inhibition of c-Myb and other miR-126 target genes during development. Despite their morphological defects, c-myb:EGFP+ cells were formed in these embryos and were FACS sorted to provide data on c-Myb mRNA and protein levels 31 (Fig. 3A). The c-myb:EGFP+ cells from these embryos at 36 hpf revealed an expected inverse correlation of miR-126 to c-myb mRNA levels. However, the observed differences in mRNA levels were not statistically significant (Fig. 3A), suggesting that miR-126 primarily regulates c-Myb protein synthesis, rather than mRNA stability. To test this comprehensively, c-Myb protein levels were analyzed in lysates from whole zebrafish embryos at 24 hpf (Fig. 3C). Enforced expression of the miR-126 duplex reduced c-Myb protein to nearly undetectable levels, while the knockdown of miR-126 by morpholinos effectively increased c-Myb protein levels. A similar increase in c-Myb protein levels was observed when the c-myb-TPmiR126 was used to block the miR-126 target site in the c-myb 3′UTR to disrupt endogenous miR-126/c-myb interactions. We also assayed miR-150 function, which has been shown to regulate c-Myb in human and murine cells 19. The morpholino knockdown of miR-150 resulted in an increase in c-Myb protein while overexpression of miR-150 caused a reduction in c-Myb protein levels, validating a physiological role of the miR-150/c-myb interaction in the zebrafish. Our results indicate that both miR-126 and miR-150 directly regulate c-Myb protein levels in vivo during embryogenesis.
Zebrafish thrombocytes represent the functional equivalent of mammalian platelets 45. Despite this functional similarity, little is known about the ontogeny of thrombocytes in zebrafish. Megakaryocytes, the precursors of mammalian platelets, have not been identified in the zebrafish kidney marrow 20, 46. Nonetheless, many genes essential for megakaryocyte development are present in zebrafish, suggesting that their developmental program is conserved. As a novel regulator of c-Myb, we examined the role of the miR-126/c-myb interaction during thrombocyte-erythrocyte lineage commitment. In situ hybridization assays for mature markers of thrombocytes (tpoR), and erythrocytes (slc4a1) were performed after experimentally altering miR-126 and miR-150 levels (Fig. 4A–L). Functional knockdown of miR-126 by morpholinos targeting the microRNA or inhibition by the target protector morpholino, c-myb-TPmiR126, resulted in an increase in erythrocytes and a decrease in thrombocytes compared to control (Fig. 4A–F). To further demonstrate the dependence of this phenotype on the c-Myb pathway, we concurrently knocked down c-Myb expression by morpholino. While complete loss of c-Myb leads to severe morphological defects 47 titrating its knockdown demonstrated the ability to rescue the erythrocyte-thrombocyte phenotype (Fig. 4G,H). Consistent with studies in mammalian cells 48, forced expression of miR-150 resulted in an opposite effect, reducing c-Myb and increasing thrombocytes while decreasing erythrocyte numbers (Fig. 4I,J). Interestingly, the miR-126 loss-of-function phenotype was dominant over the miR-150 gain-of-function, in that over-expression of miR-150 could not rescue the miR-126 knockdown phenotype (Fig. 4K,L). These findings were confirmed independently by FACS analyses using a double transgenic reporter line Tg(cd41:EGFP):Tg(gata1:dsRed) that expressed EGFP in thrombocytes and dsRed in erythroid cells (Fig. 4M) The ratio of gata1:dsRed+ erythroid cells compared to CD41:EGFP+ thrombocytes was 7:1 in the control injected embryos. This increased to 10:1 when miR-126 was knocked down. Interestingly we see an even further increase in the ratio of gata1:dsRed/CD41:EGFP in the c-myb-TPmiR126 condition. This may reflect more complete inhibition of the interaction between miR126 and its binding site by blocking this directly compared to morpholino knockdown of miR-126 where residual miR may be present. Knockdown of c-Myb combined with knockdown of miR-126 completely abrogated the effects of the miR-126 knockdown, and supporting the in situ experiments, overexpression of miR-150 alone reduced the ratio of gata1:dsRed/CD41:EGFP by half (7:1 to 3.5:1), while it failed to do so in combination with miR-126 knockdown.
Unfortunately, the converse experiments could not be performed because either the overexpression of miR-126 or the knockdown of miR-150 resulted in severe developmental defects that precluded analysis at later developmental stages (Fig. S1 and data not shown). For each of these two miRNAs a number of additional mRNAs contain putative seed sequences. For example, the TargetScan algorithm13 for target prediction yields 18 putative targets for human miR-126 under the most stringent search parameters. For human miR-150 a total of 142 genes are predicted targets containing at least one seed sequence. Thus, the developmental effects of miR-126 overexpression and miR150 knockdown during embryogenesis are likely to arise from altered expression of not only c-Myb but one or more of their other target genes.
Since both miR-126 and miR150 target c-myb and result in altered megakaryocyte-erythroid cell fate, we addressed the relative expression levels of miR-126 and miR-150 in developing zebrafish thrombocytes and erythroid cells using quantitative RT-PCR on RNA extracted from sorted Tg(cd41:EGFP) and Tg(gata:dsRed) from 2dpf to 4dpf. We determined that the level of miR-126 is 6 fold higher at 3dpf and 10 fold higher at 4dpf in cd41-EGFP+ thrombocytes compared to miR-150 while in contrast the levels of both miRs were similar in gata1:dsRed erythroid cells (Fig 4N).
In this study, we have identified miR-126 as a physiological regulator of c-Myb. We show that it is the specific interaction of miR-126 and c-myb at the identified target site that permits post-transcriptional regulation of c-Myb levels. Our observations indicate that the level of c-Myb in hematopoietic tissue is dynamically regulated by the activities of miR-126 and miR-150, and that these effects on c-Myb result in control of the thrombocyte-erythrocyte lineage decision by multipotential progenitor cells. Thus, our data support the conservation of miRNA regulatory mechanisms of c-Myb function during mammalian and zebrafish hematopoiesis. Our data suggest that these microRNAs may act through a sequential rather than an additive mechanism to control the levels of c-Myb and that regulation of c-Myb during the thrombocyte-erythrocyte lineage decision depends preferentially on miR-126 or miR-150 at different developmental stages. Indeed, the level of miR-126 exceeds miR-150 levels at 3dpf six fold and is 10 fold higher at 4dpf in cd41-EGFP+ thrombocytes while in contrast the levels of both miRs were similar in gata1:dsRed erythroid cells. . While our study did not address more specific aspects of vessel formation it is notable that miR-126 has been demonstrated to play a role in vascular development 44, 49. Hemorrhages are a prominent feature of the phenotype observed in miR-126 knockdown in the studies by Fish et al and Nicoli et al. In view of our findings this may be a result of both loss of vascular integrity and thrombocytopenia. In addition, both microRNAs have many predicted targets, including others in the c-Myb pathway. For example, miR-126 has been suggested to target Tom1 (Target of myb 1), which in turn is suggested to activate c-Myb transcriptional activity via modulation of NF-kappaB signaling 50–52. On the other hand, miR-150 is predicted to target Pim1, another positive regulator of c-Myb 53. Hence, in addition to directly regulating c-Myb the complementary but different effects of these two miRNAs may be mediated, at least in part, by other target genes that in turn affect c-Myb function. In summary, this paper adds another level of complexity in the c-myb regulatory pathway, and underscores the importance of animal models where specific subsets of cells can be tracked during development.
We thank R. Hoffmans, U. Pyati and N. Bushati for critical comments on the manuscript and L. Zon for the Tg(c-myb:EGFP) zebrafish line. This work was supported by the NIH grant CA93152 (A.T.L). E.M.P. is the recipient of a Clinical Research Training Fellowship from Leukemia & Lymphoma Research UK. N.B is a Special Fellow of the Leukemia & Lymphoma Society.
Conflicts of Interest
The authors declare no conflicts of interest
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Packet PickupPlease Read Carefully
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Jacuzzi® Hot Tub Mini FX Jet. Part No.6540-555
Jacuzzi® Hot Tub Mini Power Pro FX Jet was introduced in February 2004 and replaced the Mini PowerPro DX jets. Used in the Jacuzzi J300 and J400 Series. Please note this jet does not come with the stainless steel escutcheon. This Jet measures 8.5cm across the jet face.
Part No. 6540-555.


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Palermo San Remo Rustic Kitchen Doors
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MyHospitalityLab with Pearson eText -- Access Card -- for Intro to Hospitality, 6/e and Introduction to Hospitality Management
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Becky McFalls Schwartz
Becky McFalls Schwartz
Beverage DIrector
Becky's first job was in a Japanese restaurant in Utah when she was fourteen years old. Since then, she's gotten a couple of degrees, interned in fields as diverse as sushi preparation, artist biopics, and tattooing, and published essays & food reviews in various New York magazines (ie; Nylon, Brooklyn Magazine). Before coming to Bar Moga, Becky trained under Sasha Petraske at Milk & Honey and Little Branch, where she honed her skills in classic cocktail preparation for nine+ years. In addition to being part of the opening teams of numerous New York bars and restaurants (both behind the bar as well as in management), Becky had the pleasure of working with the special group at Bohemian, and it was through them that she was introduced to Bar Moga. Becky's award-winning cocktails can be found in numerous cocktail publications (ie; Mr. Boston Official Bartender's Guide) and are served in cocktail bars around the globe.


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GSP Wheel Bearing Hub Assembly
- Bears the vehicle weight and provides accurate rotation for power transmission.
- Reduced weight and ease of assembly, combined with ABS system requirements (if applicable).
- Replaces individual components such as bearings, seals and ABS sensors.
- Use of OE harnesses and is "dyno" checked for proper frequency output.
- Tempered for at least 5 hours.
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We offer workshop installation service and self-collection.
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If interested, simply pm me or call me at +65 8200 4155 to find out more!
Carousell: https://carousell.com/automotive.parts/
© 2017 Sgadsonline.com


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SAT NAV, PANORAMIC ROOF, 7 SEATS, 20s
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Behans Bar & the Witton Arms Fighting Cancer
Paul O'Mahony, the gaffer of both the Witton Arms and Behan's Bar is set to take part in a White Collar Boxing event on September 5th at Edgbaston Cricket Ground on behalf of Cancer charities.
The Corkman who will be 52 on 24th August, incidentally also his wedding anniversary with wife Marie, is well into a nine week training regime in preparation for the bout.
Paul is confident that his fundraising efforts will be supported by customers of both of his pubs ad by a wide circle of friends from across the city to further help eradicate the disease.
When asked his reasons and motivation behind the decision to take on the challenge of the fight, Paul said, "A few months ago, a friend and a member of staff, Siobhan was diagnosed with cervical cancer. She is just 21, she has a three-year-old-son but cannot have any more children, because of her treatment, to me this is totally wrong."
Paul continued, 'An old friend, Olive, also from Witton was recently diagnosed with breast cancer and had a breast removed. How does that make women feel? Obviously I cannot answer that.
"Thirdly, on the 1st of January 2007 my brother Denis, who was 55, was diagnosed with cancer. I remember meeting his daughter in the corridor; she had just been told that her father had the disease. I remember standing in that corridor like it was yesterday, hugging and crying together.
When Denis was diagnosed he was a fit, strong man. 51 days later he passed away. It was horrible to watch someone I loved so much fade away and I prayed to God to stop his punishment."
Paul is determined to raise as much money as possible for the charities he is supporting to help the fight against this horrible disease that has affected every one of our lives at some time or another.
Paul added, "Every punch thrown that night is another blow to cancer."
Tickets for this special one off event are available at £20 for the standing area and £30 each for a seat at a ringside table, complete with waitress service for the bar. All tickets are cash on delivery.
Tickets can be obtained from Behan's on Highfield Road, Hall Green or the Witton Arms, Witton Road Aston.
Paul said, "Tickets are selling out fast so don't miss out and Help support Cancer Research."
There will be an after show party at Behan's Bar with live music, a late bar, a raffle and possibly an auction too.
At the time of print Paul is setting up a link on Facebook, Just Giving, and a method to Pay Donations Directly.


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1 Bed, 1 Bath Unit In The Spectacular Harrison Gardens Building
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Willowdale East: A hub for Uptown Toronto with nearby landmarks such as Mel Lastman Square, Yorkdale Shopping Centre, and G. Ross Lord Park. Nearby motorways include highways 401, 407, and DVP. Nearby access to transit includes Sheppard Subway Station and numerous bus stops. A buzzing and vibrant area with a platitude of restaurants, gyms, supermarkets, theaters, library and government buildings.
Additional Details
- Availability: January 1 (Minimum one year lease)
- Location: Toronto (@ Yonge and Sheppard)
- Utilities: Heat and water included. Tenant pays for all other utilities.
- Parking: 1 space included


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Watch as NBA star Kevin Garnett and hip-hop artist Young Jeezy go heads-up in an intense Texas Hold 'Em match. Being held at an adidas Originals party, these two talk trash and try to get in each other's heads during this face-off, leading up to the river by going all in with their pairs of adidas Originals. Find out who really had the nuts in this video.
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St Servanus No. 771, Masonic Lodge, Alva, Scotland
The Freemasons Lodge of St Servanus No. 771 was chartered by the Grand Lodge of Antient Free and Accepted Masons of Scotland on 6 February 1890.
First Published: 28 June 2013 Last Updated: 15 September 2017
Ladies Christmas Nite
Saturday 18th November 2017
Ladies Christmas Night 2017
Click here for further details
Function Suite Hire
Weddings, Engagements, Birthdays, Christenings, Funeral Purveys
Function Suite Hire for your special occasion - seating for 150 persons.
Click here for further details
Lodge Meetings
Masonic Hall, Ochil Rd, Alva, FK12 5J
- - - - - - - - - -
Monday 18 September 2017 - 7:30pm
To be advised
- - - - - - - - - -
Monday 2 October 2017 - 7:30pm
To be advised
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Monday 16 October 2017 - 7:30pm
To be advised
- - - - - - - - - -
Lodge St Servanus No. 771 Freemasons Lodge
We hope that you will find this website informative about Freemasonry in general and Lodge St Servanus in particular.
Firstly a site navigation tip - no matter where you are on our website clicking on the Lodge Crest (top left hand side of every page) will bring you back to this page.
As well as keeping our members, fellow freemasons and non-masonic visitors informed about our history, meetings and social events, one of the aims of this website is to inform the general public about Freemasonry and to increase awareness of who we are and what we do.
Contrary to popular belief, freemasonry is not a secret society. Indeed the opposite is the case as our laws and bye-laws make it compulsory that we advertise the time, date and location of our lodge meetings, usually within the local press - hardly the actions of a secret society!
Freemasonry is a fraternal society that expects a high standard of personal conduct of its members and has a longstanding tradition of care and concern for family, friends and for the wider community.
This link to Frequently Asked Questions will hopefully answer some more questions you may have about Freemasonry.
We hope that the information you find within the pages of our web site will further stimulate your interest in freemasonry and you are cordially invited to make contact with us should you wish to find out a little more.
I extend warmest greetings to all our website visitors.
Bro. Gordon Wm. Guthrie P.M.
Right Worshipful Master


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Montclair State University -Public
Montclair, New Jersey
20,022 (15,885 undergraduates)
Does College Size Matter?
2012, 2013, 2014
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2014-15 Accepted: 67%
4 Year Graduation Rate: 34%
5 Year Graduation Rate: 58%
Why YOU Should Care About College Graduation Rates
Total Expenses In-State: $28,350
Total Expenses Out-of-State: $37,064
Average Net Price 13-14: $14,798
Freshman Non-Federal Loans: 12%
Net Price Calculator
5 Things to Know About Paying for College Before You Apply
75th PERCENTILE TEST SCORES
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SAT Math: 540
ACT: na
The Truth About Scholarships
Percent Freshman not in top quarter of class: 63%
Percent Freshman with GPA <3.50: 77.2%
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Average Freshman Non-Need Award: $4,664
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Passaic River Institute


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Dalian is the second largest city in Liaoning Province, Dongbei (North East), China and the largest port in northern China as well as a major destination for Chinese tourists. Located at the southern tip of the Liaodong Peninsula, the main part of the city is on its own sub-peninsula, with the port to the north of the town centre and natural coastline dotted with beaches to the east and south.
Combined with the city's many parks and green hills, wide thoroughfares and army of street cleaners, make Dalian a more pleasant city to visit and live in than most Chinese cities of comparable size.
There are no articles in this category. If subcategories display on this page, they may contain articles.
Zhongshan Square in the heart of the city's financial district is surrounded by some of the best examples of colonial-era architecture in Dalian. Though the square was first laid out by the Russians most of the buildings around the square were built between 1910 and 1920 by the Japanese. The square itself serves as a social hub, with locals gathering during the evening to play hacky-sack or watch football games on the big screen above the Dalian Hotel. Square is currently under major construction.
Russian Street, Across Shengli Bridge, NE of the train station. One of the few places it's still possible to get a glimpse of Dalian's colonial past. Most of the old Russian and Japanese buildings were razed in the many bursts of redevelopment the city has seen over the years, but the old architecture is "preserved" in the sense many of the buildings along the road are actually modern facsimiles of old Russian buildings rather than the genuine article. The old buildings that do remain are worth examining, however, and more genuine examples lie along the side roads to the east of the main street. Stalls and shops along the road sell various trinkets and souvenirs (no affiliation with Russia other than some wooden dolls) but the same items can generally be found for much less elsewhere in the city. A similar Japanese Street exists in the south of the city center, near the Children's Park, but it is almost wholly fake, with empty shells of buildings lining the road. Those interested in real examples of Japanese houses would be better served exploring the area around Gaoerji Lu to the south of People's Square, although much of what exists is in shambles.
Xinghai Square. On the coast south west of the city center, is the largest square in Asia, a fact frequently mentioned in Dalian tourist literature. There are a number of attractions around the square, including museums, a bathing beach, large aquarium, Xinghai Convention and Exhibition Center, huge indoor tennis court and swimming pool, a number of seafood restaurants boasting local fare, bungee tower and a fairly small amusement park that's very kid-friendly. With the possible exception of the aquarium, museums and restaurants, few of the attractions are likely to be that interesting to visitors but it's still worth taking the time to visit the square and have a wander around the area.


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Product 7/7
393 page manual, 104 page workbook, 6 hour CD-rom, and CD jacket (answer book not shown)
CNC turning centers are among the most popular types of CNC machine tools. Most companies that have any CNC machines have at least one. Unfortunately, companies are finding it more and more difficult to find and hire qualified CNC people. Many are realizing that they must provide extensive training to new hires and provide at least some continuing training to established employees.
This affordable courseware makes it possible to train CNC people from scratch. While the course assumes the student has some basic machining practice experience, but assumes nothing about their previous CNC skills. Using the proven key concepts approach, the course brings students up to speed gradually ? constantly building upon previously presented information ? and stresses the reasons why things are done as importantly as how they?re done. Six of the ten key concepts are most related to programming, and four are related to setup and operation.
We further divide the key concepts into twenty-eight lessons. Lessons range from under five minutes to just over twenty minutes in length (total course presentation time is five hours fifty-two minutes on one CD-rom).
Twenty Eight Lessons:
All examples are shown in the format for the most popular control in the industry ? the Fanuc control. Note that many control manufacturers claim to be Fanuc-compatible.
When the full course is purchased, this courseware includes everything your people need to learn CNC turning center programming setup, and operation - and it can be used over and over again to teach others in your company about CNC turning center usage.
All presentations have been developed using Microsoft PowerPoint which is the industry standard for computer generated presentations. We use the freely distributed PowerPoint Viewer to display all presentations. PowerPoint Viewer is included.
System requirements: You must have a multimedia computer with Windows 98 or above and a minimum of 64 megs of internal memory to run the courseware.
Full Course Includes:


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Designed to give you an intense shine and strong hold, this Oud Royal Gravity-Defying Gel from Philip B is a luxurious gel that's perfect for creating sculpted styles. Working to give you a professional finish, this gel is Paraben, Phthalate and Sulfate free.
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Water (Aqua), VP/VA Copolymer, Glycerin, PEG-40 Hydrogenated Castor Oil, PVP, Fragrance (Parfum), Phenoxyethanol, Acrylates/Beheneth-25 Methacrylate Copolymer, Aminomethyl Propanol, PEG-90M, Ethylhexylglycerin, Buteth-3, Linalool, Hexyl Cinnamal, Sodium Benzotriazolyl Butylphenol Sulfonate, Citronellol, Benzyl Salicylate, Butylphenyl Methylpropional, Bamboo (Bambusa Vulgaris) Extract, Limonene, Amyl Cinnamal, Alpha-Isomethyl Ionone, Hydroxyisohexyl 3-Cyclohexene Carboxaldehyde, Cinnamal, Tributyl Citrate, Yellow 5 (CI 19140),Panthenol, Red 4 (CI 14700), Green 5 (CI 61570), Yellow 6 (CI 15985), Styrax Benzoin Resin Oil, Benzaldehyde, Benzoic Acid, Benzyl Benzoate, Cinnamic Acid, Coniferyl Benzoate, Vanillin.
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Express Delivery Available - If ordered before 9pm, delivered by courier next working day. More Info »
If ordered before 9pm, delivered by courier next working day. Available on orders placed before 9pm (2pm weekends and Bank Holidays) for delivery next business day. Applies to most delivery addresses within mainland UK.
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This article presents multiple role-plays and reframing techniques to help mediators further their training.
Ever wondered why mooting or debating is a very essential part of law student's life? Ever wondered what kind of skills a law student can develop by participating in a moot or a debate?
Mediation Trainer Stuart Watson shares the wisdom of the most important elements of a mediation training gained from training new mediators over the last 15 years.
This blog is an invitation not only to the students taking on the roles of client and counsel at the ICC competition, but also to the judges and mediators, to be aware of the unstated, and often unobserved and unrecognised differences between the parties that, despite the common ground of language and process, may well shape the parties' perception of mediation, commercial relationships, the nature of purpose of mediation, and the design of acceptable outcomes.
Does mediation do any good? Does it make the world a better place?
This is a diagnosis for workplace mediators to include in their trainings.
For us and the colleagues we've worked with for many years, our first premise has always been that self-determination, or what we call empowerment, is the central and supreme value of mediation – a premise probably shared by many in the field.
The Olympics have come and gone with all of the emotion and inspiration they bring. In our recent, fully-subscribed, residential Summer School on mediation skills for leaders, we reflected on the learning from Rio. We watched a video replay of the men's taekwondo -80kg final in which Team GB's Lutalo Muhammad lost to his Ivory Coast opponent in the last second of the bout, giving the latter his country's first ever Olympic gold medal.
You don't get better at listening during conflict by practicing during conflict. You get better at listening during conflict by practicing outside of conflict, where the stakes are lower and it's easier to be on top of your game.
"That's not my problem" are four of the most frustrating words to hear when you're trying to talk through a conflict.
The vast majority of people still have confusions, if not difficulties, in having a clear representation of WHO a mediator is, WHAT is it that a mediator aims to achieve, for WHOM, WHAT he or she does, and maybe more importantly, IF, WHEN and WHY would someone benefit from using mediation and mediators.
Bickering, an argument about trivial matters, is one of those everyday bad habits that feeds the growth of destructive conflict in a relationship.
Our intent in a recent training was to provide new mediators training in parenting plan mediation, we received so much more. Over the course of the weekend, I found myself reexamining some of my beliefs about mediation practice.
This is an instructional video produced by the Stanford School of Business. It covers the process of negotiation by Joel Peterson.
This article describes the benefits I see from taking Collaborative training.
These authors assert that most of the traditional values and styles that guide divorce mediation are still applicable.
In order to be effective, trainee mediators need to unlearn much of what they think they already know.
A lot of people cringe when they even hear the word "conflict", especially at their workplaces. But the truth is, conflict is a natural part of human interactions, and it won't go away even if you close your eyes and wish really hard!
Michael Lang describes an account where he was teaching reflective practice, a practice that enables mediators to be more aware of their own methods and why they are using them.
I love the cookbook that Andrea and 1001 chefs wrote, Cooking Up a Deal: Negotiation Recipes for Success. It's a wonderfully short and fun piece to assign for the last day of class.
When I'm mediating, coaching, or training, there are moments I want to illustrate why resistance builds up. There are moments I want to help someone understand in a quick and visceral way that pushing their agenda relentlessly is contributing to getting things good and stuck.
Reports on what lawyers should know, including the MacCrate Report and Educating Lawyers, regularly list problem-solving, negotiation, and dispute resolution as skills that lawyers should have. Best Practices for Legal Education called for law schools to educate students in problem-solving and in practical wisdom, in order to solve clients' problems effectively and responsibly.
The Health Board South Wales asked us to train their Serious Incident Investigators. Although we know investigating inside and out, this was a new context for us that involved a lot of research and adapting to their needs. And it paid off!
The New York Peace Institute recently conducted a mediation training for the NYPD. They have kindly shared how the training went, tips and tricks for training, and some insights into the cross-over between police and mediation.
The current complexity of our problems, at least in the U.S., is close to the point of overwhelm; for the individuals involved, for the governments that rule them, for the courts that adjudicate them, for the economic systems that support them, and because of the corrupted values that guide them.


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Majestic Abundance is specially designed and created with jade beads and semi precious gemstones. It is also created with a solid full jade bangle, together with polygon 925 sterling silver beads to jazz up its look. Semi precious gemstones and crystals are pieced together with the jade bangle to give a distinctive focal point. Imagine striding down the street to work or over weekend during brunch, it simply jazz up your entire look.
Measurement:Polygon shape 925 sterling silver beads: Approx 1.5cmJade beads: Approx 8mm and 12mmJade bangle: Approx 7.4cm acrossCentre piece - Semi precious gemstoneswith crystals: Approx 5cm
Length of entire necklace: Approx 34inch
Colour of jade: Mixture of dark and light greenMaterials:Made with mixture of 925 sterling silver and stainless steel.Material used on jade pieces & semi precious gemstones: 925 sterling silverChain: Stainless steel
One of the Designer's Favourite Pick
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Let everyone know you're ready for the apocalypse with Zombie Outbreak Response Wall Clings. Just peel n' place these Zombie Wall Clings to any smooth, clean surface.
Includes 2 clings
Round cling measures 11" in diameter
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Please allow ample time for delivery. Our estimated delivery date for this product is noted in checkout. Many items are shipped separately, so please allow extra time for these items to arrive. Shipping charges are based on the value of the merchandise and not the number of shipments. For additional shipping information, please contact our Customer Service Department at (email@example.com or 1-800-314-8794).


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Chocolate Caramel Bars Recipe
- 1 cup all-purpose flour
- ½ cup + ¼ cup brown sugar
- ¼ cup + ⅓ cup salted butter
- ½ cup semi-sweet chocolate chips
- Heat oven to 350 degrees F. Spray an 8x8 glass pan with non-stick spray.
- In a medium bowl, combine flour, ½ cup brown sugar and ¼ cup butter. Use 2 forks or a pastry cutter to combine, until mixture is crumbly. Press into bottom of 8x8 pan.
- In a small saucepan over medium heat, melt remaining ⅓ cup butter. Add in ¼ cup brown sugar and stir. Bring to a boil, then boil for 1 minute. Remove from heat and pour over crust.
- Bake for 15 minutes. (Do not over bake or you will have toffee bars!
- Remove from oven. Sprinkle chocolate chips on top of caramel and swirl gently with a knife. Let cool completely. Cut into bars and ENJOY!!
Source: Chocolate Caramel Bars
Chocolate Caramel Bars Recipe Reviewed by kuji hara on 2:10 AM Rating:


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Posts Tagged 'ROI'
What if You Could Combine Outbound/Offline Marketing with Inbound/Online Marketing?
By John Joyce on October 28, 2010 - Comments 2
If you have a local business and still rely on direct mail to reach prospects as well as existing customers, then you have experienced the fear of not knowing whether you have achieved any ROI. What if you could actually track the audience response to your direct mail campaigns and even follow their progress through your own online portal? What if you could track their referral activity once they transitioned into the online world and allow them to utilize social networking links to evangelize your offer to their network of contacts? And, finally, what if you could reward these customers based on their influence and ability to drive new customers to your business? That would be just Dukky!
I read an article about a company called Dukky in the July 2010 issue of Entrepreneur Magazine where the founder of Dukky, Shawn Burst, had an idea to revive the dying direct marketing and print industries. Let's face it, more than 99% of "junk mail" goes directly into the trash. But, with this solution, you have a lead generation system that marries the offline marketing world with the online marketing world. Take a look at the video below to get a better understanding of exactly how Dukky works:
"According to the Direct Marketing Association, more than 54 percent of all advertising spending in the United States goes into direct-marketing channels. Spending in 2009 was more than $149 billion; direct mail and catalogs alone made up $44.4 billion of that."
Businesses spent more than $44B in 2009 on direct mail and catalogs and they have no idea what kind of return they're getting on their investment. Now, Dukky will allow them to develop direct mail campaigns that can be tested and tracked for effectiveness.
For the small businesses that use postcards to remain top-of-mind with prospects, now you can entice them to participate in a more interactive process, to better understand your USP and to share a special offer with their social network. Dukky's small business solution is currently in beta and pricing starts at $99/month so I suggest you get in touch with them and find out how they can help you leverage their solution to grow your business.
The reports of direct marketing's death have been greatly exaggerated – according to Shawn Burst at least.
Why Aren't You Measuring The ROI Of Your Online Marketing Efforts?
By John Joyce on May 28, 2010 - Comments 0
I recently participated in Omniture's "2010 Online Analytics Benchmark Survey" and received a customized report from them yesterday and wanted to share some of the information with you. It doesn't take a genius to understand that businesses shouldn't spend money on new marketing channels without having the ability to track their success, but that's exactly what the majority of us are doing.
The main takeaways from this survey are as follows:
Key Web Analytics Challenges Include Talent, Measuring ROI and Multi-Channel Support
Top challenges for online marketers include:
- Difficulty finding and training top talent
- Determining what actions to take based on their data
- Maximizing marketing ROI through full funnel measurement and automatic multi-channel integration.
Do you find yourself or your staff spending large chunks of time developing programs and campaigns, delivering them across an assortment of online marketing platforms and then wondering if they had any impact? It looks like you're not alone. The most telling statistic from this study is glaring proof that, since many of the emerging online marketing platforms are very much in their infancy, businesses are unable to track their success as illustrated below:
Marketers know which metrics are important to measure, however they are not capturing all the metrics they need to achieve their goals. Eighty-two percent of survey responders believe ROI is somewhat to very important to measure, but only 30 percent of marketers can effectively measure marketing ROI! Additionally, 86 percent of respondents think conversion rate is somewhat or very important to measure yet 27 percent of marketers cannot effectively measure conversion rates.
We all know measuring the success of offline marketing is difficult to nearly impossible but that's because there's no system available to allow for granular collection of metrics. Conversely, online marketing offers so much data that we become overwhelmed by a never ending stream of seemingly valuable information . They key, however, is knowing how to interpret that information and turn it into an actionable plan that exploits the repeatable successes you have unearthed.
If you're going to "try" a mobile marketing campaign, social media campaign or viral video campaign, make sure you develop a system to track and measure success against your business goals. Work smarter and not harder.
Have a great Memorial Day weekend!
Building Your Online Marketing Ecosystem Part VII: Feedback And Comments
By John Joyce on March 16, 2010 - Comments 0
If you want to develop the "persona" of your small business, you have to make sure your audience is easily able to interact and share their thoughts and feedback with you. It's also important to deliver engaging and, in some cases, polarizing and provocative content that will motivate people to interact. Finally, once you spark conversation and debate, you still have to maintain some semblance of control over the process so I have listed several tools available to help with this process:
Introducing The Interactive Online Marketing Ecosystem
By John Joyce on March 11, 2010 - Comments 1
In an effort to offer the most value to our readers, we have created an interactive map of the Online Marketing Ecosystem. There are so many marketing technologies available that we felt it was important to present your options in one central location. This will be a "living" map and will be updated as often as needed – as long as you're willing to share your ideas and suggestions with us.
Here are some simple navigation tips:
- Click and drag to move around the map
- Hover over the small arrow next to the name of each tool to see/follow the external link
- Click zoom in/out buttons at bottom of the window
- Click "full screen" button to view in a browser window
Need Real Time Analytics? Announcing Small Biz Insight
By John Joyce on February 25, 2010 - Comments 0
Small Biz Insight is a real time web analytics service allowing you to view up to the minute data on the traffic to your web site. Most services don't allow you to make split second decisions because of a substantial delay in their delivery of your website data.
Real time data lets you react to changes in your traffic as they occur. For example, if you had an article that hit the front page of a popular site like digg.com, you would see the traffic spike in Small Biz Insight immediately, along with links back to the sources sending you the traffic. Knowing this, you could make changes to your site or to the article itself to take advantage of the situation.
If instead you relied solely on a service like Google Analytics, it would be up to 24 hours before you even knew about the traffic, and by then it would be too late to do anything about it.
Here is a partial list of the real time information available to you in Small Biz Insight:
- Customizable dashboard
- Filtering and segmentation
- Twitter analytics
- iPhone version
- Goals and conversions
- Campaign tracking
- Support for Google Analytics tags
- Information on each individual visitor
There are two versions of Insight available – Self Serve for $9.95/month and Managed for $19.95 which will include customization and reporting services. If you're interested in learning more, please send us an email at firstname.lastname@example.com or call us at (800)277-1187.
If 200K People Retweet Your Promotion And Nobody Becomes A Customer, Is That A Success?
By admin on July 17, 2009 - Comments 4
Too many times we see businesses haphazardly throw random campaigns at the wall to see what sticks. One member of the marketing team is "trying out" a few Facebook ads while a colleague is independently "testing" a viral program on Twitter. I continue seeing articles written about "Twitter done right" and "viral marketing success stories" and not once has there been any mention of ROI or CPA. There are too many marketers patting themselves on the back for simply creating buzz. When did it become acceptable to spend money to get people talking about your giveaway but not your unique selling proposition?
Lisa Barone shared a story earlier this week about a contest giveaway run by a company called Moonfruit celebrating their 10th anniversary. The numbers were quite high – more than 200,000 posts per week. Adam Ostrow over at Mashable wrote, "Not surprisingly, this promotion is working. #moonfruit is Twitter's top trending topic today, beating out the likes of Michael Jackson, #iranelection, and Wimbledon-related tweets."
This promotion is working? How are these people measuring success? Can you imagine approaching your boss and saying, "I'd like to spend $10K+ and use countless company resources so we can be 'bigger than Michael Jackson' for a week" on Twitter. When asked for your ROI estimate you smile wryly and say, "Zero, BUT, hundreds of thousands of people who will never become customers will be talking about the Macbooks we're giving away".
This is the equivalent of a restaurant spending $10k for a big neon sign that says, "restrooms open to the public".


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Jessica Jaymes shows off the charm and sex appeal that have made her a porn industry mainstay. The busty brunette faces the lens and strips without modesty. Her top falls away from her big breasts to show that she has tan lines but no bra. Her nipples are rock hard and aching to be played with. Jessica then loses her skirt and her panties quickly follow. She shows her bald pussy and pulls to spread the lips. She shows her sexy piercing without shame.
Watch Full Scene View the Gallery


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In our year of operation we have featured some very new-school music and we have had djs and bands that took it back to the old school. Our sense of community is reflected in the breadth of underground musical talent that passes through our doors and throws down on the floor at the Works. No sound is taboo and no genre is left unturned. It is also displayed in how we connect with several of the Bay Area's truly energizing and artistically productive non-profits. Sharing with the community is one the things that defines Public Works'…well…public works.
This month Public Works has been given the opportunity to take it back to school period — no reference to old or new — because September is Back to School Month!!! A portion of our takings for the entire month will be given to local organizations, Mission Graduates and Seven Teepees Youth Program. These Mission based groups are going all out to help today's youngsters in a truly positive way. They strive to give the city's youth the support and direction they need to realize their dreams and aspirations.
Mission Graduates goes beyond orthodox methods and realizes a bond between student, family and community. They are, in their own words, "building a community where every young person has the opportunity to discover their personal pathway through education." While Seven Teepees Youth Program is devoted to "impacting the lives of promising inner-city youth." Their mission is to work with the urban youth entrusted to their care in a way that will foster the skills they need to make lifelong positive choices and to create their own opportunities for success.
We feel that these are noble aims, especially in a time when the challenges for kids in urban areas grow ever greater. So if you're in the city, you're a regular at our spot and you want to see the young team get all the help they need (remember all the help you got as a young 'un), when you join us in September your small dance steps will do something giant for the kids. So come on through and get on the floor at the Works.
Chris Orr


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Aftermarket 23 inch wheels for sale will enchance your car performance
Most of the vehicles are equiped originally with 16 inch and 18 inch wheels, but the owners of trucks often look for aftermarket 23 inch wheels for sale. These alloy wheels provide the better performance of your car, they look stunning, and add a luxury look to your vehicle. Allure Custom Automotive offers 23 inch wheels and tires of popular brands.
Ace Alloy Wheels, Foose, Niche Wheels and other manufacturers offer their wheels at this store. The price starts from $363. The shipping of the chosen wheels or 23 inch rims and tires package will be free. You have only to pick up your wheels' brand or tires you want to see on your rims. Toyo, Good Year, General, Hankook tires cost less. Ordering at the site 23 inch rims and tires, you may save up to 50%. Some drivers prefer marketplaces, shopping at Amazon and Ebay. If you have got a positive experience of online shopping already, you will do it again, and get cool cheaper wheels for your vehicle at online stores. You can make a short "voyage" around the local stores checking and comparing the prices given at retailers with online propositions.
OEM rims are original, but who does not want to enchance the look and performance of his wehicle with cool aftermarket wheels? Forged Porsche Cayenne 23 inch wheels, ordered at Vivid Racing or Ebay will make your car looking absolutely stunning!


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4 Stars Hotel
Chablis Palenque
The hotel offers an outdoor pool and rooms with free Wi-Fi. The hotel guests can use the shuttle service available to explore the surrounding area.
Lowest price guarantee
4 Stars Hotel
Ciudad Real Palenque
With stunning architecture and a service level of the best resorts in Mexico, becoming perfect retreat in the Mayan World. Located just 10 minutes from the archaeological zone, 3 and 5 from airport to.
2 Stars Hotel
Posada Aguila Real
Simple rooms with private bathrooms. The Palenque Archaeological site is a 10-minute drive away. Each air-conditioned room, TV, free Wi-Fi is available in public areas. Palenque Bus Station is 4 block.
Lowest price guarantee
If you wish to have your business here contact us.


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ArtPrize Spotlight 2012 – Early Installation Gallery [Photos]
It's only a few days until ArtPrize officially opens all across Grand Rapids, but you can already see the city transforming into a massive art gallery. Installations are cropping up on the Urban Bridge, in The B.O.B. parking lot, near the Gerald R. Ford Museum, and countless other places. In the gallery you will find pieces in all the venues mentioned.
ArtPrize 2012 Spotlight – A Look Back At The Top 10 Entries From ArtPrize 2011
ArtPrize makes September an exciting time in Grand Rapids. Art installations pop up all over the city throughout the entire month. All of that excitement culminates to the top 10 showdown in early October. The top 10 entries can be anything from massive sculptures, to 3D art, to paintings, to pencil drawings. With ArtPrize kicking off on September 19th, let' take a look back at the winners from last year.
Kids Eat Free in Grand Rapids – Best Restaurant Deals
When you are looking for a place to have dinner, the cost is usually taken in to consideration. We realize that, and want to make your decision making process a little bit easier. That's why we put together a list of where kids eat free in Grand Rapids.
West Michigan Remembers 9/11 [Videos]
It has been 11 years since 9/11 and West Michigan honors all those that lost their lives in the tragic events.
New York Prepares For The 11th Anniversary Memorial Of 9/11 [Gallery]
New York pauses each year on the 10th of September to get ready to honor the brave men and women that died in the September 11th attacks. Below you will find a gallery of images of residents of New York taking that pause.
The 7 Best Cat Talking Videos On YouTube
"Meow," it's more than just a noise. It's a way to communicate for cats. But, sometimes their meows can sound a little bit like actual words. In this...
Henri Wins The Internet Cat Video Festival
In other news... There is an Internet Cat Video Festival. The festival was held on August 30th at the Walker Open Field in Minneapolis, MN. Thousands of internet cat videos were submitted under a variety of CATegories like comedy, documentary, animated, etc. The category that held the biggest amount of clout was the "People's Choice" award which was won by Henri, the existential cat.
Polling Places And Voter Information For Election Day 2012 In Grand Rapids
The national election is on November 6th, 2012. We want to help you get all the information you need for the election. You will find the qualifications for voters, a voter registration form, and all of the polling locations for Grand Rapids.
The Chihuly Garden And Glass In Photos – Seattle, Washington [Gallery]
Recently our resident amateur photographer, Gene Parker, was in Seattle, Washington at Chihuly Garden and Galss. The famed artist brought a large portion of his work to the Fredrick Meijer Gardens a few years ago. The sheer amount of glass on display in Seattle is astonishing. Sit back and enjoy a full series of photos that highlight Chihuly's genius.
Channel 95-7′s World's Largest Dog Wash [Video]
Thanks again to everyone who came out to 5/3rd Ballpark on Sunday, August 12th for the World's Largest Dog Wash to benefit Gilda's Club Grand Rapids! We washed 259 dogs and raised a ton of money for Gilda's Club in the process. Check out an awesome video of our furry friends after the jump!
The Top 5 Most Popular Fictional Sharks of Modern Times[Videos]
It's time to celebrate Shark Week and we wanted to give you something a little out of the ordinary. So, behold... The Top 5 Most Popular Fiction Sharks of Modern Times!...
Gunman Shoots "Multiple" People At Texas A&M University
Police have apprehended a man they think shot "multiple" people at Texas A&M University in College Station, Texas. CNN is reporting numerous law enforcement officers were among the victims. The College Station Police department said they don't have a definitive number of victims at this time. The shooting took place about a block from campus, and at least one civilian was shot in the attack.


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124. Rose Marie BRAATZ was born on Jun 25 1922.
She was married to Kenneth FEHRMAN on Jul 18 1940.
Kenneth FEHRMAN died in Feb 1987. Rose Marie BRAATZ and Kenneth
FEHRMAN had the following children:
+175 i. Kathleen Marie FEHRMAN.
+176 ii. William Kenneth FEHRMAN.
+177 iii. Karen Ann FEHRMAN.
+178 iv. Kay Dawn FEHRMAN.
She was married to Herbert ZINCK on Aug 12 1988. Herbert ZINCK was born in Oct 1921.
Return to Table of Contents


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Mountains are particularly special. There's a certain magic to them. Perhaps it's because they can be dangerous or maybe it's because they have the ability to make us feel small. Mountains are teachers. They will instil the virtues of patience, persistence, and gratitude.
Adventure is abound among African peaks. Whether you're ascending one of the many gruelling African peaks or comfortably watching the sunrise over majestic peaks, raw serendipity surrounds you.
No African peaks are the same. Each presenting unique adventures, challenges and varying moments of clarity. From the strenuous, yet rewarding, challenge of conquering Africa's highest Mountain, Mount Kilimanjaro, to soaking up a sunrise from behind the towering Mount Kenya. From the rolling, jagged peaks of the Drakensberg mountain range to South Africa's own natural wonder of the world – Table Mountain, African peaks offer an array of adventure and exploration opportunities that our adventurists keep discovering.


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Words that end in fccr
Found 3 words that end in fccr. Browse our Scrabble Word Finder, Words With Friends cheat dictionary, and WordHub word solver to find words that end with fccr. Or use our Unscramble word solver to find your best possible play! Related: Words that start with fccr, Words containing fccr
- Scrabble
- Words With Friends
- WordHub
- Crossword


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If you HATE your job or your boss so much, why do you work there? Why not look for another job that doesn't make you so miserable?
Asked by Anonymous at 2:48 PM on Mar. 30, 2011 in Money & Work
Answer by Nitestalker at 10:21 PM on Apr. 1, 2011
Answer by rkoloms at 6:27 PM on Mar. 30, 2011
Answer by But_Mommie at 2:50 PM on Mar. 30, 2011
Answer by celeb151 at 2:51 PM on Mar. 30, 2011
Answer by tasches at 6:28 PM on Mar. 30, 2011
Next question in Money & Work
I need to find about $1000 FAST any suggestions?
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Vampire warsFeb. 5th, 2009 06:55 pm
What is with people who take silly online roleplaying games way too seriously? Since I can't play The Sims 2, I've been wasting time playing a Facebook game called Vampire Wars. It's your basic loop of kill, gain abilities, gain skill points, repeat.
Well, I dusted some whelp who got pretty bent out of shape over it - sent me a message telling me not to fuck with her and everything. Judging by the tiny Facebook profile photo, it looked like I was dealing with a teenage girl. This child was apparently under the impression that a game called Vampire Wars would be all fluffy clouds and cuddly kittens and how terrible that her character got killed - how could that ever happen? So, I did what any self-respecting jagoff would do in the situation: I bided my time to wait for her character to regenerate and then killed her off again...repeatedly.
I know I'm an asshole, but in many ways I've ceased to care.


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Industrial-Quality Steel Wool Hand Pad, #4 Extra Coarse, 16/Pack, 192/CartonItem#: YYAZ-GMA117007 | Model#: GMT 117007
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Manufactured by: Seventh Generation
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- Seventh Generation logos are registered trademarks of Seventh Generation. All others trademarks and copyrights mentioned herein are the property of their respective owners.


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Texas Department of Criminal Justice
The TDCJ Commissary and Trust Fund Department utilizes an online program where friends and family may make purchases for eligible offenders up to the maximum quarterly spend. Offender spends are based on the following months:
Up to 100 of the top selling items are available for purchase including snacks, hygiene products, correspondence supplies as well as some new packaged items. The online amount purchased will not affect the amount that offenders may spend on commissary items from funds deposited in their inmate trust fund account. In addition to offenders on commissary restriction, offenders in General Population and State Jail levels 4 and 5, Administrative Segregation levels 2, 3 and transition, Death Row levels 2 and 3 and certain medical conditions are not eligible to receive commissary purchases through this program. (See question 5 under Frequently Asked Questions for full eligibility requirements.)
Friends and family can choose to make an online merchandise purchase or an online deposit to an offenders inmate trust fund account or both. The online merchandise purchase will be delivered to the offender within five business days after TDCJ receives the purchase transaction details from Texas.gov. Please note that unit security or staffing issues can override this timeframe. The on-line deposit to an offenders inmate trust fund account will be transferred to the offenders inmate trust fund account on the second business day from the day of deposit. On-line purchases are subject to substitution, if the item is unavailable at the time of distribution.
The online purchase program is available for all eligible offenders. Commissary staff does NOT determine or control eligibility. An offender may submit an I-60 for eCommDirect eligibility status and may choose to disclose this information to you. Friends and family may contact the TDCJ eCommDirect department at 936-437-4726 or email email@example.com with any questions.
1. How do I place an eCommDirect order?
2. Will I receive confirmation that the items purchased were delivered?
3. How can I tell if my package has been delivered to the offender?
4. May a newly received offender receive purchases?
5. What criteria are used to determine eligibility to receive purchases?
NOTE: Only one purchase per offender may be completed per day. (If a friend or family member has locked the offender out of the system by not exiting the store website correctly, the account will reset on the next business day.)
6. When I am shopping and hit the "Back" button, why am I locked out of the system?
7. Will the eCommDirect purchase amount affect what the offender may spend in the commissary?
8. Is the limit $60 per purchase or $60 per quarter?
Multiple purchases may be made for the offender throughout the quarter but there will be a $3.75 fee assessed for each transaction.
9. May I place an order over the phone?
10. Will I be charged a fee for making a purchase or deposit?
11. Why does the system time me out?
12. Do I have to make a purchase and a deposit at the same time?
13. If I have questions while making a deposit or purchase, who can I contact?
14. Can I make a purchase using any online package program?


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About me
As a Cultural Care local childcare consultant (LCC), I invite our host families and au pairs as well as those considering Cultural Care Au Pair to explore my site and learn about what is going on in our community. I provide year-round support to host families and au pairs, and also talk to families considering the program to help them determine if Cultural Care Au Pair is the right childcare solution for them.
Childcare You Can Trust Like Family!
I love my job as Top National Au Pair Consultant and Matching Specialist – matching families with live-in au pairs they can trust! I enjoy promoting and bridging the gap with childcare for busy, modern families nationwide – with this Global Cultural Exchange Program. I'm honored to be a liaison for the wonderful host families and amazing au pairs in my area; I provide local support, guidance, and coaching to ensure it's a rich and rewarding experience for all – and I'm honored to have received excellent positivity ratings on all Host Family and Au Pair Satisfaction Surveys.
So far, I've welcomed over 100 new host families onto the program – nationwide – as well as helped them find the right au pair for their family! I was one of the distinguished top ten marketers for 2015 (out of over 800 consultants) in the nation and achieved the prestigious Golden Heart Leadership Level and earned 2 International Adventures (Costa Rica and Greece). It was a privilege to be showcased in a 2015 training video at our National Training Conference in Boston – and an honor to be chosen as a mentor for "Make the Leadership League 2017." For 2016, I was privileged to be a speaker on Collaboration & Teamwork at our National Training Conference in Denver. In 2016, I became one of top 1% marketers in the nation and achieved the illustrious Platinum Heart Leadership Level and earned 2 International Adventures (Dominican Republic and Prague).
I live in the Bay Area with my husband and our three awesome children! We enjoy sharing our love for international travel with our children! In 2013, we traveled to Italy; in 2016, we traveled to Mexico; and in the spring of 2017, we're heading to Prague! As an Au Pair Consultant for Cultural Care, I love meeting and welcoming our wonderful, experienced, and talented group of au pairs to the area – and my kids love meeting them, as well!
Please contact me directly (email@example.com ~ 925-719-9767) for my personalized, expert support – I'm here to help you learn more about the rich and rewarding benefits of welcoming an au pair into your home and your heart; as well as help you find the right au pair for your family! Sign up here for my Host Family & Au Pair News and stay connected!
connect with me on
facebook | instagram | linkedin | pinterest | twitter
Let's Chat!