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The Runt-related transcription factor Runx2 is essential for bone development but is also implicated in progression of several cancers of breast, prostate and bone, where it activates cancer-related genes and promotes invasive properties. The transforming growth factor β (TGF-β) family member bone morphogenetic protein-3B (BMP-3B/GDF10) is regarded as a tumor growth inhibitor and a gene silenced in lung cancers; however the regulatory mechanisms leading to its silencing have not been identified.
Here we show that Runx2 is highly expressed in lung cancer cells and downregulates BMP-3B. This inverse relationship between Runx2 and BMP-3B expression is further supported by increased expression of BMP-3B in mesenchymal cells from Runx2 deficient mice. The ectopic expression of Runx2, but not DNA binding mutant Runx2, in normal lung fibroblast cells and lung cancer cells resulted in suppression of BMP-3B levels. The chromatin immunoprecipitation studies identified that the mechanism of Runx2-mediated suppression of BMP-3B is due to the recruitment of Runx2 and histone H3K9-specific methyltransferase Suv39h1 to BMP-3B proximal promoter and a concomitant increase in histone methylation (H3K9) status. The knockdown of Runx2 in H1299 cells resulted in decreased histone H3K9 methylation on BMP-3B promoter and increased BMP-3B expression levels. Furthermore, co-immunoprecipitation studies showed a direct interaction of Runx2 and Suv39h1 proteins. Phenotypically, Runx2 overexpression in H1299 cells increased wound healing response to TGFβ treatment.
Our studies identified BMP-3B as a new Runx2 target gene and revealed a novel function of Runx2 in silencing of BMP-3B in lung cancers. Our results suggest that Runx2 is a potential therapeutic target to block tumor suppressor gene silencing in lung cancer cells.
Keywords:Lung cancer; Runx2; BMP-3B; Gene silencing
Lung cancer is the leading cause of cancer mortality and accounts for 30% of all deaths from cancer [1]. Silencing of tumor suppressor genes by aberrant promoter hypermethylation is a key event in lung cancer initiation and progression. During gene silencing, the chromatin structure is altered by acetylation, phosphorylation and methylation of histone tails [2]. These alterations in chromatin structure affect normal cell functions and are a crucial trigger for neoplastic development and progression [3]. However, current understanding of regulatory mechanisms of silencing of tumor suppressors is limited. In this study we identified a mechanism by which Runx2 transcription factor contribute to epigenetic silencing of a tumor growth inhibitor BMP-3B in lung cancer cells.
Runx transcription factors (Runx1, Runx2 and Runx3) are critical regulators of organogenesis and cell differentiation regulatory pathways, and mutations in these genes are associated with several cancers. Runx2, an essential bone cell differentiation factor [4,5] is recently implicated in mammary, prostate and osteosarcoma progression [6-8]. In cancer cells, Runx2 activates cancer-related genes, promotes cells invasive properties [6,8-10], cooperates with oncogenes (e.g., c-myc in T-cell lymphoma development), and suppresses apoptotic and growth arrest pathways [11,12]. Runx2 is also a major target gene of TGFβ /BMP signaling pathway and the interaction between Runx2 and Smads results in regulation of downstream target genes in osteoblasts [13], chondrocytes [14] and cancer cells [8].
BMP-3B, a TGFβ family member and closely related to BMP-3, is highly expressed in lung [15-17], brain and bone tissues, and induces bone formation [18,19]. Ectopic BMP-3B expression promotes osteoblast differentiation and augments the bone formation induced by bone morphogenetic protein-2 (BMP-2) in rats [20]. Importantly, the expression of BMP-3B is downregulated in lung cancer patient samples and cancer cells lines compared to normal lung cells [21-23]. Multiple mechanisms have been proposed for the downregulation of BMP-3B levels which include methylation of gene promoter and repression by transcription factors [21] however, the transcriptional repressor proteins of BMP-3B are unknown.
We show that BMP-3B is a novel Runx2 target gene and find an inverse relationship between Runx2 and BMP-3B expression levels in normal lung fibroblast and lung cancer cells. Our studies with Runx2 overexpression or knockdown in lung cancer cells indicate that Runx2-mediated downregulation of BMP-3B is via increasing histone H3K9 methylation status of the proximal promoter by interacting with methyltransrefase Suv39h1.
Calvarial mesenchymal cells of Runx2-deficient mice have higher expression levels of BMP-3B
To identify novel Runx2 target genes, we performed cDNA expression analysis on total RNA isolated from calvarial mesenchymal cells of wild type and functional deficient Runx2 mice [5]. In addition to the downregulation of known Runx2 target genes (e.g., matrix metalloproteinases) in a osteogenesis-related cDNA array [24], we found that the expression levels of BMP-3B gene was induced in Runx2 deficient cells compared to wild type cells (Figure 1a). The induction of BMP-3B expression in Runx2 deficient calvarial mesenchymal cells was validated by qRT-PCR analysis (Figure 1b). To further confirm Runx2-mediated downregulation of BMP-3B levels, we re-expressed Runx2 via adenoviral delivery in Runx2 deficient primary calvarial cells and measured BMP-3B levels by qRT-PCR analysis (Figure 1c). Our results show a dose-dependent repression of BMP-3B mRNA levels by Runx2 in primary osteoblastic cells. These results suggested that BMP-3B is a novel Runx2 responsive gene.
Figure 1. Runx2 deficient mice have higher levels of BMP-3B. (a) Total RNA from mesenchymal cells of calvarial tissue from wild type and Runx2-/- (17.5 dpc) was hybridized with osteogenesis-related cDNA array. Signal of BMP-3B on blots is indicated by an arrow. Control genes (GAPDH, cyclophilin A) are shown in lower panel. (b) BMP-3B mRNA levels (normalized to GAPDH) in calvarial mesenchymal cells from wild type (WT) and Runx2-/- animals as detected by qRT-PCR analysis. (c) Primary calvarial cells from Runx2 deficient animals were transduced with empty vector (EV) or Runx2 expressing adenovirus with increasing multiplicity of infection (MOI 10 and 20) for 48 hr. Total RNA isolated from the infected cells was utilized to examine BMP-3B mRNA as detected by qRT-PCR analysis. (d) Runx2 mRNA expression levels were examined by qRT-PCR and the gene expression levels were normalized to 28S internal control. The gene expression levels were calibrated to IMR-90 cells. (e) BMP-3B mRNA expression levels were examined by qRT-PCR and the gene expression levels were normalized to 28S internal control. The BMP-3B gene expression levels were calibrated to Runx2 levels in IMR-90 cells. (f) Runx2 protein levels were examined in nuclear extracts of normal lung fibroblast cells (IMR-90 and WI-38) and lung cancer cell lines (H720 and H1299) by immunoblotting with Runx2 monoclonal antibody. Runx2 expression levels were normalized to internal control LaminA/C protein. (g) Runx2 intracellular localization was determined by immunofluorescence of endogenous Runx2 protein in WI-38 and H1299 cells. The punctate (Alexa 488, green) signal shows Runx2 staining while nuclei are revealed by 4, 6-diamidino-2-phenylindole (DAPI, blue) staining.
An inverse relationship between Runx2 and BMP-3B expression levels in lung cancer cells
A tumor growth inhibitory function was proposed for BMP-3B in lung cancers and BMP-3B is downregulated in most of the lung cancers [22,23,25]. In context of Runx2-mediated BMP-3B suppression in mesenchymal cells and to understand the upstream regulatory mechanisms of BMP-3B silencing in lung cancers, we hypothesized that Runx2 downregulates BMP-3B expression in lung cancer. To understand the role of Runx2 in BMP-3B transcriptional regulation in lung cancer cells, we first examined Runx2 and BMP-3B mRNA levels in normal lung fibroblasts of mesenchymal origin (WI-38 and IMR-90), atypical carcinoid (H720) and metastatic non-small cell lung carcinoma (H1299) cells by qRT-PCR analysis. Our results showed that Runx2 expression is increased in metastatic lung cancer cells (H1299) compared to normal lung fibroblast cells. In contrast to the Runx2 expression levels, BMP-3B mRNA was detectable but lower in lung cancer cells compared to normal lung fibroblast cells (Figure 1d and e). The Western blot analysis for Runx2 protein levels further validated increased Runx2 levels in lung cancer cells compared to normal lung fibroblast cells (Figure 1f). A punctate nuclear staining of Runx2 was observed in WI-38 and H1299 cells as examined by immunofluorescence (Figure 1g). Taken together, these studies revealed that the inverse relationship between Runx2 and BMP-3B levels observed in calvarial mesenchymal cells also holds true for normal lung fibroblasts and lung cancer cells.
Runx2 overexpression suppresses BMP-3B in lung cancer cells
To investigate whether Runx2 suppresses BMP-3B levels in lung cancer cells similar to observed in primary calvarial cells, we stably overexpressed wild type Runx2 (WT) and Runx2 DNA binding domain mutant (DBD) in normal lung fibroblast cells (WI-38 and IMR-90) by lentiviral-mediated gene delivery. Expression levels of wild type and mutant Runx2 protein in these cell types were confirmed by qRT-PCR and western blot analysis (data not shown). Our results showed that stable expression of wild type Runx2 in normal lung cells resulted in more than 2-fold decrease in BMP-3B levels compared to empty vector control cells (Figure 2a and b). Ectopic expression of DBD mutant of Runx2 failed to downregulate BMP-3B levels in normal lung or lung cancer cells. These results suggested that the Runx2 DNA binding activity is required for BMP-3B regulation. In complementary studies, Runx2 knockdown resulted in increased BMP-3B levels in normal bronchial NL-20 cells (10- fold increase, Figure 2c right panel) and H1299 cells (2-fold increase, Figure 2d right panel) compared to empty vector controls as shown by qRT-PCR analysis. The decrease in Runx2 levels in Runx2 knockdown cells was confirmed by qRT-PCR and western blot analysis (Figure 2c and d). Collectively, these results indicate that Runx2 downregulates BMP-3B levels in normal lung fibroblast and lung cancer cells.
Figure 2. Runx2 suppresses BMP-3B in lung cancer cells. (a and b) Expression levels of BMP-3B in normal lung fibroblast cells (a: WI-38, b: IMR-90) transduced with wild type Runx2 (WT), functional deficient mutant (DBD: DNA binding domain mutant,) and empty vector (EV) control lentiviral particles are shown as detected by qRT-PCR analysis. (c and d) The Runx2 protein was suppressed in normal bronchial NL-20 epithelial cells and H1299 lung cancer cells by lentiviral vector-mediated RNAi. The suppression of Runx2 at mRNA and protein expression levels is shown for NL-20 cells (C top left and lower left panels). The mRNA levels of BMP-3B in Runx2 suppressed NL-20 and control cells are shown as examined by qRT-PCR (C top right panel). The suppression of Runx2 at mRNA and protein expression levels is shown for H1299 cells (D top left and lower left panels). The mRNA levels of BMP-3B in Runx2 suppressed H1299 and control cells as examined by qRT-PCR (D top right panel).
Runx2 recruitment on the BMP-3B gene promoter and interaction with Suv39h1 promotes BMP-3B silencing
To further investigate the mechanism of Runx2-mediated downregulation of the BMP-3B expression in lung cancer cells, we performed chromatin immunoprecipitation analysis in H1299 cells expressing either wild type Runx2 or shRunx2 (Figure 3). Our results showed 3-fold increased Runx2 binding on the BMP-3B proximal promoter (-1.0kb) in H1299-WT-Runx2 cells, that was abrogated in H1299-shRunx2 cells. We next examined the methylation status of the BMP-3B proximal promoter as methylation of lysine 9 of histone H3 (H3K9) allows the binding of heterochromatin protein- 1 (HP1) to silence gene expression [26,27]. Our results show increased (3-fold) H3K9 levels of proximal promoter region of BMP-3B in H1299-Runx2 cells compared to H1299-shRunx2 cells or antibody controls (Figure 3a). We next examined the recruitment of Suv39h1 protein, a histone H3 lysine 9 specific methyltransferase, on BMP-3B proximal promoter. A twofold increase in recruitment of Suv39h1 was observed in H1299-Runx2 cells compared to H1299-shRunx2 lung cancer cells (Figure 3a). These findings indicated the possibility of physical interaction of Runx2 and Suv39h1 proteins in lung cancer cells. We performed co-immunoprecipitaion assays with Runx2 and Suv39h1 antibodies and a direct interaction of Runx2 with Suv39h1 proteins was detected in H1229 cells (Figure 3b). Taken together, these results show that the recruitment of Runx2 and Suv39h1 on the BMP-3B proximal promoter sequences resulted in increased H3K9 methylation status and consequently downregulation of BMP-3B expression in lung cancer cells.
Figure 3. Runx2 recruitment on BMP-3B gene promoter and interaction with Suv39h1. (a) Lung cancer cells (H-1299) stably expressing wild type Runx2 (left panel) or shRNA-Runx2 (right panel) were subjected to chromatin immunoprecipitation assay with Runx2, Suv39h1, H3K9 or control IgG-G (goat) and IgG-R (rabbit) antibodies. Immunoprecipitated DNA samples were further amplified by real-time qPCR with primer pairs for proximal promoter (-1.0kb) of BMP-3B gene. A schematic diagram above indicates Runx binding motifs (solid bars) on BMP-3B promoter. The BMP-3B ChIP primer pairs are indicated by arrows in the schematic diagram. (b) Co-immunoprecipitation assay shows Runx2 interaction with Suv39h1 in H-1299 lung cancer cells. Whole cell extracts from lung cancer H-1299 cells were incubated with Runx2, Suv39h1 or control IgG antibodies. Western blots showing Runx2 and Suv39h1 proteins as detected by αRunx2 and αSuv39h1 antibodies.
Runx2 increases wound-healing response of lung cancer cells
To examine the phenotypic effects of Runx2 overexpression in lung cancer cells, we assessed proliferation and migration potential of H1299-Runx2 cells or H1299- empty vector cells. Increased Runx2 levels in H1299-Runx2 cells and a corresponding decrease in BMP-3B mRNA expression were confirmed by western blot and qRT-PCR analysis respectively (Figure 4a). A 40% decline in cell proliferation was observed in Runx2 overexpressing H1299 cells compared to empty vector control cells in absence or presence of TGFβ treatment as examined by cell growth assay (data not shown) and MTT assays (Figure 4b). However, in response to TGF-β treatment the Runx2 overexpression in H1299 cells resulted in a significant (p <0.05) increase in wound healing response compared to the empty vector control for 6-48h as shown by wound healing assay (Figure 4c). The H1299 EV or WT-Runx2 cells did not show any differences in KI-67 (cell proliferation marker) immunoreactivity around wound area (data not shown). These results suggest that Runx2 promotes migratory potential of lung cancer cells by modulating TGF-β/BMP-3B signaling axis.
Figure 4. Runx2 increases migration potential of lung cancer cells. (a) Left panel: Runx2 protein was overexpressed in H-1299 cells via lentiviral vector-mediated gene transfer and overexpression levels are shown by Western blot analysis. Right panel: The mRNA levels of BMP-3B in Runx2 overexpressed H1299 cells as examined by qRT-PCR (b) The Runx2 overexpressing H1299 cells or empty vector (control) were seeded in a 96 well cell culture dish in presence or absence of TGFβ (5ng/ml) and examined for changes in cell proliferation via cell proliferation MTT assay by measuring absorbance at 490 nm. (c) Cell migration in H-1299 cells expressing Runx2 or empty vector control was examined in a standard wound/scratch healing assay in response to TGF-β treatment. Representative images at 40X magnification and average (±standard deviation) wound healing response is shown at indicated time points. (* P ≤ 0.05).
Our studies identify BMP-3B as a Runx2 target gene and show that Runx2 promotes epigenetic silencing of BMP-3B in lung cancer cells by promoting histone H3K9 methylation status of the proximal regulatory regions. The Runx2 interaction with Suv39h1 methyltransferase and binding to the BMP-3B promoter results in downregulation of the BMP-3B expression levels. Furthermore, ectopic expression of Runx2 enhances the migration potential of lung cancer cells in response to the TGFβ signaling.
We find that mesenchymal cells from Runx2-deficient animals express high levels of BMP-3B compared to wild type cells. In contrast to high levels of BMP-3B, low baseline levels of BMP2 are reported in Runx2 deficient cells that can be up-regulated by ectopic expression of Runx2 [28]. Interestingly, a BMP2 orthologous signaling antagonizing function for BMP3/3B has been proposed during embryonic development of xenopus [29]. In addition to directly regulating expression levels of BMP family members as shown by these studies, Runx2-Smad complex has been shown to regulate expression of genes related to osteogenic [10,30,31] and cancer [32] properties in response to TGFβ/BMP signaling. The consequences of direct regulation of BMP-3B by Runx2 on downstream molecular events of TGFβ/BMP pathway still need to be determined. A recent report shows that the migration of lung cancer cells is associated with the upregulation of Runx2 and Snail expression in response to BMP-2 treatment [33]. Our results show that Runx2 downregulates BMP-3B and increases migration potential of lung cancer cells in response to TGFβ treatment. These studies suggest that cross-talk between Runx2 and TGFβ/BMP signaling is differential and could be context-dependent.
Our results showing higher gene and protein expression levels of Runx2 in lung cancer cells compared to normal lung fibroblast cells are consistent with previous reports of Runx2 expression in other epithelial cancers like breast and prostate cancers [8,10,31,34-36]. The Runx2 gene expression levels were similar in IMR-90 and WI-38 cells; however BMP-3B levels were dramatically reduced suggesting cell-type-specific differences. In addition, we find that the Runx2 overexpression in lung cancer cells results in a significant decline in cell proliferation but enhances wound healing response. In serum-deprived conditions used for the wound healing assay, we observed similar numbers of KI-67 positive cells close to (200-400μm) wound area in both EV and WT-Runx2 over-expressing cells. As we find KI-67 positive cells in both groups, therefore, we cannot completely rule out the possible contribution of cell proliferation in the observed wound healing phenotype. This phenotype is probably the combinatorial effect of Runx2 on BMP-3B suppression and activation of genes related to invasion and migration (e.g., MMPs), as Runx2 is known to promote migration and invasive potential of breast and prostate cancer cells [6,8,9,24,33,35,37-39]. The downstream molecular events of BMP-3B silencing in lung cancer progression are still not clear and might include phosphorylation of Smad proteins as recently reported that BMP-3B inhibits tumor formation of mammary tumor cells by promoting phosphorylation of Smad3[40].
An important finding of our study is the identification of mechanism where Runx2 protein downregulates BMP-3B levels by interacting and recruitment of Suv39h1 methyltransferase at the proximal regulatory sequence. Similar to our findings, a direct interaction of Suv39h1with the C-terminal domain of other Runx family members (Runx1 and Runx3) results in silencing of CD4 gene by promoter methylation during T-cell development [41,42]. Runx2 is well known to regulate chromatin structure and modulate target gene expression [43]. For example, Runx2 interaction with p300 alters chromatin structure (acetylation of histones H3 and H4) during activation of MMP-13 gene in bone cell lineage in response to PTH [44] and enhances histone acetylation resulting in increased Snail expression and decreased E-cadherin in lung cancer cells [33]. Recent reports indicate that Runx2 forms complexes containing the RNA Pol I transcription factors UBF1 and SL1, co-occupies the rRNA gene promoter with these factors in vivo, and affects local chromatin histone modifications at rDNA regulatory regions during rDNA suppression [45,46]. Consistent with these studies, our results revealed that Runx2 regulates histone H3K9 methylation status of BMP-3B promoter in lung cancer cells. There is a possibility that Runx2 repressor complex on BMP-3B promoter includes members of HDAC family as previously shown for repressing bone sialoprotein gene expression in osteoblastic lineage cells [47-49].
In summary, our study demonstrates BMP-3B as a novel target gene for Runx2 in bone lineage and lung cancer cells and provides insight into mechanisms that regulate epigenetic silencing of tumor growth inhibitors in lung cancer cells (Figure 5). Further studies are required to definitely establish the contribution of Runx2 in lung cancer progression.
Figure 5. Illustration of Runx2-mediated epigenetic silencing of BMP-3B in lung cancer cells. The recruitment of Runx2 and Suv39h1 on the BMP-3B locus increases histone H3K9 methylation status resulting in downregulation of BMP-3B expression in lung cancer cells.
Taken together, our results identified BMP-3B as a new Runx2 target gene and revealed a novel function of Runx2 in epigenetic silencing of BMP-3B in lung cancer cells. Our studies with modulation of Runx2 levels in lung cancer cells indicate that Runx2-mediated downregulation of BMP-3B levels is via interacting with methyltransrefase Suv39h1 and increasing histone H3K9 methylation status of the proximal promoter. These results suggest that Runx2 is a potential therapeutic target to block tumor suppressors gene silencing in lung cancer cells.
Materials and methods
Cell Culture and treatments
Normal bronchial and lung fibroblast (NL-20; WI-38, and IMR-90) and lung cancer cells (small cell lung cancer and non-small cell lung cancer cells: H720, and H1299) were cultured in growth medium as specified by American Type Culture Collection. The construction and procedure for wild type Runx2 or DNA binding mutant expressing adenovirus and lentivral transduction in normal and cancer cells are reported previously [24,45].
Animal procedures
Animals were maintained at the University of Massachusetts Medical School following procedures approved by the Institutional Animal Care and Use Committee (IACUC). Primary calvarial cells from Runx2 -/- mice were isolated as previously described [24].
shRNA treatment
Normal bronchial NL-20 or lung cancer H-1299 cells were transduced with lentivirus expressing shRNA-Runx2 target sequence 5'-AAGGTTCAACGATCTGAGATTTG-3' sequence in pLVTHM vector under H1 promoter [34]. Runx2 knockdown efficiency was confirmed by western blot and real time RT-PCR analysis.
Western blot analysis
Runx2 protein levels in normal bronchial, fibroblast and lung cancer whole cell lysates or nuclear lysates were detected by western blot analysis as described previously [24]. Runx2 antibody (MBL Inc., Woburn, MA) or Suv39h1 (C-14, Santa Cruz Biotechnology Inc. Santa Cruz, CA) and HRP-conjugated secondary antibodies (Santa Cruz) were used to detect immunoreactive proteins.
Chromatin immunoprecipitation
Chromatin immunoprecipitation (ChIP) was performed as previously described [34]. Protein-DNA complexes were immunoprecipitated using Runx2 antibody (M-70, Santa Cruz Biotechnology Inc.), Suv39h1 (Santa Cruz Biotechnology Inc.) and histone H3K9 (Abcam, Cambridge, MA) or IgG as a control. Purified DNA was subjected to real time PCR amplification with SYBR Green chemistry on an ABI real time thermocycler. BMP-3B promoter fragment containing Runx elements were amplified using forward primer: 5' ACT TTG ATG AAT CCG CAA CC-3' and reverse primer: 5' TTG TCT TGC CTC TAGCAG GAT-3'.
Real time RT-PCR analysis
The mRNA levels of Runx2, BMP-3B, GAPDH and 28S in primary osteoblasts, normal lung fibroblast, bronchial and lung cancer cells were analyzed after adenovirus- or lentiviral-mediated Runx2 transduction. Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's specification. Purified RNA was oligo dT primed and cDNA synthesized at 42°C with SuperScript II RNA polymerase (Invitrogen). For PCR amplification, the following primers were used: Runx2, forward primer: 5'- CGG CCC TCC CTG AAC TCT -3', reverse primer: 5'- TGC CTG CCT GGG GTC TGT A -3', GAPDH, forward primer: 5'- ATG TTC GTC ATG GGT GTG AA -3', reverse primer: 5'- TGT GGT CAT GAG TCC TTC CA -3'. BMP-3B, forward primer: 5'-AGC TGC TGG ACT TTG ACG AG-3', reverse primer: 5'-TGA CAA TGC TCT GGA TGG TG-3'. 28S, forward primer: 5'- GAA CTT TGA AGG CCG AAG TG-3', reverse primer: 5'-ATC TGA ACC CGA CTC CCT TT-3'. The gene expression levels were quantified by ΔΔCt method of relative quantification by normalizing the data with internal control and expressed relative to appropriate control cell line as indicated in the figure legends.
Wound healing assay
H1299 cells stably expressing Runx2 or empty vector treated control cells were cultured in triplicates in a 6 well dish with reduced serum conditions (0.2% serum) for overnight. The next day, a scratch was made approximately in the center of the monolayer by a sterile 200μl pipette tip. The detached cells and debris were washed with serum-free RPMI medium. The cells were then supplemented with or without TGF-β (5 ng/ml) containing RPMI medium. Five random images per well were photographed at 0h, 6h, 24h and 48h. The distance of the scratch was measured in ImageJ software at every time point. The wound distance at 0h was assigned as 100% and used to calculate percent wound closure at other time points. The P-value for statistical significance was calculated by unpaired T-test.
Cell proliferation assay
H1299 cells stably expressing Runx2 or empty vector treated control were counted in a hemacytometer and 1000 cells per well were seeded in a 96-well plate. To determine the changes in proliferation, the cells were indirectly assayed for cell number via a tetrazolium compound-based colorimetric assay (CellTiter 96 kit from Promega Inc. Madison, WI) according to manufacturer's instructions. At indicated time points over a period of four days, the cell titer reagent (20μl/well) was added to the plate and incubated at 37°C for 1 hour. The quantity of color developed (formazan product from the tetrazolium compound) was measured by reading absorbance at 490 nm in a spectrophotometer (Fluostar Optima BMG Labtech Inc. Cary, NC).
Lung cancer H1299-WT-Runx2 or -shRunx2 cells were washed with ice-cold PBS and harvested in lysis buffer [50 mM NaCl, 50 mM Tris (pH 8.0), 1% NP- 40, 25 mM MG132, and 1× protease inhibitor mixture (Roche, Indianapolis, IN)]. Lysates were incubated overnight at 4°C with 3 μg of rabbit antibodies against Runx2 antibody (M-70, Santa Cruz Biotechnology Inc.), and Suv39h1 (Santa Cruz Biotechnologies). Lysates were then incubated with protein A/G beads for 2 h, followed by four washes with wash buffer [50 mM NaCl, 20 mM Tris (pH 8.3), 0.5% Na-deoxycholate, 0.5% Nonidet P-40, 2 mM EDTA, 25 mM MG132, and 1× protease inhibitor mixture]. The total cell lysates and immunoprecipitated protein complexes were resolved by 8% SDS/PAGE and transferred to polyvinylidene difluoride membranes (Immobilon-P, Millipore, Billerica, MA). Blots were incubated with Runx2 (M-70) or Suv39h1 (C-14) antibodies. Membranes were then incubated with HRP-conjugated secondary antibodies against rabbit or mouse (1:2,000). Proteins bands were visualized with a chemiluminescence detection kit (Perkin–Elmer Life Sciences,Waltham, MA).
WI-38 and H1299 cells grown on gelatin coated cover slips were processed for immunofluorescence microscopy as previously described [31] using rabbit polyclonal Runx2 antibody (Santa Cruz Biotechnology, Inc.), followed by incubation with Alexa 488 conjugated secondary antibody (Molecular Probes, Eugene, OR). All images were taken using a Zeiss Axioplan digital microscope and analyzed using Metamorph software (Universal Imaging, Downingtown, PA).
BMP-3B, Bone morphogenetic protein-3B; DBD, DNA binding domain mutant; shRNA, Small hairpin RNA.
Manish Tandon, Karthiga Gokul and Syed A Ali contributed equally to this work.
Competing interest
The authors declare that they have no competing interests.
Author's contributions
MT and KG generated stable cell lines and analyzed cellular and molecular effects of Runx2 overexpression or knockdown (proliferation assay, wound healing assay, real time RT-PCR analysis, Western blots); SA performed ChIP and co-immunoprecipitation assays; ZC performed real time RT-PCR analysis and Western blots of samples with Runx2 overexpression or knockdown; JL and GS participated in the data analysis; JP conceived and coordinated the study and wrote the major part of the paper. All authors read and approved the final manuscript.
The authors would like to thank Dr. Jeffrey Borgia, Rush University, for providing bronchial epithelial NL-20 cell line, and Drs. Janet Stein and Andre vanWijnen, University of Massachusetts Medical School, and Dr. Carl Maki, Rush University Medical Center for stimulating discussions throughout the study. Studies reported were supported by National Institutes of Health grant 5 R03 CA123599-02 (JP). The contents of this manuscript are solely the responsibility of the authors and do not necessarily represent the official views of the National Institutes of Health.
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Oncogene 2004, 23:3521-3529. PubMed Abstract | Publisher Full Text
Kraunz KS, Nelson HH, Liu M, Wiencke JK, Kelsey KT: Interaction between the bone morphogenetic proteins and Ras/ MAP-kinase signalling pathways in lung cancer.
Br J Cancer 2005, 93:949-952. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
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Bannister AJ, Zegerman P, Partridge JF, Miska EA, Thomas JO, Allshire RC, Kouzarides T: Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain.
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Science 2001, 292:110-113. PubMed Abstract | Publisher Full Text
Choi KY, Kim HJ, Lee MH, Kwon TG, Nah HD, Furuichi T, Komori T, Nam SH, Kim YJ, Kim HJ, Ryoo HM: Runx2 regulates FGF2-induced Bmp2 expression during cranial bone development.
Dev Dyn 2005, 233:115-121. PubMed Abstract | Publisher Full Text
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J Cell Biochem 2003, 88:446-454. PubMed Abstract | Publisher Full Text
Javed A, Barnes GL, Pratap J, Antkowiak T, Gerstenfeld LC, van Wijnen AJ, Stein JL, Lian JB, Stein GS: Impaired intranuclear trafficking of Runx2 (AML3/CBFA1) transcription factors in breast cancer cells inhibits osteolysis in vivo.
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Cancer Metastasis Rev 2006, 25:589-600. PubMed Abstract | Publisher Full Text
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J Biol Chem 2011, 286:37335-37346. PubMed Abstract | Publisher Full Text
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Breast Cancer Res 2010, 12:R89. PubMed Abstract | BioMed Central Full Text | PubMed Central Full Text
Baniwal SK, Khalid O, Gabet Y, Shah RR, Purcell DJ, Mav D, Kohn-Gabet AE, Shi Y, Coetzee GA, Frenkel B: Runx2 transcriptome of prostate cancer cells: insights into invasiveness and bone metastasis.
Mol Cancer 2010, 9:258-275. PubMed Abstract | BioMed Central Full Text | PubMed Central Full Text
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Taniuchi I, Osato M, Egawa T, Sunshine MJ, Bae SC, Komori T, Ito Y, Littman DR: Differential requirements for Runx proteins in CD4 repression and epigenetic silencing during T lymphocyte development.
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Chakraborty S, Sinha KK, Senyuk V, Nucifora G: SUV39H1 interacts with AML1 and abrogates AML1 transactivity. AML1 is methylated in vivo.
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Lian JB, Stein JL, Stein GS, van Wijnen AJ, Montecino M, Javed A, Gutierrez S, Shen J, Zaidi SK, Drissi H: Runx2/Cbfa1 functions: diverse regulation of gene transcription by chromatin remodeling and co-regulatory protein interactions.
Connect Tissue Res 2003, 44(Suppl 1):141-148. PubMed Abstract
Boumah CE, Lee M, Selvamurugan N, Shimizu E, Partridge NC: Runx2 recruits p300 to mediate parathyroid hormone's effects on histone acetylation and transcriptional activation of the matrix metalloproteinase-13 gene.
Young DW, Hassan MQ, Pratap J, Galindo M, Zaidi SK, Lee SH, Yang X, Xie R, Javed A, Underwood JM, Furcinitti P, Imbalzano AN, Penman S, Nickerson JA, Montecino MA, Lian JB, Stein JL, van Wijnen AJ, Stein GS: Mitotic occupancy and lineage-specific transcriptional control of rRNA genes by Runx2.
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Ali SA, Zaidi SK, Dacwag CS, Salma N, Young DW, Shakoori AR, et al.: Phenotypic transcription factors epigenetically mediate cell growth control.
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Westendorf JJ: Transcriptional co-repressors of Runx2.
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Lamour V, Detry C, Sanchez C, Henrotin Y, Castronovo V, Bellahcène A: Runx2-and histone deacetylase 3-mediated repression is relieved in differentiating human osteoblast cells to allow high bone sialoprotein expression.
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Crit Rev Eukaryot Gene Expr 2011, 21:101-113. PubMed Abstract | PubMed Central Full Text


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We take for granted today's speed of banking. It is literally the speed of electrons—funds are wired across continents in moments, communication and transactions over distances are approved the same day, and complex documents are quickly certified with electronic signatures. Not long ago, however, international business moved at a much slower pace.
Adolph Sutro was a twenty-year old emigrant when he arrived in the United States in 1850. A decade later he followed the "call to Washoe" and ventured over the Sierra to the new mining areas in Nevada. He researched the Comstock and noticed that the primary mines started high up on the face of Mt. Davidson and their shafts quickly dropped into the mountain. Traditional mining relied on pumps to dewater the mines, but Sutro preferred to burrow a tunnel from a canyon below that would let gravity drain the shafts. To cover the costs of the tunnel, Sutro proposed that the mines pay the tunnel a royalty for drainage. The idea was met with skepticism.
The tunnel was eventually built. But it took five years to form the Sutro Tunnel Company and another 13 years of construction until it finally intersected with mine shafts on the Comstock. The tunnel consumed large amounts of capital, so much so that Sutro needed overseas financing. French capital was blocked by the Franco-Prussian War, so Sutro raised money through McCalmont Brothers, in the heart of the London Financial District. McCalmont raised both equity and debt funding for the tunnel.
After Sutro's tunnel linked with Comstock mines, Sutro wanted to re-aquire shares from McCalmont, and sell them in the US. He sent a letter in September, 1879, requesting the transfer; I recently came across the response from the London bankers. Note the drawn out timeline of the transaction: Sutro sent his request in September and the reply was sent in late October, a reply stating that the transfer would take more time since it required the physical partner signatures on the certificate of shares. The letter was sent Royal Mail to Sutro, across the Atlantic via steamship. Sutro received the letter in November. This simple transaction took more than three months to complete.
The letter that documents this tedious transaction was sent from McCalmont's headquarters at 15 Philpot Lane, London. Today this location is mere blocks from Wells Fargo Bank's currentLondon banking offices at Plantation Place, 30 Fenchruch Street. A similar transaction of share transfer would take so much less time today, depending on where shares are registered, Compared to the months it took for Sutro's request, imagine what our customers can do today with current technology!

subset: commoncrawl


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Viewing convention Program information from 2008
Session Details
Monday, 29 December
613. Core Issues in Language Theory
3:30–4:45 p.m., Nob Hill D, Marriott
Program arranged by the Division on Language Theory
Presiding: José Alberto Elías-Ulloa, Stony Brook Univ., State Univ. of New York
1. "Word-Internal Voicing Neutralization in Duan Zhuang," Seunghun J. Lee, Rutgers Univ., New Brunswick
2. "Cliticization and Its Discontents," Eric J. Hyman, Fayetteville State Univ.
3. "Creative Verb-Noun Compounds in Spanish: A Cognitive Perspective," Jiyoung Yoon, Univ. of North Texas
No comments have been posted for this session.
|© 2015 Modern Language Association.


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on 02-02-2010 08:17
hi all , wonder if anyone can help ? my tv picture keeps flashing green it started about 2 weeks ago went away and now its back lol im using a standard virgin box connected to tv with scart , ive done all the norm stuff ie.. turned box off left it 20 secs and turned back on , checked scart lead checked smart card , checked virgin cable wire etc etc , checked tv with norm analogue ariel and picture is fine , anyone else having these problems ? plz help plz help lol cant bare another phone call to vm after still no internet since begining of dec09 and also there just gonna talk to me like im 9 years old do this do that when ive already done it all oh boy...
anyway thxs in advance
on 02-02-2010 08:22
I don't want to sound pedantic, but have you checked your v+ box with another Scart-enabled TV?
on 02-02-2010 09:25
hi and thxs for the reply ! yes ive tried everything i guess im just tryin my luck here before ringin vm up im clutchin at straws because i really cant stand ringin them anymore ! thxs again
on 17-03-2010 13:11
This is 100% a SCART issue, VM will be unlikely to help with this, you need a new scart cable, otherwise it is the scart socket on the back of the tv, it could also be the scart socket on the virgin box, but that is very unlikely. You can fix this temporarily buy going into the virgin box setttings, and changing the scart output to composite, (in the vbox this is settings, 4, not sure about v+ but will be similar) in this mode you picture will be in full colour, but not as good quality and much less sharp.
I recommend getting a new scart cable (an RGB compatable one), but is it does turns out to be the socket on the back of the tv then there is not much you can do about it, even if you have 2 scart sockets on your tv, the spare scart socket will not support RGB anyway, so it will be no better that the temporary solution i gave you above.
on 14-05-2010 20:49
My TV sometimes flashes between red and blue. Tried asking 'Jenny' but no use at all. I eventually solved this myself by pressing the reset button on the V-box.


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Don't get too excited. The new trailer for Showgirls Exposed, the sequel to Showgirls, is awful. Sure, there's neon lights and ass, but I can give you a list of better ways to see such a combination. Well no, not right now. Ask me later, when I'm NOT writing a post.
It appears as if director Marc Vorlander grabbed a Handycam, a bottle of peach schnapps, and stumbled drunkenly through a nondescript city filming debauchery. If you stumble around long enough, chances are you're going to capture girls having a three-way with a dog, a stripper getting bludgeoned, and a mob boss executing one of his minions. In fact, I saw all three such things the other night whilst walking through a parking lot in Hollywood. I was British that evening.
Check out the NSFW trailer for Showgirls Exposed after the jump…IF YOU DARE.
Consider posting a comment.


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Could you imagine if every ABA graduate were assessed on the same level as Mitchell was in Mass? How many would not be allowed to take the bar in their state? Either say the education is good or not, but to say it really depends on your grades, your participation in extracurricular, how quickly passed the bar, shows a complete discord.
Screw it, go to SCUPS. Plus, yeah dude we know you're talking about yourself in the third person. It is so tiresome to have school recruiters pretend to be students on this board. Yeah, I see you.....
Good. Online grads shouldn't be admitted anywhere.
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Tuesday, September 21, 2010
© Copyright 2015 Clayton News Daily
By Joel Hall
After six years of planning, the City of Lake City soon will have its own 29-acre nature preserve. With the help of the Clayton County Water Authority (CCWA), Jester's Creek will run through the area, as it once did several decades ago.
On Tuesday, Lake City and water authority officials broke ground on the Lake City Nature Preserve and East Jester's Creek Stream Restoration Phase 4 project.
The Lake City Nature Preserve, scheduled to be completed within one year, will sit on 29 acres of land behind the Lake City Municipal Center. According to Ron Gossett, project manager for the preserve, the large-scale nature park will include 1.7 miles of walking trails, a man-made lake with a waterfall feature, pavilions, and observation areas.
"There's been so much construction over the last 15-to-20 years that they've [city officials have] seen all of the city's wooded areas bulldozed," Gossett said. "This place grew up in the 1960s, and 1970s, and we didn't conserve much of the land. This was their last chance to reclaim some of that. It adds to the quality of life to be able to get out into the country, but still be in town," he said. "We hope to provide them with a quiet place to get out and enjoy nature."
Included in the project is the restoration of 1,650 linear feet of East Jester's Creek. Since 2001, the water authority has restored natural water flow to three other sections of Jester's Creek, one located outside of Riverdale and two located within the city limits of Morrow, according to CCWA Stormwater Manager Kevin Osbey.
"This started with our watershed improvement program that started in 2001," Osbey said. "In that study, we found different stream sections that were impacted. We saw more construction, and with more construction, more water was introduced into the stream. As more water began hitting the stream, you saw more erosion.
"On the stream banks you saw a lot of silt deposits," Osbey continued. "What we want to do is put the meanders back in, and slow down some of the velocity. We want to restore the banks again by planting trees.
"You don't want your city, or county, to turn into New York City, where there is a building on every space in the county," Osbey added. "Knowing that you can come into a 27-acre greenspace, where no one can develop, it can only be a good thing."
According to Gossett, the total cost of the nature preserve and creek restoration project is approximately $2.4 million, which is being funded through Special Local Option Sales Tax (SPLOST). He said Lake City will partner with the Reynolds Nature Preserve in Morrow, to eventually designate the Lake City Nature Preserve as an official bird sanctuary.
Lake City Mayor Willie Oswalt said that within the next three years, the city also plans to build a new community center and a new public works building using SPLOST dollars. He said the nature preserve will serve as "another star" for Lake City.
"This is not only good for Lake City, it's good for the county," said Oswalt. "You will see people come here just to eat lunch. It'll be an upgrade for a lot of people."
More like this story
- Morrow passes stream restoration funds ( June 26, 2003 )
- Lake City to get new public works building ( October 13, 2010 )
- Morrow begins Jester's Creek trail's second phase ( September 4, 2009 )
- City officials share economic development plans ( January 27, 2011 )
- Volunteers sought for clean-up effort ( October 6, 2010 )


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Vance & Hines Shortshots Original Exhaust For Harley FXR
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Get up to speed with the latest from LVGP!
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2/24 Team 333 & HGR take Top Honors at LVGP500
The Daytona 500 signifies the start of the racing season. It is always an exciting time of year for the racing community. To celebrate, Lehigh Valley Grand Prix started the LVGP500 4 years ago. This exciting race is a 500 […]Read More
11/18 Congrats to ALL the Participants at LVGP's 1st Turkey Bowl!
The Turkey Bowl was a combination of high speed, wheel to wheel karting action and an opportunity for the racing community to give back to those who are less fortunate. Racer's donated canned goods and non-perishable food items for the […]Read More
2/7 Dramatic Finish to Adult Leagues
Group B Provides Closest Points Race in LVGP League History! It was another exciting round of leagues at LVGP. The final week the field had the opportunity to run the league races on the new Super Karts which add a […]Read More


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AURORA, ONTARIO, JULY 9, 2007 - For the opportunity to live in an outstanding location, in a spectacularly designed home packed with luxurious standards and outstanding value – hurry to Great Gulf Homes' neighbourhood at River Ridge in Aurora.
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Psalm 37: 5-6 "Commit your way to the LORD, Trust also in Him, And He shall bring it to pass. He shall bring forth your righteousness as the light, And your justice as the noonday."
Sometimes promises take longer to eventuate than we would like, but for good reason - and only the Lord knows those reasons. He knows what is best. As a reward for our patience, trust and faith, our righteous reward is coming. And as a result of us waiting on Him and holding firm and steadfast and not letting go of His hand, through our suffering God is getting the glory and we are being led to certain victory. So, do not give up - ever!


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Store makeovers and hidden cameras. Just a couple of ways food coops are trying to keep big organic grocers from muscling in.
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eMoney News
Vending machines to change in Grand Canyon National Park
29 Dec 2011
The vending machines situated within the Grand Canyon National Park are to undergo some changes in order to make them more environmentally-friendly.
As of early 2012, users will no longer be able to purchase disposable plastic bottles of beverages as part of a plan by park officials to keep the ecosystem sustainable.
There is also to be a review of water availability, health and safety by the National Park Service (NPS).
Officials have already been trying to get visitors to fill their own reusable canisters with water to save on plastic, but it is hoped this new move will cut down litter and the use of natural resources even more.
Currently, 30 per cent of park waste is made up of plastic water bottles.
According to the NPS, almost five million people see the mile-deep Grand Canyon every year, with the majority overlooking it from the south rim.
Park authorities could consider bringing in vending machines that use contactless smart cards for payment, as it may cut down on maintenance and reduce the likelihood of vandalism occurring.


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Last Updated: February 7, 2013 09:54am ET
Logistics Center "Ready for Build-to-Suit"
STOCKTON, CA-Subdivision maps are being processed to accommodate greater lot-line flexibility by mid-2013 for the newly rebranded industrial park, NorCal Logistics Center.
You can now be notified via email if this story is updated by clicking on the "Follow this Story" link. You must be a registered member to take advantage of this "members only" benefit.


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The Blog
Arianna in Israel
3:01 PM, Oct 1, 2009 • By RACHEL ABRAMS
Mrs. Huffington visits Israel -- a lifelong dream -- and a disappointingly boring series of sketches is all she manages to wring out of her visit. The "progressive populist," who laughably tells the Jerusalem Post that "At The Huffington Post we really avoid looking at American politics with the prism of Right and Left because we believe it is very obsolete and it obscures more than it reveals," spends most of her time in the Holy Land talking to like-minded people -- that is, people who, with her, find Israel as culpable in its ongoing struggle against anti-Semitic Islamofascism as its mortar-shooting bus-exploding cafe-bombing terrorizing neighbors. When her interlocutors deviate from Israel-is-to-blame orthodoxy, she makes sure to let them know -- more perhaps in sorrow than in anger, but still.
A few highlights from an otherwise pedestrian journal:
Israeli President Shimon Peres, extolling the intellectual resources that have made possible his country's astonishing scientific and technological achievements, gets this amazing rejoinder: "But, I countered, brains without heart and empathy are never enough. After all, Germany in the 30s was a highly educated country."
To which the neither-Right-nor-Left Mrs. H. responds: "There is an enormous amount of philanthropy in Israel but the horror of what happened to civilians, especially children, in Gaza continues to overwhelm the good that's been done."
She visits the Wailing Wall, her shoulders covered "with a hastily borrowed shawl," places her own "folded up prayer" in a crevice and wonders, "What is it about shoulders, in particular, that God would find so disquieting?"
She dines with Ehud Barak, asks him "to compare George W. Bush's leadership to Obama's when it comes to Israel," elicits the really very indiscreet: "'I'm an ABB,' he said. 'Anyone But Bush.'"
She enjoys the hospitality of Rabbi Daniel Gordis at lunch, and listens, apparently without remark (she doesn't make one here, at any rate) to his impassioned arguments against the Goldstone Report -- which is as far as she ever ventures from her blame-Israel comfort zone; if she visits any settlers, for example, or speaks to a single representative of the Israeli Right, she doesn't say -- then trots off back to more familiar territory for a succession of meetings with her favorite roster of Palestinian victimologists.
And oh, the sorrow and the pity: "The security wall. The roadblocks and barbed wire. The separate roads that the Palestinians have to use. The checkpoints and 'buffer zones.' The very large, sprawling, and very permanent-looking Israeli settlements carved out on Palestinian land. No wonder Palestinians feel like strangers in their own land."
Yes, maybe that's why, Arianna, or maybe it's because by their murderous refusal to accept the existence of the Jewish Homeland they have relinquished any claim to sharing any part of it.


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How to Get Dried Masking Tape off a Painted Surface
You can remove any adhesive with a solvent. The real question is, which solvent is best to use?
1Determine what kind of painted surface you have. If it's the interior of a house, then you can be pretty sure that it's latex paint. A product called "Goof Off" is available in home stores (paint department). There are some citrus-based solvents available too, generally available in the same area.Ad
2Scrape off what you can of the tape using a fingernail or a putty knife. Be careful not to scratch the painted surface. A nylon pancake spatula might work too. You have to try to get the paper part off first so that the solvent can work on the glue.
3Prepare for adding the solvent. Goof-Off is going to dissolve both the adhesive and the paint, so work carefully. Put a little on a rag, rub it on the adhesive just until it's removed. Then wipe off the remaining solvent with another clean rag or paper towel.Ad
Give us 3 minutes of knowledge!
- Always read the manufacturer's instructions on the solvent product.
- You can ruin a lacquer finish (furniture, automobiles) with solvents. Test first!
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The Bonus – Should I Stay or Should I Go? View Count: 307
When evaluating when the right time is to make a move, there are many factors that come into play, one of them being your year-end bonus. This article is going to break up the year into quarters and guide you through your options at different parts of the year in relation to your year-end bonus. Read More at BCG Attorney Search
The Bonus - Should I Stay or Should I Go? by Scott


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Sometimes I do, sometimes I don't.
If you live in a suburb with ridiculously low crime-rates, it's day-time, and you're at home with several people, and still feel the need to have your doors locked for safety, I'd probably call you paranoid.
Reversely, if you live in the inner-city, in an apartment, and don't lock your door, regardless of time and who's with you, I'd call you a risk-taker.
There's a time and place for everything.


text:
The Victorinox 40085 Fine Cut Round Steel is a complementary companion to any kitchen cutlery set. In order to get the best performance from your blades it's essential to keep them sharp, as a dull blade leads to an inferior end product and unwanted slips. The best way to avoid these mishaps is to have a good sharpening steel handy for honing, straightening, and re-sharpening the blades you use most to ensure they're working properly. And what better tool to keep your blades at their highest quality than the Victorinox 40085 Fine Cut Round Steel! Measuring 12 inches from tip to handle, this sharpening steel is large enough to accommodate any sized blade. The fine cut means this steel has a moderate bite and is great for smoothing and straightening the edge of your blade. Comes equipped with a classy dark brown wooden handle and a loose ring on the end for easy handling and hanging!
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Our sister site Collectible Cars of Arizona sells restored and original Vintage & Classic American Automobiles, from clean drivers to museum-quality restorations (and we can ship worldwide). Click on the link or screenshot image below to view our current classic car inventory, or call Jerry at 602-253-8081 for more information.
Design: the Way Home media


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RUSSELL, TEXAS (Van Zandt County). Russell, twelve miles northwest of Canton in northwest Van Zandt County, was a switch on the Texas and Pacific Railway and later the site of an abandoned railroad station. The town was named for Eric Russell, who owned the site. The Memorial Methodist Church, founded in 1875, was destroyed by a tornado and rebuilt in 1890. A population of ten recorded in 1930 had postal service from Wills Point, but by 1936 only scattered dwellings remained. Subsequent state highway maps no longer listed the community.
The following, adapted from the Chicago Manual of Style, 15th edition, is the preferred citation for this article.Diana J. Kleiner, "RUSSELL, TX (VAN ZANDT COUNTY)," Handbook of Texas Online (http://www.tshaonline.org/handbook/online/articles/hvr77), accessed October 09, 2015. Uploaded on June 15, 2010. Published by the Texas State Historical Association.


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Details about Gold Diamond gemstones Suction Dredge 8-inch AP 8226See original listing
Item condition:
New: Never Used
20 Jul, 2014 17:28:31 AEST
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New: Never Used: A new, unused item with absolutely no signs of wear. The item may be missing the original packaging, ... Read moreabout the condition
for sale : Suction Dredge 8-inch AP8226 with magnetic recovery system for Gold, Diamonds and Gemstones mining.
Suction Dredge with MAGNETIC recovery system for gold, diamonds and gemstones mining. Ordinary riffles accumulate heavy black sand, most of content are magnetic minerals. Magnetic minerals have specific gravity about 4.9-5.2g/cm3, and push out valuable gemstones( SG of ruby and saphires -4.0g/cm3, SG of diamonds- 3.5g/cm3). In our dredges and sluices we use NbFeB Permannent Strong magnetic field ( B=1.5 Tesla), which removes all stong and week magnetic minerals. This also improves finest gold recovery due to fact 80% of concentrate is magnetic and pre-removed from riffles.
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In this model of dredge used only aluminum and stainless steel parts are designed for ease of assembly long last and with total weight 230 kg only.
Model AP8226
Engines 188F Electric start 2x13 hp
Pumps: Pressure 75m
Capacity 75m3/hour
Power Jet: 8" S/steel 1.2m with 2 logs 3"
Suction tip : 8" S/steel
Air Compressor : 12v electric
volume -150L/min
Float Dimension 1.7x3m
Pontoons, each:
weight 6kg
size D0.45mx3m
Buoyancy maximum each pontoon 400kg
Suction Depth 7m
Fuel Consumption 8l/h
Capacity 25 m3 per hour
Sluice Box 0.2x0.71x2.3m
Hose Length 5m
Net Weight 210kg
Shipping Weight 220kg
1. Personal & Business Checks (Subject to 10 Banking Day hold)
1. Personal & Business Checks (Subject to 10 Banking Day hold)
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2. Fuel supply-0.00
3. Life time of brushless motors many times higher compared to fuel engines.
4. Service and spare parts-0.00


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Helicopter Parents Heading for a Crash
Hovering over kids' lives isn't good for either child or parent.
November 2, 2008 - 12:30 am
With the school year well under way, helicopter parents are spinning into overdrive, hovering over their children and micro-managing their lives. But where did it come from, this belief that good parenting is synonymous with doing everything for one's child? What makes so many otherwise sane and rational parents believe they need to confront a "mean" teacher to demand a better grade for their child? To rally against school bans on the cell phone they gave their kid so they could keep in touch throughout the day? To fill up a child's schedule with piano lessons, soccer, football, Scouts, and additional tutoring, then spend their evenings chauffeuring them to each?
Part of it no doubt stems from the constant awareness that other parents are watching — and closely. Ever since Hillary Clinton bellowed "it takes a village to raise a child," well-meaning parents have bought into the notion that they aren't simply responsible for looking out for their own children's well-being, but for that of others' kids, too.
Did Johnny's friend show up for a sleepover hungry and wearing dirty clothes? Perhaps it would be best to grill him ask if his parents are having marital or substance abuse problems that stand in their way of providing for their child. (Because surely it couldn't be that the boy's mother served salad for dinner, which he refused to eat, and objected to washing a special load of clothes after her son forgot to bring his hamper to the laundry room.) Why does Lakeisha have a cast on her arm so soon after getting off crutches from her sprained ankle? Sure, Lakeisha swears both injuries are from her Saturday soccer games, but perhaps someone ought to ask Child Services to look in on the family "just to be safe."
Ever aware that someone else might be looking at one's own kids and jumping to the worst possible conclusion, many parents have grown accustomed to thinking of child-rearing as a public performance. A parent who shrugs her shoulders when her child comes home with an F on a math exam — when he's clearly capable of doing so much better — must not care , or so an outside observer might conclude. Even if Mom knows Joey was playing video games in his room when he should have been studying, and that living with an F is a good way for him to learn about the consequences of bad choices, she still worries what other people will think . Why deal with that kind of condemnation, or expose Joey to it, when it's so easy to demand a retest and assume Joey learned his lesson already.
Part of the blame also lies with the school system. For years we've been hearing about the importance of parental involvement in a child's education. Even the National Education Association urges parents to be involved not only in homework but within the classroom. After a decade of bombardment with statistics about kids falling through the cracks due to teacher shortages and crowded classrooms, good parenting has come to require ensuring one's own child doesn't get left behind. So is it truly that surprising when a parent, hearing that some involvement is good, assumes more involvement must be better?


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Call: 013397 55558
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You can now check availability and make a reservation online by clicking here, or by clicking the link on the left hand side of this page.


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shannen doherty
She's Back: Shannen Doherty Returns to TV with Wedding Reality Show
According to Entertainment Weekly, the show will follow Doherty and fiance Kurt Iswarienko as they plan their wedding. As if that wedding planning wasn't stressful enough, the show will follow Doherty as she renovates her home. Expect some mayhem.
"People will see a side of Shannen they never expected to see," John Miller, senior vice present of production and development at WEtv, said. Miller said Doherty was "unfiltered, honest and vulnerable" in a pitch tape.
Tiffani Thiessen and Matt Bomer Play 'Shaved By the Bell' on 'Watch What Happens' (VIDEO)
'White Collar' stars Tiffani Thiessen and Matt Bomer stopped by 'Watch What Happens Live' (weekdays, 11PM on Bravo) Thursday night and played a game of 'Shaved by the Bell' with host Andy Cohen. Pivoting off Thiessen's iconic '90s role as Kelly Kapowski on 'Saved by the Bell,' Cohen showed the pair a series of pictures of '90s stars with their heads shaved and Thiessen and Bomer had to guess who they were.
Despite the fact that everyone looked like Sinead O'Connor, Thiessen and Bomer fared pretty well, nailing Blossom, Screech, Shannon Doherty, Fabio, Alanis Morissette, Kato Kaelin, Vanilla Ice and Pauly Shore. They won a '90s mix CD and Bravo 'Mazel' shirts for playing. For some of the celebs featured in the montage, it was the most screentime they've seen in years.
Ricky Gervais, Will Arnett Book 'The Office' Season Finale and More Casting News
According to Entertainment Weekly, the episode is still being written so the roles are still taking shape.
Earlier this season Gervais reprised his role of David Brent from the UK version of 'The Office' and made a brief cameo opposite Steve Carell's Michael Scott. EW's source warned that viewers shouldn't assume Gervais or Arnett will play the new boss of Dunder Mifflin.
In other casting news ...
• Danny Bonaduce, Shannen Doherty and Barry Williams will star in new Syfy TV movies. Williams and Bonaduce will hunt Bigfoot and Doherty will star in 'Gretl' as the long lost sister of Hansel. Syfy is also prepping flicks about zombies and leprechauns. [TV Guide]
• Natasha Henstridge is heading to The CW's 'Secret Circle.' In the pilot, Henstridge will play the vice principal of New Salem High School, somebody who is familiar with the witch coven. [TVLine]
• Elizabeth Perkins will star opposite Michael Chiklis in CBS's 'Vince Uncensored.' The former 'Weeds' star will play Janet, a teacher and wife to Chiklis' Vince. The pilot follows Vince as he starts changing the way he lives his life after a medical problem. [Deadline]
Shannen Doherty Discusses Her 'Bad Girl' Past, the Miley Cyrus Bong Video and the Tabloid Media (VIDEO)
When asked why she thinks she was labeled as a bad girl, Doherty owned up to her behavior. "I think I made it a little easy for them," she admitted. "I think ... because ['Beverly Hills, 90210'] was such a sensation, that it was probably best to stay home, which I did not do."
She also indicated that she thinks the media in some ways dictated who they wanted to present as the bad girl, by looking at who was out and about the most of these super-successful young people. That theory might help explain, as the media and the Internet have made such videos and photos more prevalent, why there seem to be more celebrity "bad girls" in the past ten to fifteen years than we've ever seen before that.
Milo Ventimiglia, Shannen Doherty to Star in New Web Series About Sleep
The new series, 'Suite 7,' is sponsored by the Better Sleep Council, a division of the International Sleep Products Association. The goal of the seven-part series: To raise awareness of how important a proper night's sleep is for your health and well-being. Lifetime will air the series on MyLifetime.com, and it will also be available on Hulu.
Just who is going to teach you the importance of sleep? None other than 'Heroes' star Milo Ventimiglia and 'Beverly Hills 90210's' Shannen Doherty. But that's not all! Doherty's former costar Brian Austin Green will also appear, along with Craig Bierko, Illeana Douglas, Eddie McClintock and Jaime Murray.
Shannen Doherty Recalls Troubled Past, Weighs in on Lindsay Lohan (VIDEO)
Shannen Doherty on Being a Bad Girl and Her New Hallmark Channel Movie
But in her new movie, 'Growing the Big One' which premieres at 9PM ET, Oct. 23 on The Hallmark Channel, Doherty tackles a different kind of role, "the city girl who moves to the country."
She plays Emma, a young woman who inherits her deeply-in-debt family farm and decides to enter a pumpkin-growing contest in hopes of saving it with the winnings.
"I liked it because it's a lighter role for me," she told TV Squad. "I'm usually playing these darker roles, but I liked this one because it's lighter and fluffier. I'm an aunt and it's nice to finally do something that my brother's whole family can sit around and watch their aunt."
'Spin Crowd' PR Team Ruins Shannen Doherty's Store Opening (VIDEO)
Cheban says Doherty is "giving me the death glare of all death glares. She is scaring me." The visibly annoyed actress tells him, "You should've called me. I would've brought scissors from my house." Finally, the giant fake scissors arrive -- and "fake" is the appropriate word. They can't cut. Good job.
Shannen Doherty Talks 'Dancing' Elimination Conspiracy Theories (VIDEO)
"I have to tell you, after all the seasons we've done, you're elimination was the most surprising," Inaba told Doherty and her 'Dancing' partner Mark Ballas. Inaba was so shocked, she said she text messaged somebody asking, "How did that happen?" while she was on set. Regis Philbin, always quick with a comment, informed Inaba that it happened because "the judges didn't help them!"
Philbin tried to get the 'Beverly Hills, 90210' bad girl to gossip, but she wasn't falling for it. Bravo, Shannen.
Catch the video after the jump.
What You Missed Last Night: 'Lost' Secrets, Dunking Ninjas, Horse Singers & Starbucks
[Watch episode of 'Jimmy Kimmel Live' and other shows at SlashControl.]
Shannen Doherty Appeared on 'DWTS' for Stricken Dad (VIDEO)
Doherty's father, who suffered a near-fatal stroke on Christmas Day, congratulated his daughter for her short-lived run on the dance show. "My dad doesn't have his speech 100 percent back yet, but the first words he learned were, 'I love you.' And, so, he was crying and he said, 'I love you. I'm proud of you, baby,'" she said.
Watch the video after the jump.
'Dancing with the Stars' - 'Performance #1: Results' Recap
(S10E03) We start the first 'Dancing with the Stars' instantly by placing a couple in the bottom two. Pamela & Damien are in the bottom two, while Buzz & Ashly are safe. Instead of our traditional recap, Len critiques the dances in a recap fashion. Nothing out of the ordinary there.
Encore - Nicole & Derek's Jive - The second time around I think I absorbed the performance better. It's still too fast for my taste, and I wasn't fond of the slight Michael Jackson side-by-side choreography. I have the feeling that getting 10s early in the season will be her kiss of death.
While the show is now an hour earlier, the format of the results show didn't change. It basically is a clip, someone's safe, musical performance, someone's safe, repeat.
Shannen Doherty Sent Home on 'DWTS' (VIDEO)
"I think that dancing isn't really my thing," she said after the results were announced. "I'm an actor, and it's what I love to do. I did this for my dad, and I did exactly what I wanted." Plus, with her partner Mark Ballas injured, Doherty said she wouldn't have wanted to go on without him anyway.
We know she's had a "bad girl" image throughout her career, but 'DWTS' showed a sweeter side of Doherty that maybe we haven't seen before. That's always a good thing.
Watch the video after the jump.
'Dancing with the Stars' - 'Performance #1 Part 2' Recap
(S10E02) We continue 'Dancing with the Stars' with round two and the contestants performing the Foxtrot and the Jive. The order was shuffled this week with Kate Gosselin closing the show to an amazing train wreck ending. It was well worth having to sit through some painful performances.
It was interesting to see that the dancers who got the fast Cha-Cha last week got the slow Foxtrot, while the Viennese Waltzers got the Jive. It would have been really interesting to see how much bounce Pamela Anderson would have provided in the Jive, but I'm sure she'll survive until the second time Jive shows up again.
As the show started, I've gotta say how much I love that in the opening, Nicole Scherzinger's hair does the perfect whip. Years of Pussycat Dolls training did something.
Shannen Doherty on Why She's on 'Dancing With the Stars' (VIDEO)
Shannen Doherty appeared on 'George Lopez' (weeknights, 11PM ET on TBS) to talk about why she's on the show. It's a negotiation tactic. She made a deal with her dad that she'd do the show (which he loves) if he promised to work hard to come back from his stroke.
Sounds like a winner to us, even if Doherty gets booted off along the way.
Watch the video after the jump.
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Sensitive New Age Guys Never Ruled: Yoga, Existentialism & Staying Engaged in the World. ~ Eric Shaw
In my early 30s, I was doing something goofy in public that I thought was really cool, and someone called me a Sensitive New Age Guy.
The comment cut and caused soul-searching.
I manned-up more in the following years.
I haven't heard the term SNAG in awhile, but an allied concept, the "spiritual bypasser" now makes the rounds. It means you're avoiding life responsibilities of work and relationship by falsely "giving it all up to God."
Anyone—Sensitive or Insensitive, New Age, or Old Age—can avoid heroic engagement in the world.
That said, this morning—while immersed in the heroism of the long-avoided mopping of my apartment—all the while delaying my morning yoga practice—I felt I was doing the opposite. I felt I was "worldly bypassing."
The most heroic non-avoider of our new Yoga Nation is Gandhi. (I think we overlook America's Martin Luther King because he wasn't Hindu and he didn't do the whole gamut of yogi things that Gandhi did on his way to changing the world—avoid meat, live in an Ashram, consistently cite the Bhagavad Gita, etc.). Andrew Harvey's concept of "Sacred Activism"—mostly championed by women in the yoga universe—e.g. Seane Corn, Shiva Rea, Sharon Gannon—is an expression of this Gandhian ideal. Harvey says,
When the deepest and most grounded spiritual vision is married to a practical and pragmatic drive to transform all existing political, economic, and social institutions, a holy force—the power of wisdom, and love in action—is born.
This force I define as Sacred Activism.
Harvey's ideal wasn't yet articulated when I was a boy, but concepts like it colored my youth.
As I've said in other posts, my mother & father were both minsters, and the values of M. L. King (more than Gandhi) were very alive in my childhood home. I turned seven the year he was killed. King and other Christian Religious radicals were my boyhood heroes: The anti-war Catholic priests, Philip and Daniel Berrigan—who poured blood on nuclear warheads; the Yale Chaplain William Sloane Coffin who fought for gay rights; and the leftist theologians Richard and Rheinhold Niebuhr, who argued for a socially responsible nation.
Despite my family history, when I "got Buddhism" at the age of 25, meditation became more important than any worldly activity.
That year, I took up a long period of "spiritual bypassing."
Spiritual bypassing is a valid career option in India, so I had good company. The parivrajaka, the wandering yogi, has doubled down on the possibility that Realization results from dodging worldly action.
The philosophy of the Yoga Sutra can give us some guidance as to how to more intelligently orient our activity in the world. It divides Reality into spiritual and worldly realms—called Purusha and Prakriti. The realization of Purusha is the goal of the sramana, the yogi striver—a total spiritual bypasser—who seeks Enlightenment and has "tuned in, turned on and dropped out." On the other hand, the brahmana–the good everyday citizen who is still caught in the world's whirring wheels—keeps his floors mopped. If he's got a "bypass," it is of the worldly kind.
I was tempted to completely embrace a sramana path, but didn't do it. But the pull persists. For most of us, the pulls toward worldly action and spiritual practice stay in conflict.
In the Samkhya philosophy of the Yoga Sutras, Purusha, the realm of pure awareness, and Prakriti, the realm of action, remain eternally unresolved. The spiritual and worldly interact, but there is no holy One.
The Material and the Spiritual won't be reconciled by a collapsing universe or three Bethlehem wise men galloping to a cineplex to watch Apocalypse Now.
We will feel the pull toward life's contrary possibilities eternally.
The good news is this contrariness is dynamic.
It keeps us restless, provoked and alive.
I think I'm a queer duck in this equation, to use an old term. Even before my epiphany when I was 25, my boyhood saw me rolling my eyes up into my head for hours or resting at the bottom of swimming pools as long as I could, watching my mind get quiet while holding my breath.
I tend toward spiritual not worldly bypass, but I think most folks bypass the spiritual more than the worldly. America ain't no Nepal. Our ascetic ideal is expressed through the work ethic and—unlike India—a train ride to realization won't easily be bankrolled by street side begging. Society feels no obligation to the freelancer sramana.
In American Democracy, the obligation to be active in politics is more universal than any pull to meditate. But if you're in the yoga game, you are called to marry the aims of meditation and action. Activism is part of the work of self-transformation—even if you think meditation is enough.
It can be painful to intertwine action and self transformation, but—like childbirth or chaturangas, or getting out of bed in the morning—it's a pain we're made to endure, and it makes our inner and outer action potent. "Spiritual Bypassing" should be criticized because it is not whole.
Like Camus, we say the infinity of pushing our own personal rock uphill is a happy dilemma.
We feel pain on both sides of the equation—pushing into the world, or pushing toward the self. We are both angel and animal, and we ride the life's train in opposite directions.
"Spiritual Bypass" dodges transformation of the worldly "I."
"Worldly bypass" dodges realization of the Spiritual "I."
Sacred Activism tries to dodge these dodges.
About the time the Sutras were composed (200 CE),the Bhagavad Gita was, too. And here we get a nice formula—one championed by Gandhi (who read the Gita constantly). It allows us to be "worldly," but get spiritual goods, too.
The BG's description of Karma Yoga—acting in the world but giving the benefits to God—foreshadows "Sacred Activism." Karma Yoga accommodates our divided nature.
Meditation is usually called an otherworldly occupation, but when "the most grounded spiritual vision is married to a pragmatic drive"— to use Harvey's formulation—meditative goals come to worldly action.
Karma yoga and Sacred Activism are methods to change ourselves as we change the real world.
Kierkegaard said, "purity of heart is to will one thing." In Sacred Activism, even as we fight for variegated goals, we fight for the singular transformation of the spirit.
The Gita says, "Whoever works selflessly combines renunciation and activity. Right action is really renunciation." (6:1-2) .
This tack avoids either bypass. It swims consciously with Purusha and Prakriti.
Sacred Activism and Karma Yoga are methods for pushing the rock of the world forward and staying whole. Devoted to this game plan, life's pain is re-purposed and we, the human "animal-angel ," ride one bus toward divergent destinations.
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Editor: Kate Bartolotta
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7 Responses to "Sensitive New Age Guys Never Ruled: Yoga, Existentialism & Staying Engaged in the World. ~ Eric Shaw"
Really enjoyed this, Eric. Great reflections on an incredibly important topic, and beautifully written. Thanks!
This is a very well written and thoughtful piece. I appreciate its take on a very complex subject. I recently wrote about this dynamic myself in "So You Say You Want a Revolution? Well…".
For me though, your analysis perhaps misses the most subtle and important facet of this dilemma. And that is to point out that dedicating one's life wholly and completely to a spiritual pursuit (and I don't think it matters whether this be as a wandering ascetic or a householder) is not in the least a way of "dodging" the world, but of actually changing and addressing the world in the most authentic way possible.
In this sense, practicing yoga, or buddhism, or what have you IS the most revolutionary act possible. Just as Foucault often pointed out when people criticized him for not "doing" anything; thinking is doing and it is only our limited concept of what counts as "doing" which prevents of us from realizing this. So too with our spiritual lives.
Thank you for this wonderful piece.
Hey Thaddeus, point well taken. Bob, thanks for the props! BTW: I edited it since you read it, and I think it's much improved. Carol! Always great to hear from you. I contributed to your book project and look forward to reading your material there.
[…] […]
[…] As we move out of denial, we will become more physically healthy and vibrant. We will develop greater mental and emotional capacity to deal with reality as it is. […]
[…] and a refusal to be a grown up. So, as a handy guide, I have compiled a list of new agey, sensitive types that my friends and I have learned to avoid like the […]


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PALM BEACH, Fla. — Collections of 1920s Rene Lalique glass, netsuke carving and signed estate jewelry will headline a 500-lot estate auction planned for Oct. 27.
The auction, hosted by A.B. Levy's, will get underway at 1 p.m. It will also feature highly collectible timepieces by Patek Philippe, 19th- and 20th- century furniture, sterling silver and works of fine art.
Dubbed "Magnificent Jewels, Art, Antiques and Asian Works of Art," the auction will feature merchandise from prominent area estates and collections.
"I've never seen a collection of Netsuke quite as exciting as the one we have in this sale," said Albert Levy of A.B. Levy's.
An example lot is a Meiji period (1868-1912) Japanese carved wood-and-lacquer netsuke of a suit of armor atop a storage trunk. Measuring just two inches in height, the piece carries a pre-sale estimate of $500 to $800.
In addition to netsuke, the Asian category will feature jade, coral, snuff bottles, ivory, lapis, tiger's eye and a few porcelain pieces. Highlight lots will include a Qing Dynasty Chinese carved red-coral group depicting two beauties, each holding an attribute, mounted on a carved ivory base, which is estimated at $40,000 $60,000; and a Chinese carved jade figure of a beauty, shown standing wearing flowing robes, with an estimate of $10,000 to $20,000.
Nearly 75 lots of Rene Lalique glass, all from circa 1920s, will be offered in consecutive lots. Featured items include a molded opalescent glass statuette titled "Suzanne," signed Rene Lalique. The model, first introduced in 1925, is expected to command between $20,000 and $30,000.
Top lots in jewelry include an important sapphire and diamond ring, centered by an octagonal-shaped brilliant cut sapphire certified to be from Sri Lanka. Weighing 15.02 carats, the center stone is flanked by triangle-shaped diamonds. The piece carries an estimated price of $12,000 to $18,000.
Many jewelry lots signed by David Webb will also be offered. One is an 18kt platinum-and-diamond rock-crystal ring, centered by an emerald-cut diamond weighing approximately 3.25 carats, with an estimated price of $20,000 to $30,000. There are additional diamonds on each side weighing about one carat.
Certain to attract keen bidder interest is a fine collection of Patek Philippe timepieces. An example is an 18kt white-gold perpetual calendar chronograph with a mechanical movement with moon phases, AM/PM indicator and leap year indicator, which holds an estimate of $70,000 to $100,000. The watch comes with a sapphire case back, the original box and all the original paperwork dating to 2006.
From the same group is a Patek Philippe 18kt yellow-gold keyless pocket watch designed in the style of Louis XV and made circa 1890, with an estimate of $15,000 to $30,000. The domed back cover is decorated with a scene of St. George and the Dragon and the interior of the case has a scene of Christ on a ship in a rough sea. It has radial blue Roman numerals on oval white enamel reserves.
In addition to a date-of-sale morning preview, daily previews will be held Oct. 21 through Oct. 26, from 10 a.m. to 9 p.m. For those unable to attend in person, Internet bidding will be available via LiveAuctioneers.com, Artfact.com and The-Saleroom.com. Phone and left bids will also be taken.
For more information and to view the sale catalogue, visit A.B. Levy's website.
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Seared Scallops with Browned Butter Lemon Sauce
I'll never ever understand people who say they dislike chocolate or who say they dislike seafood. Those are some of my all time favorite foods (not together though… ewh). I love fish and shellfish but one of my favorite spoils of the sea is definitely sea scallops. They're pretty much perfect in my book. Quick to cook and when done right they are slightly sweet, super tender, with a golden brown mouthwatering crust. The only problem is that they can be pretty pricey. To my sheer joy giant sea scallops were on sale at my grocery store last week so I just had to make this recipe. This recipe is pretty foolproof too so you won't have to worry much about ruining your expensive scallops.
Recipe Rundown
Taste: Nutty, earthy, sweet, tart, and fresh. All around savory goodness.
Texture: This is the best part of seared scallops! They're crisp and crusty at the edges and fall-apart tender in the center. I loooove scallops.
Ease: Surprisingly easy but impressively gourmet and tasty.
Appearance: Just b-e-a-u-tiful.
Pros: Quick, easy, amazingly scrumptious.
Cons: None.
Would I make this again? Yes.
Seared Scallops with Browned Butter Lemon Sauce
Yield: 4 servings
For the sauce:
4 tablespoons unsalted butter, cut into 4 pieces
1 small shallot, minced
1 tablespoon minced fresh parsley
1/2 teaspoon minced fresh thyme
2 teaspoons lemon juice
Salt and pepper
For the scallops:
1 1/2 pounds large sea scallops, tendons removed and patted very dry
Salt and pepper
2 tablespoons vegetable oil
2 tablespoons unsalted butter
For the sauce:
Heat the butter in a small saucepan over medium heat and cook, swirling constantly, until the butter turns a dark golden brown color and has a nutty aroma, about 4 to 5 minutes. Be careful not to burn. Add the shallot and cook until fragrant, about 30 seconds. Remove the pan from heat and stir in the parsley, thyme, and lemon juice. Season to taste with salt and pepper. Cover the saucepan with a lid to keep warm.
For the scallops:
Sprinkle the dry scallops with salt and pepper on both sides. Heat 1 tablespoon of the oil in a 12-inch nonstick skillet over high heat until just smoking. Add half of the scallops in a single layer, flat side down, and cook, without moving, until well browned, 1 to 2 minutes.
Add 1 tablespoon of the butter to the skillet. Using tongs, carefully flip the scallops and continue to cook until the sides of the scallops are firm and the centers are opaque, 30 to 90 seconds. Remove the scallops to a plate and tent loosely with foil. Wipe out the skillet and repeat cooking with the remaining oil, scallops, and butter. Serve immediately with browned butter sauce.