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Hyashi, M., Deng, L., Chen, M., Gan, X., Shinozaki, K., Shoji, I., and Hotta, H.
Notes: The effect of SMYD3 and ERK2 on hepatitis B RNA expression was examine in tranfected Huh-7.5 human hepatoma cells. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System followed by dye-based qPCR analysis. The effect of SMYD3 and ERK2 on AP1-dependent and NF-κB-dependent gene expression in the cells was examined with the pGL4.44 [luc2P/AP1 RE/Hygro] and pGL4.32 [luc2P/NF-κB-RE/Hygro] Reporter vectors using the Dual-Luciferase® Reporter Assay System with a pRL-TK Renilla luciferase control. Reporter assays were read with a GloMax® Instrument. (4603)
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GoScript™ Reverse Transcription System
Dual-Luciferase® Reporter Assay System
pGL4.44[luc2P/AP1 RE/Hygro] Vector
pRL-TK Vector
pGL4.32[luc2P/NF-κB-RE/Hygro] Vector
ReliaPrep™ RNA Cell Miniprep System
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Ratnoglik, S.L., Jiang, D.-P., Aoki, C., Sudarmono, P., Shoji, I., Deng, L. and Hotta, H.
Notes: Primary splenocytes were isolated from mice immunized with an NS3 expression vector and examined for IFN-γ expression by RT-qPCR. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and converted to cDNA with the GoScript™ Reverse Transcription System. The affect of wildtype and mutant NS3 on interferon-β promoter activity with an IFN-β promoter firefly luciferase vector and pRL-TK vector control was measured with the Dual-Luciferase® Reporter Assay System. LDH release was measured with the CytoTox 96® Cytotoxicity Assay in a cytotoxic T-lymphocyte assay using cultured splenocytes from the immunized mice. (4609)
CytoTox 96® Non-Radioactive Cytotoxicity Assay
Guo, R., Abdelmohsen, K., Morin, P.J., and Gorospe, M.
Notes: The let-7 microRNA family are thought to act as tumor suppressors. Let-7 activity is downregulated in several cancers, and overexpression of let-7 inhibits cancer growth in some mouse models. The authors of this paper describe a sensitive luciferase-based reporter assay for detecting let-7 miRNA activity in cells. The reporter construct was based on the pmirGLO Vector, which contains firefly luciferase as the reporter gene and Renilla luciferase as an internal control. The authors inserted let-7 miRNA target sites at the 3′ end of the firefly luciferase gene. Interaction of let-7 miRNA with these target sequences resulted in reduced luciferase activity. The authors used the reporter construct to screen a kinase inhibitor library for compounds that repress let-7 activity in ovarian cancer cells, and identified GSK-3β as a potential target for therapeutics. (4406)
pmirGLO Dual-Luciferase miRNA Target Expression Vector
Yao, T., and Lin, Z.
Notes: This study identified that miR-21 is significantly overexpressed in human cervical squamous cancer tissues and cell lines, and showed that the level of miR-21 correlates with tumor differentiation, and regulates proliferation, apoptosis, and migration of HPV16-positive cells. The authors used gene expression profiling and luciferase reporter assay to identify candidate target genes for miR-21. Wildtype and mutant 3′-UTR sequences from the target gene CCL20 containing putative binding sites for miR-21 were subcloned into the psiCHECK-2 Vector and used to transfect HEK293 cells. The Dual-Luciferase® Reporter Assay System and GloMax® Multi Luminometer were used to measure luciferase activity from both constructs. (4192)
psiCHECK™-2 Vector
GloMax®-Multi Luminescence Module
Cock-Rada, A.M., Medjkane, S., Janski, N., Yousfi, N., Perichon, M., Chaussepied, M., Chluba, J., Langsley, G., and Weitzman, J.B.
Notes: These authors investigated the role of matrix metalloproteinase (MMP-9) in a reversible model of cancer that is initiated by infection with intracellular Theileria parasites. They found that gene induction by parasite infection was associated with trimethylation of histone H3K4 (H3K4me3) at the MMP-9 promoter. The H3K4 methyltransferase SMYD3 was the only histone methyltransferase upregulated upon infection. They therefore investigated the role of SMYD3 overexpression on MMP-9 expression and cell migration, identifying SMYD3 as an important new regulator of MMP-9 transcription. During the study they used the GloMax® Multi Luminometer to measure luminescence and absorbance in reporter and cell viability assays. They also used the Dual-Luciferase® Reporter Assay to measure SMYD3 activity in cells transfected with a SMYD3 reporter, and the pGL4-hRluc/TK plasmid for normalization of the experimental reporter activity. GoTaq® DNA polymerase was used in semi-quantitative RT-PCR. (4189)
Dual-Luciferase® Reporter Assay System 10-Pack
pGL4.74[hRluc/TK] Vector
GoTaq® DNA Polymerase
Yan, C., Zhu, M., Staiger, J., Johnson, P.F. and Gao, H.
Notes: RAW 264.7 mouse macrophage cells were plated in 12-well plates at 1.5 × 105 cells/well for transient transfection with 0.5μg of DNA and 1.5μl of FuGENE® 6 Transfection Reagent in 50µl of Opti-MEM I medium. After 24 hours, the cells were treated with IgG IC for 5 hours and luciferase activity analyzed using the Dual-Luciferase® Reporter Assay System. (4402)
FuGENE® 6 Transfection Reagent
Dual-Luciferase® Reporter 1000 Assay System
Ning, B., Liu., G., Liu, Y., Su, X., Anderson, G.J., Zheng, X., Chang, Y., Guo, M., Liu, Y., Zhao, Y. and Nie, G.
Notes: The authors used GoTaq® DNA Polymerase to amplify cDNA generated from total RNA (RT-PCR) extracted from murine erythroid leukemia (MEL) cells and mouse erythroid burst-forming units (BFU-Es). These cells were used to study the molecular mechanism of 5-aza-2'-deoxycytidine (5-aza-CdR)-induced erythroid differentiation, a process involved in azanucleotides for treating myelodysplastic syndromes (MDS) that reduces the risk of transformation to acute myeloid leukemia (AML). Treatment of these cells with 5-aza-CdR, a hypomethylation reagent, upregulated genes responsible for heme production and iron uptake. The pGL3 basic vector and promoter were used to create plasmid constructs of different E-box regulatory sequences with a luciferase reporter. The plasmids were cotransfected with c-Myc, Max or both transcription factors into human hepatocytes (HepG2). The Dual-Luciferase® Reporter Assay System was used to identify that the –6kb E-box of the transferrin receptor 1 (TfR1) promoter was a strong enhancer for inducing TfR1 expression when c-Myc and Max formed functional complexes that bound to it. Bisulfite sequencing was performed to study methylation patterns after 5-aza-2'-CdR treatment using the pGEM-T® Easy Vector system to ligate the isolated DNA fragments for TfR1 and Fech (ferrochetalase), which were transformed into E coli. for final sequencing. (4176)
pGEM®-T Easy Vector System I
pGEM®-T Easy Vector System II
pGL3-Basic Vector
pGL3-Promoter Vector
Soldà, G., Robusto, M., Primignani, P., Castorina, P., Benzoni, E., Cesarani, A., Ambrosetti, U., Asselta, R. and Duga, S.
Notes: To confirm the role of a mutation in the miR-96 microRNA (miRNA) associated with an autosomal dominant hearing lost, HeLa cells (250,000 cells per well in six-well plates) were transfected with 4µg of plasmid carrying wild type or mutant miR-96 miRNA using FuGENE® HD Transfection Reagent. After 24 hours, the cells were washed and total RNA extracted. After quantitation, the RNA used in RT-PCR analysis. The entire 3´UTRs of eight putative target genes were amplified by PCR from genomic DNA and cloned into the psiCHECK™-2 Vector. HeLa cells were transiently transfected with 2µg of the 3´ UTR psiCHECK™-2 constructs and 0.2µg of a wild-type, single or double mutant miR-96 plasmid using FuGENE® HD Transfection Reagent. Forty-eight hours after transfection, the Dual-Luciferase® Reporter Assay System was used to quantify the firefly and Renilla luciferase in cell lysates. (4251)
FuGENE® HD Transfection Reagent
Xie, Y., Rosser, J.M., Thompson, T.L., Boeke, J.D., and Wenfeng, A.
Notes: This paper describes a rapid dual-luciferase-based assay for L1 retrotransposition that is amenable to high-throughput screening. A firefly luciferase vector in which the luciferase gene was disrupted by an antisense intron was constructed by introducing a 900-bp fragment of the human γ-globin intron into pGL4.13. This Fluc gene, interrupted by an antisense intron, gives only minimal luciferase expression unless the luciferase gene is restored by a retrotransposition event. The authors also tested a similar retrotransposition reporter using the pGL4.73 Renilla luciferase vector, but found that the firefly construct gave much higher signals. They therefore used the firefly luciferase retrotransposition reporter, a Renilla luciferase normalization control and the Dual-Luciferase® Assay to characterize profiles of retrotransposition by various human and mouse L1 elements, and to measure the kinetics of L1 retrotransposition in cultured cells. The GloMax® Multi Luminometer was used to quantify luciferase activity. (4205)
GloMax®-Multi+ Luminescence Module
pGL4.13[luc2/SV40] Vector
pGL4.73[hRluc/SV40] Vector
Li, Y., Song, M., Kiledjian, M.
Notes: This study analysed the roles of Dcp2 and Nudt16 in nonsense-mediated mRNA decay miRNA-mediated silencing. The authors used various luciferase reporter constructs to evaluate the significance of Dcp2 and Nudt16 in miRNA- and siRNA-mediated gene silencing in wildtype and knockout MEF cells. Various Renilla luciferase constructs containing or lacking miRNA target sites were cotransfected along with a control plasmid encoding firefly luciferase (for normalization purposes). Renilla and firefly luciferase luminescence were measured using the Dual-Luciferase® Reporter Assay and the GloMax®-Multi Luminometer. (4204)
Kweider, N., Fragoulis, A., Rosen, C., Pecks, U., Rath, W., Pufe, T., and Wruck, C.J.
Notes: These authors investigated the relationship between VEGF and oxidative stress related to preeclampsia. They showed that VEGF activates Nrf2 in an ERK1/2-dependent manner, protecting against oxidative stress. They first used a dual-luciferase reporter assay and a pGL3-ARE vector construct to show that VEGF activates ARE in the cytotrophic cell line BeWo. Firefly and Renilla luciferase activities were determined using the Dual-Luciferase® reporter assay system and the GloMax®-96 microplate luminometer. The authors then showed that inactivation of the transcription factor Nrf2 by shRNA abolished this VEGF-dependent ARE activation. To determine whether Nrf2 protected BeWo cells from oxidative stress, cells were pretreated with VEGF and then exposed to H2O2 before monitoring cell viability and cytotoxicity. Cytotoxicity assays were performed using the CytoTox-Glo™ Assay. (4199)
GloMax® 96 Microplate Luminometer
CytoTox-Glo™ Cytotoxicity Assay
Racca A.C., Camolotto S.A., Ridano M.E., Bocco J.L., Genti-Raimondi S., and Panzetta-Dutari, G.M.
Notes: These authors studied KLF6 expression during human trophoblast cell differentiation, and its role in the regulation of genes associated with placental development and pregnancy maintenance. They used immunofluorescence microscopy, RT-qPCR and luciferase reporter assays to investigate cellular localization, mRNA expression, and transcriptional activation. Reporter assays were performed using various luciferase reporter constructs, the Dual-Luciferase® Assay, and the GloMax®-Multi Detection System. KLF6 was shown to play a role as transcriptional regulator of relevant genes for placental differentiation and physiology such as βhCG and PSG. (4197)
Passive Lysis 5X Buffer
GloMax®-Multi Base Instrument
Shin, O.S., Uddin, T., Citorik, R., Wang, J.P., Della Pelle, P., Kradin, R.L., Bingle C.D., Bingle, L., Camilli, A., Bhuiyan, T.R., Shirin, T., Ryan, E.T., Calderwood, S.B., Finberg, R.W., Qadri, F., Larocque, R.C. and Harris, J.B.
Notes: The authors used the Dual-Glo™ Luciferase Assay system to detect induction of NF-κB activity in transfected HEK-TLR4/MD2 cells expressing human CD14, toll-like receptor 4 (TLR4) and MD2. An NF-κB-driven luciferase reporter gene and a thymidine kinase (TK) promoter-driven Renilla luciferase reporter gene were measured 24 hours after the cells were exposed to long palate, lung, and nasal epithelium clone 1 protein (LPLUNC1) 1 hour prior to stimulation with lipopolysaccharide (LPS) from either E. coli or V. cholera. Figure 4 shows how LPLUNC1 was able to inhibit the TLR4 response to LPS from V. cholera. It was hypothesized that LPLUNC1 serves to reduce inflammation to enteric pathogens. Transfection details: Cells were seeded in 96-well plates at 30,000 cells/well and transfected with a total of 0.3µg of DNA per well. The transfected DNA contained 80ng of NF-κB-firefly luciferase plasmid and 20ng HSV-TK promoter-driven Renilla luciferase plasmid, along with human CD14 construct cloned into pCDNA3 at 10ng/well. (4177)
Jheon, A.H., Mostowfi, P., Snead, M.L., Ihrie, R.A., Sone, E., Pramparo, T., Attardi, L.D. and Klein, O.D.
Notes: The authors determined that PERP, a tetraspan membrane protein, is required for enamel formation during tooth development in mice. Using microarray analysis, they then identified genes that were differentially expressed in wildtype and Perp-null mice and might be involved in enamel formation. Differential expression of these genes was confirmed by qPCR using the GoTaq® qPCR Master Mix. The authors also identified an upstream regulator of Perp, P63, by transfecting cells derived from embryonic mouse teeth with a Perp-luciferase reporter construct that contained a P63 response element or mutated P63 response element. A Renilla luciferase vector was used for normalization of transfection efficiency, and luciferase assays were performed using the Dual-Luciferase® Reporter Assay System. (4168)
GoTaq® qPCR Master Mix
Stempin, C.C., Chi, L., Giraldo-Vela, J.P., High, A.A., Häcker, H. and Redecke, V.
Notes: The authors used the Dual-Luciferase® Reporter Assay System to measure NF-κB firefly luciferase activity normalized to Renilla luciferase (pRL-TK) in transfected HEK293T and EL4 cells. Coumermycin-treated HEK293 and EL4 cells transiently transfected with B-cell CLL/lymphoma 10 (BCL10) led to transcriptional NF-κB activation in a dose dependent manner. Dimerization of BCL10 by coumermycin was used to mimic physiological stimulation through T cell receptor cross-linking, which initiates BCL10-mediated activation of the NF-κB signaling pathway. Overexpression experiments showed that E3 Ubiquitin Ligase Mind Bomb-2 (MIB2) controlled BCL10-mediated activation of NF-κB by promoting autoubiquitination and ubiquitination of IKKγ, a kinase responsible for phosphorylating IκBα protein. The authors identified that the C-terminal RING finger domain of MIB2 was critical for protein ubiquitination in NF-κB activation, which was confirmed by the NF-κB luciferase reporter response to various MIB2 mutants. (4180)
Maruyama, A., Nishikawa, K., Kawatani, Y., Mimura, J., Hosoya, T., Harada, N., Yamamato, M. and Itoh, K.
Notes: These authors first used a FLAG-tagged protein (nfr2) with a HeLa Nuclear extract and captured interacting proteins via SDS-PAGE and in-gel digests of bands to identify (Krüppel-associated box)-associated protein 1 (KAP1) as a potential interacting partner. Human KAP1 was purchased as a HaloTag® CMV Flexi® Vector from Kazusa and used in a Mammalian PullDown scenario (with HaloLink™ Resin) to demonstrate interaction between the two proteins. A reporter assay was used to show that KAP1 facilitates Nrf2 transactivation in a dose-dependent manner. The authors defined the interaction sites using GST-tagged nrf2 and various forms of KAP1-HaloTag® Fusions expressed in TNT® SP6 High-Yield Wheat Germ Extract. GST-tagged proteins were expressed in E. coli and bound to glutathione-Sepharose beads. These bound proteins were mixed with the KAP1 from the cell-free expression system, incubated for 4 hours at 4°C, washed and stained with the HaloTag® TMR Ligand for 30 minutes. The proteins from the pull-down assay were subjected to SDS-PAGE and the HaloTag® proteins detected by phosphorimaging and the GST proteins by Coomassie Brilliant Blue Staining. A two-hybrid system consisting of the pRL-TK Vector with a firefly luciferase reporter with Gal4 UAS, mouse Nrf-2 N-terminal domain and KAP1 was also used. The vectors were transfected into Nrf2 knockout MEFs for 4 hours then incubated for 36 hours before luciferase expression was determined using the Dual-Luciferase® Reporter Assay System. (4123)
Anti-HaloTag® pAb
HaloTag® TMR Ligand
pFN21A HaloTag® CMV Flexi® Vector
Galatioto, J., Mascareno, E., and Siddiqui, M.A.
Notes: These authors investigated the role of the cardiac lineage protein 1 (CLP1/HEXIM1) in skeletal myogenesis. They showed that CLP1 knockout C2C12 cells were unable to differentiate, and then investigated the hypothesis that CLP1 associates with MyoD and HDAC proteins to downregulate cell cycle genes, such as cyclin D1, and allow expression of differentiation-specific genes. RNasin® Ribonuclease inhibitor was used in coimmunoprecipitation assays investigating the interaction between CLP1 and HDAC in C2C12 cells under differentiation and non-differentiation culture conditions. RNasin® was included during cell lysate preparation prior to coimmunoprecipitation assays with antibodies directed against various HDAC proteins. The authors also performed a luciferase reporter assay using the Dual-Luciferase® Assay System to investigate the regulation of cyclin D1 expression by CLP1, MyoD and HDAC. A luciferase reporter-cyclin D1 construct was co-transfected with MyoD, HDAC and CLP1 constructs and the effect on luciferase expression examined under differentiation and non-differentiation conditions. In differentiation medium MyoD, CLP1 and HDAC5 acted synergistically to reduce cyclin D1 expression. (4225)
Recombinant RNasin® Ribonuclease Inhibitor
Meloni, A., Fiorillo, E., Corda, D., Incani, F., Serra, M.L., Contini, A., Cao, A. and Rosatelli, M.C.
Notes: The authors used a yeast two-hybrid system to identify proteins that interact with the autoimmune regulator (AIRE) protein. DAXX, a multifunctional protein involved in apoptosis and transcription regulation, interacts with AIRE, as shown through coimmunoprecipitation and colocalization studies. Colocalization of AIRE and DAXX in HeLa cells was demonstrated by confocal microscopy using a Monster Green® Fluorescent Protein-AIRE fusion protein and endogenous DAXX, which was detected using an anti-DAXX primary antibody and an anti-rabbit secondary antibody conjugated with Texas Red fluorophore. However, AIRE and DAXX did not interact in vitro in a GST pull-down assay using a GST-AIRE construct, radiolabeled DAXX protein expressed in a TNT® system, and MagneGST™ Glutathione Particles, leading the authors to speculate that the interaction is weak or there are scaffold proteins required for protein interaction. To examine the effect of DAXX on AIRE transcriptional activity, the authors transfected COS-1 and HeLa cells with AIRE and DAXX expression constructs and a luciferase reporter plasmid with the human insulin promoter, then performed Dual Luciferase® Reporter Assays. AIRE induced transcription of the insulin promoter, but coexpression of DAXX suppressed this transcriptional activation. (4153)
MagneGST™ Glutathione Particles
Monster Green® Fluorescent Protein phMGFP Vector
Guo, H., Mi, Z., Bowles, D.E., Bhattacharya, S.D. and Kuo, P.C.
Notes: The authors investigated the role of the ubiquitin E3 ligase PDLIM2 in the degradation of signal tranducer and activator of transcription 1 (STAT1). They showed that activation of PDLIM2 and subsequent STAT1 ubiquitination require protein kinase C-mediated phoshorylation of PDLIM2 on serine 137. Polyhistidine-tagged PDLIM2 and polyhistidine-tagged mutants PDLIM2-S137A and PDLIM2-S137D were purified using the MagneHis™ Protein Purification System for use in in vitro phosphorylation and ubiquitination assays. One mechanism used to assess levels of STAT1 was a reporter assay using RAW264.7 cells transfected with a pGL3-based construct containing a interferon γ-activated sequence (GAS) upstream of the firefly luciferase reporter gene. Expression of wildtype PDLIM2, but not the mutant forms, resulted in much lower levels of STAT1 protein, and thus lower luciferase activity, when cells were challenged with lipopolysaccharide. The pRL-TK Vector was used to normalize for transfection efficiency. Luciferase assays were performed using the Dual Luciferase® Reporter Assay System and Reporter Lysis Buffer. (4152)
MagneHis™ Protein Purification System
Hornakova, T., Staerk, J., Royer, Y., Flex, E., Tartaglia, M., Constantinescu, S.N., Knoops, L. and Renauld, J.C.
Notes: The authors studied the ability of JAK1 V658F and A634D mutants to activate the Janus kinase (JAK)/STAT pathway when expressed alone or together with the other components of the interleukin-9 receptor complex. The BOX1 motif of wild-type IL-9Rα, the JAK interacting region, was mutated from PXP to SXS using the GeneEditor™ in vitro Site-Directed Mutagenesis System. To assess STAT transcriptional activity, HEK293 human embryonic kidney, COS-7 monkey kidney, U4C human fibrosarcoma and g2A cells were cotransfected with 250ng of the appropriate constructs, 500ng of firefly luciferase vectors and 50ng of pRL-TK Vector and empty plasmid for a total 1.5µg of DNA. After 24 hours, the cells were lysed in 150µl of Passive Lysis Buffer and reporter activity measured using the Dual-Luciferase® Reporter Assay System. The ProFection® Mammalian Transfection System—Calcium Phosphate was used to transfect 106 HEK293 cells in a six-well plate with 3.75µg of plasmid for Western blot analysis and cotransfected 6 × 106 HEK293 cells in a 100mm dish with 14µg plasmid for immunoprecipitation studies. (4025)
ProFection® Mammalian Transfection System-Calcium Phosphate
Cho, Y.Y., Tang, F., Yao, K., Lu, C., Zhu, F., Zheng, D., Pugliese, A., Bode, A.M. and Dong, Z.
Notes: This paper examined the role of the Cdk3/c-Jun signaling in EGF-stimulated cell proliferation and cell transformation. An AP-1 luciferase reporter plasmid construct was contransfected with various expression vector combinations of Cdk3, Cdk2, c-Jun, c-Jun M63/73, Cdk3-DN and cyclin C and the phRL-SV40 Vector as a normalization control. Cells were lysed at room temperature for 30 minutes with gentle shaking and luciferase activity measured using the Dual-Luciferase® Reporter Assay System. SaoS-2 cells transfected with a mock and Cdk3 RNAi vector were seeded at a density of 4 x 103 in 96-well plates with 20µl of medium. After 24 hours, 20µl of CellTiter® 96 AQueous One Solution was dispensed to each well, and the plate incubated for 1 hour at 37°C. The reaction was stopped using 25µl of 10% SDS and absorbance was measured at 492 and 690nm. For the CheckMate™ Mammalian Two-Hybrid System, the plasmid constructs, pACT-c-Jun, pBIND-Cdk3 and pG5luc, were cotransfected into HEK293 cells at no more than 100ng/well in a 48-well plate. After incubation and lysis, 20µl of lysate was dispensed into each well of a 96-well luminescence plate. Firefly and Renilla luciferase activity was detected. (4028)
CellTiter 96® AQueous One Solution Cell Proliferation Assay
CheckMate™ Mammalian Two-Hybrid System
Stros, M., Polanská, E., Struncová, S. and Pospísilová, S.
Notes: The authors examined whether HMGB1 and HMGB2 proteins could affect promoter activity of the topoisomerase IIα gene. Portions of the topoisomerase IIα gene promoter were cloned into the pGL3 Basic Vector, and Saos-2 cells were cotransfected with the resulting constructs, an HMGB1- or HMGB2-expressing plasmid and the pRL-tk Vector as a control for normalization. Firefly and Renilla luciferase activities were determined using the Dual-Luciferase Reporter Assay. To determine whether HMGB1 and HMGB2 promoted binding of the transcription factor nuclear factor-Y (NF-Y) to the topoisomerase IIα promoter, the authors used a chromatin immunoprecipitation (ChIP) assay. Two populations of Saos-2 cells, one of which expressed HMGB1 or HMGB2 and one that had expression inhibited, were fixed with formaldehyde, then treated to shear chromatin. Immunoprecipitation was performed using an anti-NF-Y antibody, and the amount of DNA bound to the NF-Y was quantified by semi-quantitative PCR using GoTaq® Hot Start DNA Polymerase and Green GoTaq® Flexi Reaction Buffer. (4037)
GoTaq® Hot Start Polymerase
T4 RNA Ligase
Shigemoto, T., Kageyama, M., Hirai, R., Zheng, J., Yoneyama, M. and Fujita, T.
Notes: Here the authors studied various non-synonymous SNPs of retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5) that are essential for detecting viral RNA and triggering antiviral responses. Various point mutations were introduced into RIG-1 and MDA5 using the GeneEditor™ in vitro Site-Directed Mutagenesis System with pEF-FLAG clones. Mouse embryonic fibroblasts (MEFs) and L929 cells were cotransfected with RIG-I mutants or MDA5mutants and pRL-TK Vector, and stimulated with RNA or viral infection. Reporter activity was measured using the Dual-Luciferase® Reporter Assay System. (4024)
Wilson, P.M., Fazzone, W., LaBonte, M.J., Lenz, H.J. and Ladner, R.D.
Notes: The authors examined the role of p53 in modulating dUTPase promoter activity. Base substitution mutations of Sp1- and E2F-binding sites in the dUTPase promoter were performed using the GeneEditor™ in vitro Site-Directed Mutagenesis System. Each mutant was confirmed by DNA sequencing. To determine growth inhibition, HCT116 human colon cancer cells were seeded in 96-well plates at 3 × 103 cells/well and treated with 5-fluorouracil (5-FU), fluorodeoxyuridine (FUdR), oxaliplatin or in combination. After 72 hours, the CellTiter® 96 AQueous One Solution was dispensed into each well and absorbance measured. RNA was isolated from HCT116 p53+/+ and HCT116 p53-/- cells. cDNA was reverse transcribed from 200ng total RNA followed by multiplex qPCR using the Plexor™ qPCR System to amplify dUTPase, thymidylate synthase and GAPDH, a housekeeping gene. The 1.2 kb region of the dUTPase promoter upstream of the transcriptional start site was amplified by PCR and the fragment cloned into the pGL3-Basic Vector. Truncated promoters were also generated by PCR and cloned into the same vector. Drosophila SL-2 cells and HCT116 cell lines were seeded in a 24-well plate and transfected with dUTPase pGL3 promoter constructs or with pCI-Neo:p53WT, pCI-Neo:p53MUT and the empty pCI-neo Mammalian Expression Vector; all transfections included the pRL-TK Vector at a ratio of 1:10. After six hours, the cells were incubated in either fresh medium or medium containing a cytotoxic agent at the appropriate concentration. Thirty hours later, the cells were lysed, quantitated by Western blotting and 20µl of lysate analyzed with the Dual-Luciferase® Reporter Assay System. Electrophoretic mobility shift analyses (EMSA) were performed using –64 to –91 of the dUTPase-nuclear isoform transcriptional start site in the Gel Shift Assay System. (4031)
Gel Shift Assay Core System
Gel Shift Assay System
pCI Mammalian Expression Vector
Plexor® qPCR System
Rahmani, F., Hummel, M., Schuurmans, J., Wiese-Klinkenberg, A., Smeekens, S. and Hanson, J.
Notes: The authors were wanted to study the upstream open reading frame 2 (uORF2) of the 5' leader of bZIP11 mRNA, which has a role in sucrose regulation. The whole 5' leader fragment of bZIP11 was subcloned into the pALTER® Vector and amino acid substitutions were introduced using the Altered Sites® II in vitro Mutagenesis System. The pGEM®-T Easy Vector was used to clone two PCR fragments that were then subcloned using restriction enzymes to create a fusion of uORF2 to a different 5' leader. Arabidopsis seedlings were transformed via particle bombardment. 20mg of plant tissue was ground in Passive Lysis Buffer, centrifuged, and 20µl of the supernatant was assessed for reporter gene expression using the Dual-Luciferase® Reporter Assay System. (4023)
pGEM®-T Vector System I
pGEM®-T Vector System II
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If you make one of these for your house or classroom I would love to hear about it!
Pink and Green Mama,
Love, love, love this idea - my twins would have a blast, I will have to make them one.
And thanks for popping over to my blog to enter my giveaway!
Such a great idea!
This is such a great idea. I know my son would love the construction one! I'm going to have to try this myself.
That's a great idea! Very nice of you to brainstorm ideas with the kindergarten teacher too.
I grew a little pumpkin in our backyard once when I was a kid. Just planted some seeds one year, and one actually grew! If I remember correctly, someone stole it.
What a GREAT idea!!!!! OMGosh, I am so getting a tub, and rocks for my little boys. They spend hours outside playing in the dirt, and I never though of bringing the outside fun in - that would so help on rainy, or yucky snowy days. Thanks for the great idea! ;)
Sex Diaries of a Mom
I love your idea so much I'm going to borrow it for my boys. Thanks
What a cool idea!
Omg what a great idea! I have a crazy fun little 3 year old who would just love such a thing! Thanks for sharing.
This is so much fun! I just got throught making a Construction Box for my classroom with dump trucks we got at a garage sale! What a deal!
My DS loves Bob the Builder too and has some of the same figures! We were just playing with them a few minutes ago... I love this idea with the stones. If I can keep the 1-yr-old away, I'll try it! :)
Oh, and pleeease tell me where you got that delicious rug!! I LOVE it! Perfect for a fun, colorful playroom!!!
Oh, and I just can't stop looking, so you have a new follower... ;)
I stumbled across your blog through Playopedia. Found this activity and had to make it for my daughter. She's absolutely enthralled with it. Thank you so much for a great idea! :)
You have done it again! Kiddos love the Ocean book and will go bonkers for Under Construction. THANK YOU!!
This is a really cool idea. I think I'm going to try something like it in my classroom. Thanks for sharing!
Heather Mix
An Interesting Mix
Love this!! I will have to use this just for construction site toys! We live on a farm and I have done with this with oatmeal and rice and used farm tractors, wagons, augers, etc!! Thanks so much for sharing and what a neat blog!!
What a fantastic idea - I really look forward to checking out lots more of your posts. Finding you on Pinterest definitely makes the the five hours I spent on there worth it... LOL
I just saw this on Pinterest. I had totally forgotten this idea for my little guy - so happy for the reminder with winter on the way!
I teach Pre-K and my 4-5 year old boys would love this! I am definitely going to have to make one of these take it in! Thanks so much for sharing :)
I would love to do this for my nephew who LOVES trucks and big machinery. Where did you find the small trucks and dozers and things? I have looked everywhere and can't find any small ones like that.
Hi Donna, the construction set was a generic set from Walmart years ago (maybe 7 years ago?) at Christmas time. Check there or at Toys R' Us in their generic toy areas for a set!
Can you please reply with the dimensions of your storage box?
Margaret Mary,
We use plastic storage boxes for all of our sensory boxes. Approximately 12" wide x 24" long ' 3"-4" high.
I LOVE this idea--can't wait to try it with my son! And I love that your girls are involved. I feel like our society pushes boys to play with bulldozers and girls to play with princess dolls, and it drives me crazy! Thanks for being part of the solution.
These posts never get old ... so much fun! I featured your post and photo in my Montessori-Inspired Construction Unit at http://livingmontessorinow.com/2014/05/28/montessori-inspired-construction-unit/
Disclosure of Material Connection: My family and I received one or more of the products or services mentioned in some of my posts for free in the hope that I would mention it on my blog. I only recommend products or services I use personally and believe my readers will enjoy in connection with my blog theme. I am disclosing this in accordance with the Federal Trade Commission's 16 CFR, Part 255: "Guides Concerning the Use of Endorsements and Testimonials in Advertising." All opinions expressed on this blog are my own and all content is mine. I do not post pre-written posts from outside parties. My lesson plans and craft ideas are not to be sold for profit by others. Please play nicely. This blog does not share personal information with third parties nor do we store any information about your visit to this blog other than to analyze and optimize your content and reading experience through the use of cookies. You can turn off the use of cookies at anytime by changing your specific browser settings. We are not responsible for republished content from this blog on other blogs or websites without our permission. This privacy policy is subject to change without notice and was last updated on April 21, 2013. Pink and Green Mama is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to amazon.com.

subset: commoncrawl


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NORTON, Mass. (AP) — The question was inevitable and the answer was predictable. Tiger Woods says his back is fine.
Woods played in the pro-am Thursday morning at the Deutsche Bank Championship. The last image of Woods on the golf course looked much worse. Four days ago, he dropped to his knees after a back spasm during the final round of The Barclays at Liberty National, where he finished one shot behind Adam Scott.
Woods said after the round it was "hypothetical" whether he would play the Deutsche Bank, adding that he had just walked off the course and wasn't feeling very good.
One of his amateur partners asked him about his back on the first tee at the TPC Boston. Woods gave a short answer.


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By Mike Rush
Maryland Heights, MO (KSDK) - "A little bit of a bummer" was served up Wednesday morning with the burgers and beers at the restaurant Pujols 5, co-owner Patrick Hanon said.
By the time the not-so-well kept secret became official at the restaurant, Cardinals fans could not hide their disappointment.
"I think everybody's a little concerned," said fan Lisa Bulczak.
Because the announcement means no hit, not even a strikeout, in the ongoing contract inning. The fans and the restaurant were ready for good news.
"We've got champaign on ice for if and when he does make the announcement," Hanon said.
But there was no bubbly and who knows if it will ever be needed.
"Now we just have to deal with it looming over us all year and go from there," said Cardinals fan Jason Thomas.
Some fans think the time could be an eye-opener for Pujols and help him realize what he has in St. Louis. But there are other theories.
"I think if you put more time between the deal, I think it's going to give him more time to think. I think it may be the deal breaker," said fan John Reynolds, who was also in the restaurant when the official news was announced on the big screen television sets.
Whether he's gone or not, Pujols has at least one more season and these fans don't plan on holding a grudge.
"Cardinals fans are Cardinals fans and I think they're here to watch baseball and why wouldn't you cheer for the best player in the game," said fan Glenn Flavin.


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7 Possible Replacements for Timothy Geithner
News reports said Treasury Secretary Geithner wanted to leave his post once debt negotiations were complete. With that milestone past, speculation now turns toward potential successors. The stakes are high, with unemployment at 9.2% and partisan theatrics as melodramatic as ever. Here are seven candidates that the president might do well to consider.


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When you take a South Africa tour with Grand Circle Travel, you'll discover cultural and natural wonders at a relaxed and leisurely pace. You'll follow exotic wildlife on game-viewing drives through Kruger National Park and the Hluhluwe-Umfolozi Game Reserve ... enjoy visits to charming coastal towns along the famed Garden Route ... and explore cosmopolitan Cape Town, South Africa's "Mother City." You'll marvel at the country's diverse landscape rife with verdant, grassy savannahs, balmy, white-sand beaches, and tranquil, flower-ringed lagoons. And everywhere you go, you'll connect with the people of South Africa, encountering a veritable kaleidoscope of cultures and creeds—evidence that the nation has indeed become, as Nelson Mandela envisioned, "a rainbow nation at peace with itself and the world."
hiddenDay 1Depart U.S.
Board your overnight flight to Johannesburg, South Africa.
hiddenDay 2Arrive Johannesburg, South Africa
A Grand Circle representative will meet you at the airport and help you transfer to your hotel, where you'll meet the Grand Circle Program Director who will accompany you throughout your trip, and fellow travelers, including those who have just finished our optional pre-trip extension, Big Five Bush Safari, South Africa. Settle in, and get to know your surroundings.
hiddenDay 3Johannesburg • Soweto tour • Apartheid Museum
This morning, you'll drive to Soweto, enjoying scenic views of Johannesburg en route. You'll see the city skyscrapers, including the Carlton Centre, which is the tallest office block in Africa, as well as the Nelson Mandela Bridge.
Soweto, a former shantytown on the city's outskirts, is both a bustling center of black South African life and a poignant symbol of the struggle to end apartheid. Here, after an included lunch, you'll set off on a walking tour, where you'll enjoy the opportunity to immerse yourself in local culture as you connect closely with the township's residents, visiting them in their homes and where they work.
After your walking tour, depart for the Apartheid Museum, where you'll confront one of the 20th century's greatest injustices. The history of nearly 50 years of unequal rights affecting more than 20 million black South Africans is recounted here, in 22 exhibits spanning print, photo, and film. It's a fascinating and moving examination of an era that has since been replaced by a constitutional democracy that celebrates diversity.
This evening, get to know your fellow travelers during a Welcome Briefing, and enjoy a Welcome Dinner with wine at your hotel.
hiddenDay 4Johannesburg • Mpumalanga • Kruger National Park
After breakfast, set off for Kruger National Park. Along the way, marvel at breathtaking views of the Klein Drakensburg escarpment, with its densely wooded canyons, sheer sandstone cliffs, and sparkling waterfalls. You'll stop for lunch on your own en route to your hotel in the province of Mpumalanga.
You'll travel the Panorama Route through Mpumalanga, where you'll look into God's Window—a spot with stunning views—and the Bourkes Luck Potholes at the mouth of the Blyde River Canyon.
Tonight, check into your hotel, where dinner will be served.
hiddenDay 5Kruger National Park • Intimate African Safari game drive
Embark on a full-day Intimate African Safari game drive, during which you will explore Kruger National Park aboard a small, open-air vehicle. Perhaps you'll see one of the elusive "Big Five" safari animals during your journey. A packed lunch will be provided by the hotel.
Kruger is one of the largest and most notable among African game parks. Transvaal President Paul Kruger saw the need to protect the wilderness and its animals and first proclaimed it the Sabie Game Reserve before it became a national park in 1926. It is known for the chances it offers for spotting the Big Five—always elusive, but certainly present here: lion, elephant, leopard, Cape buffalo, and rhino. In addition to game animals, Kruger boasts an avian population of almost 500 species, some of which are found nowhere else in South Africa.
Tonight, enjoy dinner at your hotel.
hiddenDay 6Kruger National Park • Full-day game drive • Optional Intimate African Safari
After breakfast, you'll board a coach for a full-day game-viewing drive.
Or, choose to take advantage of a second Intimate African Safari on our optional full-day outing. Today, you'll venture further afield to some of the park's most remote areas aboard 4x4 vehicles to discover South Africa's wildlife in a smaller group. Please note: A minimum of six participants is required to operate this tour.
Dine tonight at your hotel.
hiddenDay 7Kruger National Park • School visit • Mbabane, Swaziland
This morning, spend time with the children of George Mhaule Primary School (when in session), supported by Grand Circle Foundation as part of the World Classroom initiative. After your visit, cross the border into the Kingdom of Swaziland, entering through its mountainous northwest. As you travel, admire a landscape of sloping hills and sugarcane plantations.
A fully independent nation since 1968, this African kingdom originated in the early 19th century and maintained its cultural identity as a British protectorate—never absorbed into South Africa—through much of the 20th century. Today, Swaziland has a population of about 800,000 and is a peaceful, agricultural country where ancient and modern ways mingle. Arrive at your hotel, near the capital city of Mbabane, in the mid-afternoon with a bit of time to explore the area or relax before dinner.
hiddenDay 8Mbabane • Swazi Cultural ExperienceFOUNDATION VISIT
Today you'll enjoy a full day of a Swazi Cultural Experience. First, peruse the produce, local crafts, and wares of the Swazi people at the Mbabane Craft Market. Continue on to a local workshop to observe the artistry and precise techniques of glassblowing.
Then, behold the craftsmanship of local candle-makers as they work their magic in wax—using an age-old process—at a local workshop and market. Enjoy lunch here before making a short stop at the memorial built in honor of King Sobuza, who brought Swaziland to independence in 1968. Crowned king at less than a year old, King Sobuza holds the record for the longest monarchial rule—more than 80 years.
This evening, dinner is on your own—your Program Director can offer suggestions about where you can savor the local flavors.
hiddenDay 9Mbabane • Hluhluwe, South Africa • Afternoon With the Zulu
As you cross the border back into South Africa today, you'll enter Hluhluwe, a small town within the Kwazulu-Natal province. Here the great warrior-king Shaka unified many clans in the early 19th century under the name Zulu, which was the name of his own clan. Though mostly ceremonial today, the Zulu monarchy continues as a proud symbol of a living culture.
While in Hluhluwe, you'll spend an Afternoon With the Zulu during an exclusive Discovery Series event. You'll visit an authentic re-creation of a Zulu uzumi (homestead), where you'll meet modern-day warriors and witness fighting formations. You'll also enjoy lunch, take part in traditional activities such as beer-brewing and bread-making, and discover some of the secrets of the mystical nyangas (herbal healers) and the Sangoma (a priest-diviner).
Tonight, dinner will be served at the hotel.
hiddenDay 10Hluhluwe • Hluhluwe-Umfolozi Game Reserve • Optional Hluhluwe-Umfolozi afternoon game-viewing drive
After breakfast, set off for a game-viewing drive through the Hluhluwe-Umfolozi Game Reserve—the oldest park in all of Africa.
Thanks to the efforts of conservationists, the once-threatened rhino population has soared—the park is now home to the largest concentration of white rhinos in the world. Be on the lookout for their signature horns, along with spottings of the "Big Five," and the more obscure sightings of yellow-, pink- and orange-throated longclaws.
Later, after lunch on your own, you can spend the rest of the day at leisure. Or, if you're eager for another look at the exciting wildlife of the Hluhluwe-Umfolozi Game Reserve, you can opt to join a second tour in the afternoon. You'll enjoy another opportunity to seek out the residents of the reserve, seeing the animals in a new light at a different time of day.
Tonight you'll return to your hotel for some leisure time before dinner.
hiddenDay 11Hluhluwe • Overland to Durban • Fly to Port Elizabeth • Port Elizabeth City tour
This morning, after an early wake-up, you'll drive to Durban. From here, you'll fly to Port Elizabeth—a major seaport set along the beautiful shores of Algoa Bay.
Enjoy a tour of the city's historical attractions, including St. George's Park and Settler's Park—both known for their cultivated and natural gardens. The afternoon is at leisure for you to explore the shimmering beaches and colonial history of "the Friendly City." This evening is at leisure.
hiddenDay 12Port Elizabeth • Optional Addo Elephant National Park tour
Spend the day and evening as you please in Port Elizabeth. The city has many historical attractions—including the Donkin Heritage Trail, which allows visitors to follow in the footsteps of the British settlers who arrived in 1820. The trail, which is several miles long, winds past more than 40 historical sites and architectural gems in the Old Hill area of central Port Elizabeth. You might also choose to stroll through St. George's Park, home to the world-famous Port Elizabeth Cricket Club. Lunch and dinner today are on your own.
Or join an optional motorcoach tour of Addo Elephant National Park, where you'll enjoy another chance to see South Africa's most celebrated safari animals. The park houses more than 450 elephants, 400 Cape buffalo, nearly 50 endangered black rhino, and a variety of antelope. You may even see lion and spotted hyena, which have been re-introduced to the area in recent years.
hiddenDay 13Port Elizabeth • Garden Route to Knysna
After breakfast this morning, you'll embark on a scenic journey along South Africa's famous Garden Route, the magnificent stretch of coastline that has served as an inspiration for artists and writers for centuries. View lofty mountains edging toward some of the best golden-sand beaches in the country ... vast forests of giant yellowwood, pine, and eucalyptus trees ... colorful wildflowers ... pristine, amber-colored lakes, and tumbling waterfalls ... coves and lagoons peeking out from behind a ridge of sand dunes ... and charming tiny villages.
This evening, you'll arrive in Knysna, known as the "Pearl of the Garden Route." Dinner is yours to enjoy at a local restaurant tonight.
hiddenDay 14Knysna • Oudtshoorn Ostrich Farm
Today, get closer to the world of exotic animals in Oudtshoorn during a visit to the Safari Ostrich Farm.
Ostrich farming began in the late 1880s, when European demand for ostrich feathers prompted a boom in ostrich farming worldwide. By the early 1900s, only gold, diamonds, and wool ranked higher than ostrich feathers in terms of exports. In Oudtshoorn, the center of the industry, ostrich farmers built lavish sandstone mansions to showcase their wealth.
On the farm you'll view large flocks of the flightless bird, still highly prized for its feathers, leather, and meat. Experienced guides will reveal the life cycle of the ostrich, and you'll also learn how products such as handbags and feather dusters are produced. Perhaps you'll even observe and participate in an ostrich derby! After lunch at the farm, you'll have more time at leisure to explore Oudtshoorn.
You'll return to Knysna tonight in time for dinner on your own.
hiddenDay 15Knysna • Featherbed Reserve Nature Walk
After breakfast, depart for Knysna's famed lagoon, which is protected from the open water by two giant sandstone cliffs known locally as "the Heads." A ferry takes you across the lagoon for an included tour of Featherbed Nature Reserve. On your guided nature walk, keep an eye out for a glimpse of the blue duiker, Africa's smallest antelope (named for the blue sheen of its back) and other wildlife.
After lunch outside under the reserve's milkwood trees, the remainder of the day is yours to explore Knysna. You can delve into the town's history at the Millwood House Museum, which displays items once owned by Knysna's founder, George Rex (reputedly an illegitimate son of British King George III). Or browse through the many quaint shops with their unique offerings.
Dinner tonight is on your own.
hiddenDay 16Knysna • Swellendam • Cape Town
Ride to Cape Town after breakfast this morning. En route, you'll stop in historic Mossel Bay, where Portuguese explorer Bartolomeu Dias became the first European to land in South Africa in 1488. For more than 400 years, this was a spot where passing mariners dropped off their letters as their voyages took them around the Cape of Good Hope. You'll learn more about the legacy of this explorer, as well as about the culture and ecosystem of Mossel Bay, at the eponymous Dias Museum on an included tour.
You'll also stop at the town of Swellendam, where you'll enjoy lunch at a local restaurant, before continuing on to Cape Town, where you'll arrive this evening. Enjoy dinner on your own tonight.
hiddenDay 17Cape Town • Cape Town City tour • Optional Cape Town Townships tour • Home-Hosted Dinner
Today's included city tour features a cable car ride up Table Mountain (weather permitting), a 3,563-foot sandstone mountain that is remarkable for its flat top. Enjoy the ocean views from the summit.
This afternoon, enjoy leisure time to explore Cape Town on your own.
Or perhaps you'll join our optional tour to Cape Town's vibrant townships—Langa, Guguletu, and Bonteheuwel. Learn about the traditions and daily way of life here through a tour—guided by a township resident—and a home visit.
Tonight, join a local family in their home for dinner. Enjoy a warm welcome, authentic local fare, and lively conversation during this exclusive Discovery Series event.
hiddenDay 18Cape Town • Cape Peninsula
Departing after breakfast, head along the coast, with majestic Table Mountain rising in the distance. Your route follows the spectacular coast road, the Atlantic Ocean's crashing surf endlessly splashing the rocks below. Soon you reach the Cape Point Nature Reserve, with its wild fynbos landscapes, beautiful flowers, elusive bontebok, baboons, and ocean views.
Contrary to popular belief, the Cape Peninsula is not consistently where the Indian and Atlantic oceans meet. Because of shifting currents, that distinction is shared with lesser-known Cape Agulhas, 100 to 200 miles east of the Cape Peninsula. Nevertheless, when you get to the tip of the Cape Point Nature Reserve, you'll see Cape Point, the technical "Cape of Good Hope." Rapidly changing climactic conditions and the Indian Ocean currents coming from Cape Agulhas make it a particularly dangerous spot for ships. Your Program Director will tell you that more "sightings" of the legendary ghost ship, the Flying Dutchman, are reported here than anywhere else in the world. Who knows what you'll spy on the distant horizons as you gaze from the Cape Point viewing platform? You'll also explore some of the fascinating vegetation and more remote corners of this nature reserve, as well as the lush Kirstenbosch Botanical Gardens, before stopping for an included lunch at a well-known local seafood restaurant.
On the return trip to Cape Town, a stop along the False Bay coastline offers a chance to see the penguins at Boulders Beach. Arrive back in town in the early evening, after passing through Simon's Town and Fish Hoek.
Dinner is on your own tonight.
hiddenDay 19Cape Town • Stellenbosch
Venture to South Africa's wine-producing region on an included excursion to Stellenbosch. You'll visit a wine estate and sample their delicious offerings. A scenic river valley of rolling hills, quaint farms, and mountain vistas, this area is also the heart of Cape Dutch culture. You'll see fine examples of traditional architecture while learning about the history of early settlement here. Lunch will be on your own in Stellenbosch.
Tonight, share your impressions of South Africa with your fellow travelers during a Farewell Dinner at a local restaurant.
hiddenDay 20Return to U.S.
After breakfast, you'll have the morning at leisure to do a little more exploring in Cape Town, or to simply relax before you begin your journey back to the U.S. this afternoon. Or begin your optional extension in Victoria Falls, Zimbabwe.


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text: I was just there couple weeks ago hunting the 3rd rifle elk season. We were in elk almost everyday. We saw huge herds of 100 plus animals. We hunted off our quads, but there is lots of hiking trails if you want to stay off the quad trails. Try bear gulch, there is always lots of elk in there and its accessible by hiking only.


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Ultraviolet light proven effective for restoration and continuous maintenance of coils
As air-conditioning equipment ages, its ability to maintain adequate space temperatures and humidity levels declines.1,2,3 Often, the culprit is reduced coil heat-transfer effectiveness, or the ability of air-handling-unit (AHU) cooling coils to remove heat from air. Evidence shows this drop in performance can occur within five years of startup. Typically, reduced heat-transfer effectiveness results from the buildup of contaminants on coil surfaces. This accumulation of organic material ...
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The Rolling Stones have booked their first ever show in Israel.
The rockers recently axed a tour of Australia and New Zealand following the death of Sir Mick Jagger's fashion designer girlfriend L'Wren Scott.
The band have now bounced back by adding a number of new shows to the upcoming European tour including dates in Norway and Portugal in May, Switzerland and Denmark in June, as well as a huge gig in Tel Aviv, Israel on June 4.
The promoter behind the Israel show, Shuki Weiss, confirmed the news on Tuesday and hailed The Rolling Stones visit as a huge step forward.
Weiss tells the Times of Israel, "This is a historic moment. It's a huge honor to bring the Rolling Stones to Israel, an honor for the country, the citizens, and mostly for the fans who have waited for this moment… This is emotional, even for me, and I've been doing this a long time."


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Worked With:
Warwick Davis
Birth Name: Warwick Davis
Born: 02/03/1970
Birth Place: Surrey, England, GB
Born: 02/03/1970
Birth Place: Surrey, England, GB
Born Feb. 3, 1970 in Surrey, England, Warwick Ashley Davis suffered from a rare genetic condition called spondyloepiphyseal dysplasia congenital (SED) which caused his dwarfism. He was attending the City of London Freemen's School when his grandmother heard an advertisement on the radio calling for people under four feet tall to appear in the final installment of the "Star Wars" trilogy at that time, "Return of the Jedi" (1983). A huge fan of the franchise, the two-feet, 11-inch-tall 11-year-old successfully auditioned, earning an extra role as an unnamed Ewok. When R2-D2 actor Kenny Baker, who was originally slotted to play Wicket, the lead Ewok, fell ill, Davis was given the part by George... Continue Bio >>
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|Carlos Diaz in tragic accident in Mexico
|Written by Brenda Johnson Brandt
|Wednesday, 15 May 2013 12:46
Ten days after saying goodbye to his family in Holyoke, Carlos Diaz, 48, was in a tragic accident in Mexico and he died May 7.
When he flew out of DIA in Denver April 25, it was Diaz's plan to go to Mexico to work for a time and then to hopefully return to his family in Holyoke.
He was riding with a friend in an Expedition Sunday, May 5, when they were involved in a tragic accident. His daughter Yazmin Diaz of Holyoke said the accident happened around 5:30-6 p.m., but they're not sure what occurred. The Expedition apparently had a flat tire while they were driving on the highway, and Yazmin said the vehicle might have rolled as a result.
Carlos was taken to a hospital in Chihuahua in critical condition. The driver in the accident was also critically injured and remains hospitalized.
By Tuesday, Yazmin said, doctors had determined that her dad had no brain activity. At that point, they decided to remove him from life support.
While it wasn't Yazmin's desire at first, all of the family surrounding Carlos wanted to have his organs donated. Yazmin said she eventually agreed. As a result, she was told, 46 people will benefit from her dad's organs, tissue, etc.
A funeral service was held in the Diaz family's hometown of Ignacio-Zargoza, Mexico on May 9, and a memorial was held at the same time at First Baptist Church of Holyoke, conducted by Ismael Lopez.
Yazmin said her two preschool-aged children, Brendan and Sujey, will dearly miss their grandpa, as they had a very close relationship. Yazmin describes her dad as a very pleasant man to all he met and a very, very hard worker.
In addition to Yazmin and her husband Salvador Bustillos and their children, other Holyoke family members include Carlos' wife Margarita, who works at Holyoke Marketplace; and his son, Carlos Jr., who is a junior at HHS and has qualified to compete in the state track meet this weekend.
Memorial account set up at Holyoke Credit Union
An account to help the Diaz family has been set up at Holyoke Community Federal Credit Union in the name of Carlos Diaz Erives.
Staff at Holyoke JR/SR High organized a benefit dinner for the family Tuesday night, May 14 as well. The plan was for 100 percent of the proceeds to go to the Diaz family. It was the staff's desire to help Carlos Jr.'s family be able to attend the state track meet to celebrate his success and also to help with expenses they have now and down the road.
Holyoke Enterprise May 16, 2013


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Pin-straight, perfect hair looks lovely for so many occasions – work or play, casual or formal. Flat iron heat tools for hair can quickly bring your hair from wild or wavy to perfectly straight, shiny, and beautiful. Flat irons can be just as easy to use as curling irons, making them a popular hair tool for anyone with medium length or long hair. The results of using these tools on your hair can last all day, giving you freedom from frizzies, fly-aways, and that dull look hair tends to get from time to time. A top quality flat iron like those in the CHI Air Flat Irons collection may produce a large amount of negative electricity, resulting in a smooth and silky hairstyle.
The CHI Air Flat Irons lineup relies on Tourmaline Ceramic Technology to provide even heat distribution and expert hair smoothing technique. The irons in the CHI Air Flat Irons collection come in a variety of sizes to achieve all looks for all hair lengths. Different colored flat irons like pink and red give you a fun choice in your hair styling tools. Choose a product from the CHI Air Flat Irons line to give your hair a gorgeous, straight look this season and all throughout the year.
Most flat irons typically come in slim 1/2-inch, classic 1-inch, or wide 1.5-inch styles to attend to a variety of hair lengths and desired styles. Some flat iron tools can even be used to style hair while it is still damp. Mini stylers and compact, travel flat irons round out the offerings so you are never left without the tools you need to keep your hair looking great anytime, anywhere. The CHI Air Flat Irons collection contains a wide variety of flat irons in different colors, sizes, and styles to suit all hair lengths and needs.
The CHI Air Expert Classic 1 inch Tourmaline Ceramic Flat Iron - Pure Pink expertly reduces static electricity, leaving hair soft, smooth, and silky. The pretty pink color makes this must-have flat iron a fun hair accessory to use all the time, creating shine, reducing frizz, and giving your hair a lovely straight look whenever you want it. This iron effectively straightens and smoothes your hair, but you can also use it to add a more versatile look – curling, waving, spiraling, and flipping various strands or the ends for your desired style.
The CHI Air Expert Classic 1 inch Tourmaline Ceramic Flat Iron - Fire Red, like its pink counterpart, is a slim Tourmaline ceramic flat iron that adds negative ions to reduce static and diminish frizz. The vibrant red color of the styler makes it especially enticing to add to your collection while the shiny and silky results are some you can't quite live without. Reviews show this classic flat iron is gentle on your hair, making it great for as-needed or everyday use.
The CHI Air Expert Classic 1 inch Tourmaline Ceramic Flat Iron - Onyx Black offers the same benefits as the pink and red versions above, giving you a choice in a classic black flat iron as well. This flat iron warms up fast and delivers results that last all day.


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Related Artists
The Miracles
Birth Place: Detroit, Michigan, United States
Years Active: 1955–1983, 1993–present
Genres: R&B, Soul
Years Active: 1955–1983, 1993–present
Genres: R&B, Soul
The Miracles (known from 1965 to 1972 as Smokey Robinson & The Miracles) are an American rhythm and blues group from Detroit, Michigan, notable as the first successful group act for Berry Gordy's Motown Record Corporation. Their 1960 single "Shop Around" was Motown's first million-selling hit record, and the group went on to become one of Motown's signature acts of the 1960s. During the same period, the Miracles' original lead singer and founding member Smokey Robinson became one of the most successful songwriters and record producers of all time. During their 19 year run on the American music charts, The Miracles charted over 50 hits and recorded in the genres of doo-wop, soul, disco, and ... Continue Bio >>
Listen to The MiraclesAll Albums >>
and similar artists - Play Now
and similar artists - Play Now
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Bodega Sans
Released in 1990, Bodega Sans adopts numerous ideas from the high period of Art Deco, providing designs that are as fresh as they are nostalgic. This geometric series, with its later seriffed companion, is unique in offering angular and nostalgically rounded forms of each weight, which designer Greg Thompson has called Regular and Oldstyle, as well as Regular Small Caps, offering unique typographic opportunities; FB 1990–92
Among other uses, Bodega Sans is recommended for Newspaper, Magazine, Book, Web and Corporate use.
Related People· Greg Thompson


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295.3J sec. 1 - McBaine Moot Court Competition (Spring 2014)
Instructor: William H. D. Fernholz (view instructor's teaching evaluations | profile)
View all teaching evaluations for this course
Units: 2
Class Number (formerly Course Control Number) (Non-1Ls): 50036
The James Patterson McBaine Honors Competition is Boalt Hall's venerable moot court competition and is open to all second- and third-year J.D. students. Cases chosen for the competition involve cutting-edge issues of great public importance. The Competition does not have a regular classroom component. Typically, the Board of Advocates arranges one or two non-mandatory lunchtime meetings to discuss the case and the relevant law. The student directors of the McBaine Competition and William Fernholz, the faculty director of appellate programs, will be available for individual meetings throughout the competition.
Judges, Boalt faculty, and appellate practitioners will evaluate the briefs and initial rounds of oral arguments. The preliminary, quarter, and semi-final rounds will take place on evenings in March and April. (Competitors with evening childcare responsibilities or other serious evening conflicts should contact William Fernholz with limitations; we will try to accommodate these conflicts.) Students will submit their briefs in late February or early March.
Students will advance to the quarter-finals, semi-finals and final based on their performance on the brief and in the preliminary rounds of oral argument, with the brief counting for at least half of the available points in the earlier rounds. Prizes are awarded for the best briefs and best oral argument.
Competitors receive 2 units for satisfactory participation in the competition. An introductory meeting will be held late in the Fall semester.
This course satisfies the Skills Requirement.
Exam Notes: T
This is a credit only course
Course Category: Simulation Courses
If you are the instructor or their FSU, you may add a file like a syllabus or a first assignment to this page.
No reader.
Instructor has indicated that no books will be assigned.


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l'âme soeur (lam sur) noun, feminine
: soul mate
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Audio file and example sentence: Listen to my daughter (then me, with a bit of difficulty...) pronounce these French words:
Pourquoi cette difficulté de trouver la véritable "âme sœur"? Peut être parce qu'il n'y en a qu'une sur Terre. Why is it so difficult to find one's true "soulmate"? Perhaps because there is only one of them on earth.
A Day in a French Life...
by Kristin Espinasse
My brother-in-law Jacques is in love. Love, love, love.
With Mariem.
They met two months ago in Avignon. Amour, amour, amour.
I met Mariem last week and she is everything I could wish for my beautiful beau-frère.* Mariem is lovely from the inside out. She glows.
And so does Jacques: il rayonne.* It must be love. Heaven knows.
We all deserve love, every one of us, but Jacques and Mariem especially so. Ils le méritent.* If I could I would compose a list of one hundred reasons why Jacques and Mariem especially deserve each other's love, but the list might turn into an epic novel. Besides, if I know you, you don't need convincing: you simply trust in the healing power of love....
Love covers a multitude of sins.
I do have a picture of our lovely Mariem... but because I have not asked permission to post it, and because Mariem is one to shun the spotlight, we'll have to settle on this one instead (thanks go to one of our puppies, "Sugar," who graciously offered to be a stand-in for the lovely Mariem):
To Jacques and Mariem: I think and I feel, I hope and I appeal that this love will endure à jamais*: forever.
Especially, I pray that it will. I know others do, too (Maman Jules, for example. Happy September 23rd birthday, Mom! I love you! And I will call you soon...).
One question I love to ask couples is: What is your secret to staying together?
Would you please, dear reader, share yours? Would you tell us your secrets to a love that lasts? I will share them with my brother-in-law and his lovely bride-to-be. Tee-hee!
...and I promise to read your comments, each and every one, to my own husband, just as soon as he wakes up tomorrow morning... to 15 years of marriage with me. God bless him!
So please share with us now your own relationship wisdom. What works for you? Whether you have been together 5 months or 50 years, please tell us your tips for a long-lasting love or simply give us one word that represents relationship endurance.
Click here to see reader-submitted tips or to share your own. Mille mercis!
~~~~~~~~~~French Vocabulary~~~~~~~~~~
le beau-frère (m) = brother-in-law; il rayonne = he is radiant, he shines; ils le méritent = they deserve it; à jamais = forever
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P.S.: I will be joining two authors at the American Library in Paris, to talk about "fish-out-of-water experiences" living in la belle France. If, by chance, you are in Paris on Oct. 7th, we would love to see you at this meet-up.


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Just as the first light of the first day of 2011 cracked in the East horizon today, January 1, 2011, the Washington Post reported how the U.S. efforts to convince the Pakistani Chief of The Armed Forces, General Ashfaq Kayani, to unleash his army against the Taliban have stalled. And that report – if someone reads between the lines- makes the U.S. effort in Afghanistan look more and more like a gravely ill misadventure that is sliding into another grave in the Graveyard of the Empires! History just will repeat itself, and Afghanistan would certainly become the second U.S. Vietnam for a superpower that the late Chinese leader Mao Tse-tung used to call "a paper tiger."
But as an American newspaper, the Washington Post cannot find anything wrong with the U.S. policy toward Pakistan, because that will offend and upset its readers and advertisers. The focus of the… Read more


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Mission Statement: Park University provides access to a quality higher education experience that prepares a diverse community of learners to think critically, communicate effectively, demonstrate a global perspective and engage in lifelong learning and service to others.Vision Statement: Park University, a pioneering institution of higher learning since 1875, will provide leadership in quality, innovative education for a diversity of learners who will excel in their professional and personal service to the global community.CourseMG 268 Office AdministrationSemesterS1DD 2012 DCXFacultyWade-Hairston, Tina A.TitleProfessor of Management Studies/Adjunct FacultyDegrees/CertificatesPh.D. Applied Management & Decision Sciences: Walden UniversityM.S. Administration: Central Michigan UniversityB.S. Computer Information Systems: Devry UniversityOffice LocationDSCC BLDG 11, Section 9Office HoursBy Appointment OnlyDaytime Phone614.557.1145Other email@example.com DatesS1DD 2012 DCXClass DaysU------Class Time12:00 - 5:00 PMPrerequisitesNoneCredit Hours3Textbook:
Title: Administrative Office Management
Author: Odgers
Edition: 13th
Publisher: THOMSON South Western
MG 268 Office Administration
Wade-Hairston, Tina A.
Professor of Management Studies/Adjunct Faculty
Ph.D. Applied Management & Decision Sciences: Walden UniversityM.S. Administration: Central Michigan UniversityB.S. Computer Information Systems: Devry University
Office Location
DSCC BLDG 11, Section 9
Office Hours
By Appointment Only
Daytime Phone
Other Phone
Semester Dates
Class Days
Class Time
Credit Hours
Textbooks can be purchased through the MBS bookstore
Textbooks can be purchased through the Parkville Bookstore
Additional Resources:
McAfee Memorial Library - Online information, links, electronic databases and the Online catalog. Contact the library for further assistance via email or at 800-270-4347.Career Counseling - The Career Development Center (CDC) provides services for all stages of career development. The mission of the CDC is to provide the career planning tools to ensure a lifetime of career success.Park Helpdesk - If you have forgotten your OPEN ID or Password, or need assistance with your PirateMail account, please email firstname.lastname@example.com or call 800-927-3024Resources for Current Students - A great place to look for all kinds of information http://www.park.edu/Current/.
Educational Philosophy: The faculty's
educational philosophy is one of empowerment. Students are challenged to strive
for great achievements both personally and professionally. The learning
environment will be used as a platform for debate, an exchange of ideas, and a
vehicle to shape developing perspectives. Academic excellence will be
encouraged and supported through readings, group activities, and examinations.
Learning Outcomes: Core Learning Outcomes
The assessment device will be a final paper in which the Core Learning Outcomes are addressed by using the internet to research the most recently published research about issues, products and services that relate to the textbook material.
Link to Class RubricClass Assessment: A variety of methods
will be used to assess the learning that occurs in this course. Students will
be assessed on their reflection Journals, mid-term analysis, research paper,
and performance on the final examination.
Reflection Journals - Five @ 50 points each = 250
Mid-term Analysis – One @ 250 points each = 250 points
Research Paper = 200 points
Final Exam = 300 points
Course grade distribution is as follows:
A = 90% - 100% (900-1000 points)
B = 80% - 89% (800-899 points)
C = 70% – 79% (700-799 points)
D = 60% - 69% (600-699 points)
F = Less than 60% (599 points and
The course grade for students will be based on the overall average of homework and tests taken during the course in accordance with the weighting of the various requirements as stated in the syllabus.
All final exams in all School of Business courses will be comprehensive and will be closed book and closed notes. They will constitute 30% of the total course grade and will not be a take-home exam. They will be completed during the test week in the period designated by the registrar or by the Proctor in the case online courses. If calculators are allowed, they will not be multifunctional electronic devices that include features such as: phones, cameras, instant messaging, pagers, and so forth. Electronic Computers will not be allowed on final exams unless an exception is made by the Dean of the School of Business.
Classroom Rules of Conduct:
1. Students
are expected to show respect to fellow students and to decisions made by the
2. Students must be prepared for class. This includes having
course materials such as the required text for class.
3. Students that do not have the required course materials will
not be afforded extensions for in-class and/or home assignments.
4. Students must notify the instructor in advance of absences
via email. phone, etc.
Course Topic/Dates/Assignments:
Class Schedule
Course Activities
Homework Assignments
One – January 17
1, 2
Chapters 1, 2
Two – January 24
3, 4, 5
Chapters 3, 4, 5
Journal # 1 due
Three – January 31
Chapters 6, 7
Journal # 2 due
Four – February 7
8, 9
Chapters 8, 9
Journal # 3 due
Five – February 14
10, 11
Chapters 10, 11
Mid-term analysis Due
Six – February 21
12, 13
Chapters 12, 13
Journal # 4 due
Seven – February 28
14, 15
Chapters 14, 15
Journal # 5 due
Eight – March 6
Research Paper and Final Exam due
Academic Honesty:Academic integrity is the foundation of the academic community. Because each student has the primary responsibility for being academically honest, students are advised to read and understand all sections of this policy relating to standards of conduct and academic life. Park University students and faculty members are encouraged to take advantage of the University resources available for learning about academic honesty (www.park.edu/current or http://www.park.edu/faculty/).from Park University 2011-2012 Undergraduate Catalog Page 93
Plagiarism:Plagiarism involves the use of quotations without quotation marks, the use of quotations without indication of the source, the use of another's idea without acknowledging the source, the submission of a paper, laboratory report, project, or class assignment (any portion of such) prepared by another person, or incorrect paraphrasing. from Park University 2011-2012 Undergraduate Catalog Page 93
Attendance Policy:Instructors are required to maintain attendance records and to report absences via the online attendance reporting system.
Park University 2011-2012 Undergraduate Catalog Page 96
Disability Guidelines:Park University is committed to meeting the needs of all students that meet the criteria for special assistance. These guidelines are designed to supply directions to students concerning the information necessary to accomplish this goal. It is Park University's policy to comply fully with federal and state law, including Section 504 of the Rehabilitation Act of 1973 and the Americans with Disabilities Act of 1990, regarding students with disabilities. In the case of any inconsistency between these guidelines and federal and/or state law, the provisions of the law will apply. Additional information concerning Park University's policies and procedures related to disability can be found on the Park University web page: http://www.park.edu/disability .
Additional Information:
Attachments:Assignment OverviewBibliography:
Last Updated:1/12/2012 3:33:55 PM


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St. Louis County (KSDK) - The Ladue Board of Education has accepted a request for personal leave by Dr. Marsha Chappelow, Superintendent of Ladue Schools.
Dr. Chappelow's leave will begin December 1, 2012 and last through her retirement on June 30, 2013.
She will be able to still provide consultative and transition service through the superintendent search and will be available to answer questions pertaining to district operations.
"The Board is moving forward and has selected a search firm to assist us in our superintendent selection process and we, with Dr. Chappelow's leadership, have made steady progress on our Board goals for this year. We want to assure our school community that the District will continue to provide the highest level of education, planning, and service during this transition period," said Board President Jayne Langsam.
Dr. Chappelow requested leave for undisclosed personal reasons.


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July 13, 2013 | 1064 views
Challenging themselves physically and mentally
Hewlett residents to compete in third Ironman Triathlon
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Cary Epstein and Jay Greenbaum, both Hewlett residents, along with Steven Gartenstein of Hewlett Harbor, are preparing to compete in their third Ironman Triathlon on July 28.
The trio will swim 2.4 miles, bike 112 miles and run 26.2 miles at the Lake Placid-based event. "The Ironman Triathlon is named 'the grand daddy of all endurance races,' it is hands down the toughest one-day endurance event in the world," Epstein said. "It tests both your physical and mental limitations; it's not for the faint of heart."
Having competed in his first triathlon 13 years ago, Cary Epstein couldn't have imagined he would still be participating. "The last 13 years have been an incredible journey for me in the sport of triathlons," he said. "I never thought I would do another triathlon after the first race and here I am about to go into my third Ironman."
Epstein, a former health education teacher and head coach of the girl's varsity swimming and diving team as well as assistant coach for the boy's swimming and diving team at Hewlett High School, is also a member of the 40-person Ironman Triathlon Team. "I filled out an application just for fun; I didn't think I had a shot in hell," he said. Two months went by and I received a phone call from them saying I had been chosen from thousands of applicants across the U.S. I was in shock and I couldn't be more excited to be apart of such an incredible group and give back to the Ironman Communities who host our races."
Greenbaum participated in an Ironman Triathlon last August in Manhattan and again in March in Melbourne, Australia. He's competed in 25 triathlons over the past eight years. "I enjoy the multi-sport discipline," he said. "Rather than simply focusing on one sport, having to become proficient in all three components: swimming, biking and running, breaks up the monotony to a degree."
Gartenstein has competed in more than 30 races. "My first triathlon was an Olympic distance triathlon in Montauk," he said. "The weather was terrible and the water was very cold but I had a great time and I was hooked on racing and the competition."
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Doppler Shift
Application ID: 1376
We have all noticed the Doppler effect when an ambulance passes by with its sirens blaring. The siren's pitch suddenly drops the moment the ambulance starts moving away from you.
Another effect you can notice is how the siren's sound is very loud when nearby, but as soon as the ambulance has passed the sound becomes much quieter.
This model simulates the acoustic Doppler effect by placing a stationary sound source in a constant fluid flow.
The frequency shift is visualized, and the acoustic intensity, as measured by an observer moving with the flow, is calculated.


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Morning game after night game!
The Royals and Rangers are playing a B-game this morning at 9:30 Arizona time (11:30 K.C.). If you're in Surprise, check it out. Several regulars are in the lineup:
(Podsednik – LF), (Betemit – SS), (DeJesus – CF), (Butler – 1B), (Fields – 3B), (Aviles – 2B), (Pena – C), (Falu – RF), (Getz – DH), (Hernandez – P)


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With a complete project renovation to show off, the new ownership and management of Rifle Creek Apartments, formerly Rimrock Apartments, will hold an open house from 2-5 p.m. Wednesday, Dec. 19, at 3025 Coal Mine Ave. in Rifle.
Renovations began in July, shortly after Rifle Creek Apartments LLC took ownership of the property. All 11 buildings received new windows and paint. The entire property received a landscaping overhaul that included new paved parking lots, sod and landscaping materials, plus a dog park. Unfinished building foundations from the original construction that were unsightly and unsafe were removed and landscaped to create additional open space. Facilities Contracting oversaw the renovations.
Rifle Creek Apartments includes 94 studio, one-, two- and three-bedroom homes. The homes are receiving internal upgrades as well, including new carpet and appliances.
Leases range from $550 to $750 per month. Residents who sign a lease in December will not pay rent until January. Corporate leases are available. For more information, visit www.riflecreekapts. .com.
WingNutz Bar and Grill will once again provide ham dinner packages for families in the Rifle area on Christmas Eve.
If you or someone you know would enjoy this dinner, call 625-9300. Sign up by Dec. 19, as dinners are limited and available on a first-call, first-serve basis. Delivery will be available, if needed, in the Rifle area.
Our daughter, Mackenzie, recently participated in the 2012 Miss American Princess pageant at Disney World in Florida. She raised $1,300 in donations from area businesses and residents in order to attend.
We want to thank these sponsors: TWI Oilfield Fabrication, Cheryl Chandler Real Estate, Rifle Truck and Trailer, Pediatric Partners, Valley View Hospital, Miller's Dry Goods, Mountain View Tree Farm, Hogback Pizza, Swallow Oil, Hy-way Feed, Dorothy Day at the Bookcliff Gas Station, Western Slope Jeep Chrysler Dodge, Enstrom's, Mr. Dillon, Todd and Vivian, Paul Fasold, Grandpa Dave, Grandma, Grammie and Grandpa Jamie, Mommy, Daddy and Mataya, JoJo and Steve.
Mackenzie, Tina and Darin Holtz
On behalf of Wamsley Elementary School, I would like to thank All Kids Dental for their presentation on good dental hygiene to our students on Monday, Dec. 3. We greatly appreciate their years of continued commitment to the health and wellness of our Wamsley kids, not to mention the donation of tooth brush kits for all the children.
Thank you, All Kids Dental.
Desha Bierbaum, principal, Rifle
We have watched you play the sports you love since you were little boys. It has been such a blessing to watch your skills improve each year, but even more to watch you grow into such special young men.
The last two years have been a wonderful ride, you should be proud. We are.
Thank you team, thank you coaches, thank you parents and fans for your sacrifices. God bless you all. Go Bears!
Carleton and Heather Hoffmeister
The winners of the downtown window display contest on Saturday, Dec. 1, sponsored by the Rifle Downtown Development Authority and visitors information fund, were picked through a people's choice vote:
First place: $100, Blown Away Day Spa; second place: $75, TrendZ; third place: $50, New Image Boutique.


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Track-State-World Record: 1:13 Sabertooth (7), May 22, 2005Stakes Record: 1:14 3/5, Gadget Queen (2009); 1:14.61, E Z Kitty (2012)Most Wins*Horse: 1, 18 tiedJockey: 3, Ricky Frazier (A Classic Life, 2006; She's All Silk, 2007; Gadget Queen, 2009)Trainer: 1, 18 tiedOwner: 2, Seawind Stables (Banchory Baron, 1996; Russellthemussell, 1997) Largest Winning Margin: 6 lengths, Ema Bovary (2003Smallest Winning Margin: nose, Always a Dixie (2002), A Classic Life (2006)Winner bred: Washington 9, Kentucky 4, British Columbia 2, Florida 2, Chile Betting favorite: 2-for-18 (11%), Odds-on: 2-for-7 (14%)Purse: 1st—$27,500-2nd—$10,000-3rd—$7,500-4th—$3,750-5th—$1,250*Race was for 3-year-old colts and geldings prior to 2000
Owners Hometowns1—Tabled With Gold (12) Patricia Zacker, La Quinta, CA2—Blueberry Smoothie (6) Northwest Farms LLC (Jerre Paxton, Yakima, WA); C & M Racing (Corey Kinder, Yakima, WA; Mihai Tiru, Bellevue, WA)3—Finding More (10) Blue Ribbon Racing #11 (Fred & Debbie Pabst, Buckley, WA)4—Goin to the Window (8) Northwest Farms LLC (Jerre Paxton, Yakima, WA) Debbie Paxton, Yakima, WA5—Madame Pele (9-2) Kama'aina Thoroughbreds (Zach & Beth Correa, Enumclaw, WA)6—Stopshoppingdebbie (1-2) Northwest Farms LLC (Jerre Paxton, Yakima, WA)
3 & Up Fillies & Mares @ Emerald DownsMay 11: $50,000 Hastings Handicap, 6 Furlongs, Stopshoppingdebbie ($3) 1:08.38June 8: $50,000 WA State Legislators Stakes, 6-½ FurlongsJuly 6: $50,000 Boeing Handicap, 1MileAug. 24: $65,000 Emerald Distaff, 1-1/8 MileSept. 7: $50,000 Pegasus Training Center Stakes (Washington-bred), 1-1/16 Mile
FAB 5 HANDICAPPERSJoe Withee (EmD1): Stopshoppingdebbie, Madame Pele, Goin to the Window (1-4; $40.30)Robert Geller (EmD2): Stopshoppingdebbie, Goin to the Window, Madame Pele (2-4; $55.50)Jacob Pollowitz (EmD3): Stopshoppingdebbie, Goin to the Window, Madame Pele (3-4; $42.70)Vic Cozzetti (Predictorsays.com): Stopshoppingdebbie, Blueberry Smoothie, Goin to the Window (2-4; $40.90)Nick Rousso (Daily Racing Form): Stopshoppingdebbie, Madame Pele, Finding More (2-4; $32.10)Consensus: Stopshoppingdebbie (25), Goin to the Window (6), Madame Pele (6) (2-4; $37.20)
NOTES: Stopshoppingdebbie would become the first horse since Marva Jean in 2005 to sweep the Hastings Handicap & Washington State Legislators Stakes in the same year…Exclusive Diva—trained by Tom Wenzel and owned by Northwest Farms—denied E Z Kitty a sweep of last year's Hastings/Washington State Legislators with a 1 ¼-length win in 1:14.88. E Z Kitty finished second at 0.90-to-1 and became the 10th straight favorite to lose in the Washington State Legislators, a streak that began when Northwest Farms' Lasting Code finished third at 0.70-to-1 in 2004. Subsequent favorites to lose: Sariano, 4th at 1.70-to1 in 2005; Sudden Departure, 6th at 3.40-to-1 in 2006; Kissntheboysgoodby, 3rd at 2.00-to-1 in 2007; Private Dreams, 5th at 0.60-to-1 in 2008; High Resolve, 2nd at 0.30-to-1 in 2009; Hallelujah Trail, 2nd at 1.40-to-1 in 2010; Cliff's Secret, 5th at 1.20-to-1 in 2011; Class Included, 2nd at 1.10-to-1 in 2012. . .Fleet Pacific in 2001 and Ema Bovary in 2003 are the only betting favorites to win this race. When Fleet Pacific prevailed by a half-length in 2001, finishing second at 22-to-1 was Taste the Passion, dam of Stopshoppingdebbie. . .Shampoo, dam of Blueberry Smoothie, finished third in the 2008 Washington State Legislators. . .With a victory, Stopshoppingdebbie would become only the 10th horse to win at least six stakes at Emerald Downs. . .With a victory¸ Stopshoppingdebbie would have the most wins by an undefeated horse in North America; she enters Sunday tied with Ide Be Cool, a 3-year-old gelding based at Delta Downs.


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ACHICA > Argent of London > Argent Of London Lattice Design Enamel Egg Pendant
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Argent Of London Lattice Design Enamel Egg Pendant
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Bill Crosier is a tinkerer. He's a specialty engine mechanic by trade, but he says most of what he does is tinkering.
Ask anybody in his community of motorsports enthusiasts and machine operators, and you may hear that he is a master of his craft.
That craft is much harder to do these days, after a fire destroyed and damaged thousands of dollars of special equipment and tools Crosier has collected over the years.
The small business he runs out of the garage at his South Missoula home doesn't bring in enough revenue to replace the damaged items, let alone the man hours it will take to restore what is not completely gone.
Reed Hunter is a good friend and frequent customer of Crosier's, and has been a major contributor to the efforts set forth by the community to help Crosier get back to work.
"Bill's been so good to everyone in the community for so long, that we thought we needed to try and organize and see if we could help him out because he's helped an awful lot of us for a long time."
Men young and old have been flocking to Crosier's property to help lend a hand, but it takes more than the average Joe to clean some of the damaged equipment.
"Some of that stuff is so particular," Hunter says, "you really have to know what your doin' or you can cause more damage than good."
People who aren't able to lend a hand to the cause have started a benefit fund through First Security Bank.
Right now the savings are modest. 3,000 dollars is not near enough to pay for the 50 some odd thousand dollars in damages that Crosier estimates.
Anybody who would like to offer help to his cause can take a donation to any First Security Bank location or by mailing Bill Crosier Benefit Fund, First Security Bank. You can also mail donations to P.O. Box 4506, Missoula MT. 59806