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Logo of nihpaAbout Author manuscriptsSubmit a manuscriptHHS Public Access; Author Manuscript; Accepted for publication in peer reviewed journal;
Nature. Author manuscript; available in PMC 2011 September 24.
Published in final edited form as:
PMCID: PMC3109908

Comprehensive analysis of the chromatin landscape in Drosophila

Chromatin is composed of DNA and a variety of modified histones and non-histone proteins, which impact cell differentiation, gene regulation and other key cellular processes. We present a genome-wide chromatin landscape for Drosophila melanogaster based on 18 histone modifications, summarized by 9 prevalent combinatorial patterns. Integrative analysis with other data (non-histone chromatin proteins, DNaseI hypersensitivity, GRO-seq reads produced by engaged polymerase, short/long RNA products) reveals discrete characteristics of chromosomes, genes, regulatory elements, and other functional domains. We find that active genes display distinct chromatin signatures that are correlated with disparate gene lengths, exon patterns, regulatory functions, and genomic contexts. We also demonstrate a diversity of signatures among Polycomb targets that include a subset with paused polymerase. This systematic profiling and integrative analysis of chromatin signatures provides insights into how genomic elements are regulated, and will serve as a resource for future experimental investigations of genome structure and function.

The model organism Encyclopedia of DNA Elements (modENCODE) project is generating a comprehensive map of chromatin components, transcription factors, transcripts, small RNAs, and origins of replication in D. melanogaster and C. elegans1,2. Drosophila has been used as a model system for over a century to study chromosome structure and function, gene regulation, development, and evolution. The availability of high-quality euchromatic and heterochromatic sequence assemblies3-5, extensive annotation of functional elements6, and a vast repertoire of experimental manipulations enhance the value of epigenomic studies in Drosophila.

Genome-wide profiling of chromatin components provides a rich annotation of the potential functions of the underlying DNA sequences. Previous work has identified patterns of post-translational histone modifications and non-histone proteins associated with specific elements (e.g. transcription start sites, enhancers), as well as delineating the transcriptional status of genes and large domains7,8. Here, we present a comprehensive picture of the chromatin landscape in a model eukaryotic genome. We define combinatorial chromatin ‘states’ at different levels of organization, from individual regulatory units to the chromosome level, and relate individual states to genome functions.

Combinatorial chromatin states

We performed chromatin immunoprecipitation (ChIP)-array analysis for numerous histone modifications and chromosomal proteins (Supp. Table 1), using antibodies tested for specificity and cross-reactivity9 (Supp. Figure 1). Here, we describe analyses of cell lines S2-DRSC (S2) and ML-DmBG3-c2 (BG3), derived from late male embryonic tissues (stages 16-17) and the central nervous system of male third instar larvae, respectively (see for data from other cell lines and animal stages). Analysis reveals groups of correlated features, including those associated with heterochromatic regions10, Polycomb-mediated repression11, and active transcription12 (Supp. Figure 2), similar to those observed in other organisms13,14. This suggests that specific histone modifications work together to achieve distinct chromatin “states”.

We utilized a machine-learning approach to identify the prevalent combinatorial patterns of 18 histone modifications, capturing the overall complexity of chromatin profiles observed in S2 and BG3 genomes with 9 combinatorial states (Figure 1a, Methods). The model associates each genomic location with a particular state, generating a chromatin-centric annotation of the genome (Figure 1b). We examined each state for enrichment in non-histone proteins (Figure 1a, Supp. Figure 3) and gene elements, as well as distribution across the karyotype (Figure 1b, Supp. Figure 4) and finer-scale levels (Figure 1c-e).

Figure 1
Chromatin annotation of the Drosophila melanogaster genome

Most distinct chromatin states are associated with transcriptionally active genes. Active promoter and transcription start site (TSS)-proximal regions are identified by state 1 (Figure 1; red), marked by prominent enrichment in H3K4me3/me2 and H3K9ac. The transcriptional elongation signature associated with H3K36me3 enrichment is captured by state 2 (purple), found preferentially over exonic regions of transcribed genes. State 3 (brown), typically found within intronic regions, is distinguished by high enrichment in H3K27ac, H3K4me1, and H3K18ac. A related chromatin signature is captured by state 4 (coral), distinguished by enrichment of H3K36me1, but notably lacking H3K27ac. The number of genes associated with each chromatin state and the distribution of states within genes are shown in Supp. Figure 5.

Several aspects of large-scale organization are revealed by the karyotype view (Figure 1b). Chromosome X is strikingly enriched for state 5 (green), distinguished by high levels of H4K16ac in combination with some enrichment in H3K36me3 and other marks of “elongation” state 2 (a pattern associated with dosage compensation in male cells15). Pericentromeric heterochromatin domains and chromosome 4 are characterized by high levels of H3K9me2/me3 (state 7, dark blue)10. Finally, the model distinguishes another set of heterochromatin-like regions containing moderate levels of H3K9me2/me3 (state 8, light blue, Figure 1e). Surprisingly, this state occupies extensive domains in autosomal euchromatic arms in BG3 cells, and in chromosome X in both cell lines16.

Further aspects of chromatin organization can be visualized by folding the chromosome using a Hilbert curve (Figure 2a)17, which maintains the spatial proximity of nearby elements. Thus, local patches of corresponding colors reveal the sizes and relative positions of domains associated with particular chromatin states (Figure 2b; Supp. Figures 6-9). For instance, specks of TSS-proximal regions (state 1) are typically contained within larger blocks of transcriptional elongation marks (state 2), which are in turn encompassed by extensive patches of H3K36me1-enriched domains (state 4) and variable-sized blocks of state 3. The clusters of open chromatin formed by these gene-centric patterns are separated by extensive silent domains (state 9) and regions of Polycomb-mediated repression (state 6). Factors responsible for domain boundaries were not identified in our analysis (Supp. Figure 10).

Figure 2
Visualization of spatial scales and organization using compact folding

We also developed a multi-scale method to characterize chromatin organization at the spatial scale appropriate for the genome properties being investigated. For example, we observe that chromatin patterns most accurately reflect the replication timing of the S2 genome at scales of ~170kb (Supp. Section 1). This is consistent with size estimates of chromatin domains influencing replication timing18, and suggests that multiple replication origins are coordinately regulated by the local chromatin environment (each replicon is ~28-50kb 19).

To examine combinatorial patterns not distinguished by the simplified 9-state model, we also generated a 30-state combinatorial model that utilizes presence/absence probabilities of individual marks20 (Supp. Figure 11). The increased number of states may identify finer variations that are biologically significant, e.g., a signature corresponding to transcriptional elongation in heterochromatic regions16.

Chromatin state variation among genes

Active genes generally display enrichments or depletions of individual marks at specific gene segments (Figure 3a). When classified according to their chromatin signatures (Supp. Figure 12), active genes fall into subclasses correlated with expression magnitude (Supp. Section 2), gene structure, and genomic context (e.g. heterochromatic genes combine H3K9me2/me3 with some active marks). Of particular interest is one class of long expressed genes, many with regulatory functions, which are enriched for H3K36me1 (cluster 2, Supp. Figure 12; 131 genes in S2, 202 in BG3; Supp. Table 2).

Figure 3
Chromatin patterns associated with transcriptionally active genes

To further examine the patterns associated with long genes, we clustered expressed autosomal genes ≥4kb based on blocks of enrichment for each chromatin mark (Figure 3b; 1055 genes). We observe that genes with large 5’-end introns (green subtree, Figure 3b; 552 genes) show extensive H3K27ac and H3K18ac enrichment, broader H3K9ac domains, and blocks of H3K36me1 enrichment (chromatin state 3, Figure 3b, last column). These genes are enriched for developmental and regulatory functions (Supp. Table 3), and are positioned within domains of Nipped-B21 (Figure 3b), a cohesin-complex loading protein previously associated with transcriptionally active regions21,22. In contrast, genes with more uniformly distributed coding regions (red subtree, Figure 3b) lack most state 3 marks, and H3K9ac enrichment is restricted to the 2kb downstream of the TSS. These differences are not explained by variation in histone density (Supp. Figure 13). Overall, the presence or absence of state 3 is the most common difference in the chromatin composition of expressed genes that are 1kb and longer (Supp. Figure 14), and the presence of state 3 consistently correlates with a reduced fraction of coding sequence in the gene body, mainly associated with the presence of a long first intron.

State 3 domains are highly enriched for specific chromatin remodeling factors (SPT16 and dMI-2; Supp. Figures 15,16), whereas state 1 regions around active TSSs are preferentially bound by NURF301 and MRG15. ISWI is enriched in both states 1 and 3 (Supp. Figures 16,17). State 3 domains also exhibit the highest levels of nucleosome turnover23, and show higher enrichment of the transcription-associated H3.3 histone variant24 than either the TSS- or elongation-associated states 1 and 2 (Supp. Figures 15,16). Consistent with earlier analyses of cohesin-bound regions25, state 3 sequences tend to replicate early in G1 phase, and show abundance of early replicating origins (Supp. Figure 18). A regulatory role for state 3 domains is suggested by enrichment for a known enhancer binding protein (dCBP/p300 26) in adult flies, and for enhancers validated in transgene constructs27 (Supp Figure 19).

Modes of regulation in Polycomb domains

In Drosophila, loci repressed by Polycomb group (PcG) proteins are embedded in broad H3K27me3 domains that are regulated by Polycomb Response Elements (PREs) bound by E(Z), PSC, and dRING (Figure 1d)28,29. We find that regions of H3K4me1 enrichment surround all PREs, 90% of which also display narrower peaks of H3K4me2 enrichment (Supp. Figure 20). While this pattern is reminiscent of transcriptionally-active promoter regions, PREs lack H3K4me3, suggesting that a different mechanism of H3K4 methylation is employed, perhaps involving the Trithorax H3K4 histone methyltransferase (HMTase) found at all PREs29.

To examine chromatin states associated with PcG targets, we analyzed the chromatin and transcriptional signatures of TSSs in Polycomb-bound domains (Figure 4a, Supp. Figure 21). In addition to fully repressed TSSs (cluster 1, Figure 4a), we identify TSSs maintained in the “balanced” state29 (cluster 2, Figure 4a), distinguished by coexistence of Polycomb with active marks (including the HMTase ASH1) and production of full-length mRNA transcripts (e.g. Psc domain, Figure 1d).

Figure 4
Signatures of TSSs within domains of Polycomb-mediated repression

TSSs in clusters 3 and 4 are distinguished by the presence of adjacent PREs (Figure 4a). Surprisingly, 53% of the PRE-proximal TSSs produce short RNA transcripts30 (cluster 3, Figure 4a), suggesting stalling of engaged RNA pol II 30. Using the global run-on sequencing (GRO-seq) assay to accurately assess engaged RNA polymerases31, we observe that cluster 3 TSSs produce short transcripts in the sense orientation. The level of GRO+ signal is similar to that found at fully-transcribed genes (Supp. Figure 22); thus, transcription initiates in cluster 3, but elongation fails. Interestingly, these genes are enriched for regulatory and developmental functions, even more than other genes within Polycomb domains (see Supp Tables 4,5). Genes without TSS-proximal PREs generally lack short transcript signatures (e.g. clusters 1 in Figure 4a; see Supp. Figure 21 for exceptions). Importantly, engaged polymerases and transcripts are not a general feature of PREs; TSS-distal PREs typically lack short RNA and GRO-seq signals (Figure 4b, Supp. Figure 22) despite being similarly enriched in H3K4me1/me2. The striking link between TSS-proximal PREs and the production of short RNAs suggests a potential mechanism for control of these developmental regulatory genes, whereby the same features that recruit H3K4 methyl marks to PREs also facilitate RNA pol II recruitment to nearby TSSs.

DHS plasticity and chromatin states

We utilized a DNase I hypersensitivity assay32,33 to examine the distributions of putative regulatory regions and their relationships with chromatin states. DHS mapping broadly identifies sites with low nucleosome density and regions bound by non-histone proteins34,35. Short-read sequencing identified 8616 high-magnitude DNase I hypersensitive sites (DHSs) in S2 cells and 6354 in BG3 cells (and a comparable number of low-magnitude DHSs, Supp. Figure 23; see Methods). Approximately half of the high-magnitude DHSs are found at transcriptionally-active TSSs (Supp. Figure 24). Thus, the chromatin context of the TSS-proximal DHSs is dominated by the features expected for an active TSS, including RNA Pol II, H3K4me3 and other state 1 marks (clusters 1,2 Figure 5a, Supp. Figure 25).

Figure 5
Chromatin signatures of regulatory elements identified by DNaseI hypersensitivity

Of the 36% TSS-distal DHSs, most (60%) are positioned within annotated expressed genes (Supp. Figure 24). These gene-body DHSs are distinguished from TSS-proximal DHSs by low H3K4me3, higher levels of H3K4me1, H3K27ac, and other marks linked to chromatin state 3 (clusters 3,4 Figure 5a, Supp. Figure 26). An additional 20% of the TSS-distal DHSs are outside of annotated genes, but show signatures associated with active transcription starts or elongation, suggesting new alternative promoters or unannotated genes (Supp. Figures 27,28). The remaining 20% of TSS-distal DHSs that appear intergenic (6% of all DHSs) are typically enriched for H3K4me1, but lack other active marks (cluster 5, Figure 5a).

Most DHS positions fall into the TSS-proximal state 1 or the intron-biased state 3 (Figure 5b). State 3 lacks H3K4me3 and is enriched for H3K4me1/H3K27ac/H3K18ac, similar to mammalian enhancer elements36,37. Many state 3 DHS positions bind regulatory proteins: GAGA factor binds to 49% of these DHSs in S2 cells, and developmental transcription factors bind to 44% of these DHSs in embryos38. Intriguingly, we find that TSS-distal DHSs in Drosophila exhibit low-level bi-directional transcripts (Figure 5a shortRNA panel, Supp Figures 29,30), analogous to the enhancer RNAs (eRNAs) characterized in mice39. Analysis of GRO-seq data (Figure 5e) suggests that eRNA-like transcripts are common to both intra- and inter-genic TSS-distal DHSs in Drosophila, a feature that is conserved with mammals.

The association of DHSs with chromatin states 1 and 3 (Figure 5c) persists even in chromosome 4 and pericentromeric heterochromatin, where such states are infrequent (Supp. Figure 31). This suggests that these chromatin states and associated remodeling factors (e.g. ISWI, SPT16) provide the context necessary for non-histone chromosomal protein binding at DHSs, or are the consequence of such binding events. To investigate this interdependency, we analyzed a high-confidence set of loci that exhibit DHSs in only one of the two examined cell lines (Supp. Figure 32). Surprisingly, although in general more DHSs are in state 1 regions, 91% of the cell type-specific DHSs are found within state 3 domains (14-fold increase compared to state 1 DHSs; Supp. Table 6, Figure 5d). Comparison with DHSs in an additional cell type (Kc167, Supp. Figure 33) confirms that DHSs displaying plasticity between cell types are mostly found in state 3. When DHSs are absent, the altered loci maintain chromatin state 3 in 23% of the cases (Figure 5d), indicating that the presence of state 3 is not always dependent on the DHS. More frequently, the altered loci transition to state 4 (43% of the cases), an open chromatin state that lacks many of the histone modifications and chromatin remodelers characteristic of state 3. While the less frequent transitions to the Polycomb state 6 (7%) or background state 9 (17%) typically coincide with gene silencing, most of the genes that maintain state 3 or transition to state 4 remain transcriptionally active (Supp. Figure 34). These observations provide further support for an enhancer-like function for state 3 DHSs, and suggest a more subtle regulatory role than simple linkage to the presence or absence of gene expression.

Chromatin annotation of genome functions

The genomic chromatin state annotation and discovery of refined chromatin signatures for chromosomes, domains, and subsets of regulatory genes demonstrate the utility of a systematic, genome-wide profiling of an organism that is already understood in considerable detail. Clearly, the definition and functional annotation of chromatin patterns will be enhanced by incorporation of data for different types of components. Five ‘colors’ of chromatin were recently identified in Kc167 cells using chromosomal protein maps40. Comparison with our 9-state model shows similarities as well as differences in the ability to distinguish functional elements (Supp. Figure 35); thus, further integration of such data in the same cell type may resolve additional functional features. Our results illustrate the utility of integrating multiple data types (histone marks, non-histone proteins, chromatin accessibility, short RNAs, and transcriptional activity) for comprehensive characterization of functional chromatin states.

An important, repeated theme is that chromatin state analysis identifies unexpected distinctions between subsets of active genes. Besides the differences linked to genomic context (e.g., male X chromosome, heterochromatin), the main source of variability is the presence of the acetylation-rich state 3 (Figure 6). Several lines of evidence suggest that the intronic positions marked by state 3 are important for gene regulation. State 3 regions show specific associations with known chromatin remodelers (SPT16, dMi-2 and ISWI) and gene regulatory proteins (e.g. GAF, dCBP/p300), and the highest rates of nucleosome turnover and transcription-dependent deposition of the H3.3 variant. State 3 genes are also bound by cohesin complex proteins, thought to associate with decondensed chromatin21 to promote looping interactions with promoter regions22.

Figure 6
Spatial arrangements of chromatin states associated with active transcription

A regulatory role for state 3 chromatin is further suggested by the high density of DHSs, comparable to that of active TSS state 1, and the fact that state 3 accounts for most of the DHS plasticity among cell types. The combinations of histone marks found in state 3 are similar to signatures of mammalian enhancers36, which also show high variability between cell types37. Furthermore, state 3 DHSs exhibit low levels of short, non-coding bidirectional transcripts reminiscent of eRNAs identified in mice39. Together, these findings suggest that state 3 regions contain enhancers or other regulatory elements, and that a combination of modifications can be used to identify new elements in the genome.

Genes within repressive Polycomb domains also display several distinct combinatorial chromatin patterns (Figure 4a), which likely represent a range of functional states: repressed, paused, or expressed genes in either balanced29 or fully activated states. Alternatively, distinct signatures might mark subsets of regulatory genes that require either long-term repression or the ability to reverse functional states, depending on environmental or developmental cues. The PRE-proximal paused TSSs have some similarity to the “bivalent” genes in mammalian cells, which also display transcriptional pausing of key regulatory and developmental genes41,42. However, the mammalian “bivalent state” is characterized by the simultaneous presence of PcG proteins, H3K27me3 and H3K4me3, which in Drosophila is found only in the fully-elongating “balanced” state29,43.

In summary, comprehensive analysis of chromatin signatures has enormous potential for annotating functional elements in both well-studied and new genomes. Going forward, our systematic characterization of the epigenomic and transcriptional properties of Drosophila cells should spur in-depth experimental analyses of the relationship between chromatin states and genome functions, ranging from whole chromosomes down to individual regulatory elements and circuits.

Methods Summary

Histone modification and chromosomal protein antibodies were characterized for cross-reactivity. ChIP-chip was performed in duplicate, using Affymetrix Drosophila Tiling 2.0R Arrays. Digital DNaseI-seq assays were performed as described previously44, and Global Run-On library (GRO-seq) data was generated as described in Core et al31. Short RNA data was generated by Nechaev et al30, and RNA-seq data was generated by Graveley et al.45. The chromatin state models were generated as Hidden Markov Models of different histone marks. DHSs were identified as read density peaks significantly enriched relative to the genomic DNA control. Clustering of chromatin signatures was determined using the PAM algorithm.

Growth conditions

ML-DmBG3-c2 cells were obtained from DGRC (, and S2-DRSC cells were from the DRSC ( All cell lines were grown to a density of ~5×106 cells/ml in Schneider’s media (Gibco) supplemented with 10% FCS (HyClone). 10 μg/ml insulin was added to the ML-DmBG3-c2 media.

Antibodies are listed in Supplemental Table 1. Commercial antibodies against modified histones were tested by Western-blot for the lack of cross-reactivity with the corresponding recombinant histone produced in E.coli and non-histone proteins from embryonic nuclear extracts. Antibody specificity was further assayed by Western dot/slot blot against a panel of synthetic modified histone peptides. Only antibodies that showed <50% of total signal associated with non-histone proteins, and more than 5-fold higher affinity for the corresponding histone peptide, were used in ChIP experiments.

The specificity of antibodies against chromosomal proteins was tested by Western blots with nuclear extracts prepared from mutant flies or S2 cells subjected to RNAi knockdown.46. An antibody was considered specific if it recognized a major band of expected mobility that was absent in the sample prepared from mutant flies, or diminished 2-fold or more after RNAi depletion. When possible, distributions of a chromosomal protein were mapped with two antibodies generated against different epitopes (see Supp. Figure 17). Data from chromatin proteins for which only one antibody was available was validated by comparison with published genomic distributions for a different component of the same complex, or to published genomic distributions generated with a different antibody.

ChIP and microarray hybridization

Crosslinked chromatin from cultured cells was prepared as described in Schwartz et al.28 with the following modifications. Prior to ultrasound shearing, cells were permeabilized with 1% SDS, and shearing was done in TE-PMSF (0.1% SDS, 10mM Tris-HCl pH8.0, 1mM EDTA pH8.0, 1mM PMSF) using a Bioruptor (Diagenode) (2 × 10 min, 1 × 5 min; 30sec on, 30 sec off; high power setting).

ChIP was performed as in Schwartz et al.28 and IP’d DNA was amplified using the whole genome amplification kit (WGA2, Sigma) according to the manufacturer’s instructions (chemical fragmentation step was omitted). The amplified material was labeled and hybridized to Drosophila Tiling Arrays v2.0 (Affymetrix) as in Schwartz et al.28.

Processing of ChIP data

At least two independent biological replicates were assessed for each ChIP profile. The log2 intensity ratios (M values) were calculated for each replicate. The profiles were smoothed using local regression (lowess) with 500bp bandwidth, and the genome-wide mean was subtracted. The regions of significant enrichment were determined as clusters of at least 1kb in length, with gaps no more than 100bp where M value exceeds a statistically significant (0.1% FDR) enrichment threshold. The set of biological replicates was deemed consistent if the enriched regions from individual experiments had a 75% reciprocal overlap, or if at least 80% of the top 40% of the regions identified in each experiment were identified in the other replicate (before comparison the replicates were size-equalized by increasing the significance threshold for a replicate with more enriched sequence). The data from individual replicates were then combined using local regression smoothing, and used for all of the presented analysis, unless noted otherwise.

DNaseI hypersensitivity

Digital DNaseI-seq assays were performed as described previously44. The sequenced reads were aligned to the dm3 genome assembly, recording only uniquely mappable reads. To detect DNase I hypersensitive sites, hotspot positions were identified based on a 300bp scanning window statistic (Poisson model relative to 50kb background density, Z-score threshold of 2), and peaks of read density were selected within the hotspots using randomization-based thresholding at 0.1% FDR. The set of high-magnitude DHSs analyzed here (except for Supp. Figure 23) was identified as a subset of all peaks that show statistically significant enrichment over the normalized genomic DNA read density profile (using a 300bp window centered around the peak, binomial model, with Z-score threshold of 3). This method controls for copy number variation and sequencing/mapping biases, however it may also reduce the sensitivity of DHS detection. In the DHS chromatin profile clustering analysis (Figure 5a, relevant supplementary figures), DHSs found within 1kb of another DHS were excluded if their enrichment magnitude (relative to genomic background) was lower (to avoid showing the same region more than once).

RNA sequencing

The preparation of RNA-seq libraries and sequencing is described in Graveley et al.45. The sequenced reads were aligned to the dm3 genome assembly and annotated exon junctions, recording only uniquely mappable reads. The RPKM (reads per kilobase of exonic sequence per million reads mapped) was estimated for each exon. The total transcriptional output of each annotated gene was estimated based on the maximum of all exons within the gene. The presented analysis uses log10(RPKM+1) values unless otherwise noted.

GRO sequencing

Global Run-On library was prepared from S2 cells and sequenced as described in Core et al31. The reads were aligned to the dm3 genome assembly, recording only uniquely mappable reads. The smoothed profiles of reads mapping to each strand were calculated using Gaussian smoothing (σ=100bp). The analysis uses log10(d+1), where d is the smoothed density value.

Short RNA data processing

The short RNA data for S2 cells was generated by Nechaev et al30, and was aligned and processed in the same way as the GRO-seq data.

Chromatin state models

To derive a nine-state joint chromatin state model for S2 and BG3 cells (Figure 1a), the genome was first divided into 200bp bins, and the average enrichment level was calculated within each bin based on unsmoothed log2 intensity ratio values taking into account individual replicates, using all histone enrichment profiles and PC to discount the genome-wide difference in S2 H3K27me3 profiles. The bin-average values of each mark were shifted by the genome-wide mean, scaled by the genome-wide variance, and quantile-normalized between the two cells. The HMM with multivariate normal emission distributions was then determined from the Baum-Welch algorithm using data from both cell types, and 30 seeding configurations determined with K-means clustering. States with minor intensity variations (Euclidian distance of mean emission values < 0.15) were merged. Larger models (up to 30 states) were examined, and the final number of states was chosen for optimal interpretability.

An extensive discrete chromatin state model (Supp. Figure 11) was calculated as described in Ernst et al.20. The model was trained using 200bp grid with binary calls (enriched/not enriched). The binary calls were made based on a 5% FDR threshold determined from 10 genome-wide randomizations for each mark. For H1, H4 and H3K23ac regions of significant depletion rather than enrichment were called.

Regions of enrichment for individual marks (Figure 3)

To determine contiguous regions of enrichment for individual marks, a three-state HMM was used, with states corresponding to enriched, neutral, and depleted profiles (normally-distributed emission parameters: (μ=[-0.5 0 0.5], σ2=0.3). The enriched regions were determined from the Viterbi path. The HMM segmentation was applied to unsmoothed M value data taking into account individual biological replicates. The genes were clustered based on the combinatorial pattern of occurrence of enriched regions (coding exons and state panels were not used for clustering).

Classification of enrichment profiles (Figures 4,,55)

Clustering of chromatin signatures around TSSs (Figure 4a), PREs (Figure 4b), and DHSs (Figure 5a, relevant supplements) was determined using the PAM algorithm. For clustering, each profile was summarized with average values within bins spanning ±2kb regions. 100bp bins were used for the central ±500bp region, 300bp bins outside.

Supplementary Material

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We thank our technicians David Acevedo, Sarah Gadel, Cameron Kennedy, Ok-Kyung Lee, and Sarah Marchetti and Rutgers BRTC. We also thank our colleagues who donated antibodies: J. Kadonaga (H1), A.L. Greenleaf (RNA pol II), G. Reuter (SU(VAR)3-9), G. Cavalli (GAF), and I.F. Zhimulev/H. Saumweber (Chromator). The major support for this work came from the modENCODE grant U01HG004258 to G.H.K (PI) and S.C.R.E., M.I.K., P.J.P., and V.P. (co-PIs). Additional funding came from RC2 HG005639, U01 HG004279, R01 GM082798, R37 GM45744, RC1 HG005334, and NSF 0905968.

Author contribution P.V.K. performed most bioinformatic analysis. A.A.A., Y.B.S., A.M., N.C.R., E.L., A.A.G., T.G., D.L., A.P., and G.S. generated data, directed by S.C.R.E., M.I.K., V.P., and G.H.K. The 30-state analysis was performed by J.E., and M.K., while M.Y.T., L.J.L., R.X., Y.L.J., R.P., and E.P.B. performed additional bioinformatic analysis. P.J.S., T.P.C., R.S., R.E.T., and J.A.S. generated and processed DHS data. D.M.M. helped with replication analysis. P.J.P. supervised all analysis. G.H.K. coordinated the entire project. P.V.K., G.H.K., and P.J.P. wrote the manuscript, with contributions from S.C.R.E., M.I.K., V.P., Y.B.S, N.C.R, A.A.A, and A.M.

Author information The data are available from modENCODE site: GRO-seq data is available from Gene Expression Omnibus (GEO GSE25321). The authors declare no competing financial interests.

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26. Visel A, et al. ChIP-seq accurately predicts tissue-specific activity of enhancers. Nature. 2009;457:854–858. [PMC free article] [PubMed]
27. Zinzen RP, Girardot C, Gagneur J, Braun M, Furlong EE. Combinatorial binding predicts spatio-temporal cis-regulatory activity. Nature. 2009;462:65–70. [PubMed]
28. Schwartz YB, et al. Genome-wide analysis of Polycomb targets in Drosophila melanogaster. Nat Genet. 2006;38:700–705. [PubMed]
29. Schwartz YB, et al. Alternative Epigenetic Chromatin States of Polycomb Target Genes. PLoS Genet. 2010;6:e1000805. [PMC free article] [PubMed]
30. Nechaev S, et al. Global analysis of short RNAs reveals widespread promoter-proximal stalling and arrest of Pol II in Drosophila. Science. 2010;327:335–338. [PMC free article] [PubMed]
31. Core LJ, Waterfall JJ, Lis JT. Nascent RNA sequencing reveals widespread pausing and divergent initiation at human promoters. Science. 2008;322:1845–1848. [PMC free article] [PubMed]
32. Wu C. The 5’ ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature. 1980;286:854–860. [PubMed]
33. Wu C, Bingham PM, Livak KJ, Holmgren R, Elgin SC. The chromatin structure of specific genes: I. Evidence for higher order domains of defined DNA sequence. Cell. 1979;16:797–806. [PubMed]
34. Elgin SC. The formation and function of DNase I hypersensitive sites in the process of gene activation. J Biol Chem. 1988;263:19259–19262. [PubMed]
35. Jin C, et al. H3.3/H2A.Z double variant-containing nucleosomes mark ‘nucleosome-free regions’ of active promoters and other regulatory regions. Nat Genet. 2009;41:941–945. [PMC free article] [PubMed]
36. Heintzman ND, et al. Distinct and predictive chromatin signatures of transcriptional promoters and enhancers in the human genome. Nat Genet. 2007;39:311–318. [PubMed]
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38. MacArthur S, et al. Developmental roles of 21 Drosophila transcription factors are determined by quantitative differences in binding to an overlapping set of thousands of genomic regions. Genome Biol. 2009;10:R80. [PMC free article] [PubMed]
39. Kim TK, et al. Widespread transcription at neuronal activity-regulated enhancers. Nature. 2010;465:182–187. [PMC free article] [PubMed]
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Monday, December 7, 2009

Christmas Crab Tree

For those of you who don't know, our last name is Crabtree. A few years ago, we created a family tradition of collecting crab ornaments for our Christmas "Crab Tree". Friends & family have given us wonderful crab ornaments they have found for gifts during the holiday. And each year, we make a new crab ornament to add to our tree. This truly is my boys favorite decoration. While they decorate their tree, they talk about each crab ornament & the special people who helped us create our Christmas Crab Tree.

This years crab ornament creation.....

What is your special holiday tradition that you enjoy with your family?
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Microsoft Expects Windows Phone 7 Manufacturing Costs To Drop More Than Half Next Year

Microsoft Corporation (NASDAQ:MSFT) expects the production costs of handsets that uses Windows Phone 7 software to drop by over half next year. Qualcomm Inc. will allow phones that run on the Windows Phone 7 operating system to be produced for under $200. This is a substantial drop from the $400 manufacturing cost from when the mobile operating system was introduced last year. This announcement was made by Windows Phone head Andy Lees. Microsoft works exclusively with Qualcomm for chip development. [BusinessWeek]

This article was written by Amit Chowdhry. You can follow me at @amitchowdhry or on Google+ at
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Monday, July 16

Thank you, Riehl World View

Dan Riehl at Riehl World View linked to my post today (I don't care that Egyptians threw tomatoes at Hillary Clinton) and delivered this on-target introduction:
What's a few tomatoes, when Americans are being killed?

Unlike our mainstream media, Pundita seems a bit more upset over Americans being killed, than a couple tomatoes hitting a car. Is that so wrong?
Dan is the kind of person I'd want at my side in a trench. What am I saying? Going up against Washington on the matter of its support for Pakistan's military is already a form of trench warfare. Thanks, Dan.

Yes, why does this not upset people more?

Our institutional ideas about strategy mean that institutions view fealty to a certain national strategy as overcoming these concerns.

But that is obscene when not in an actual existential war.
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Monday, November 16, 2009

Conservatives Sit Down For Canada and Leave National Portrait Gallery Homeless

Paul Dewar of the NDP worked very hard to get the Reform Conservatives to stick to Jean Chretien's plan to keep the National Portrait Gallery in the Nation's capital. Instead after flitting about, they gave the contract to Calgary. Calgary? A NATIONAL gallery in Calgary? Why were we not surprised?

However, as it turned out, the whole fiasco was a lesson in futility, wrought with lies; and we are no further ahead than we were when the campaign first started. Our gallery remains homeless and Conservative incompetence has hurt taxpayers again. When are they going to start taking an interest in Canadian culture? But then Harper doesn't believe Canada has a culture but is simply part of a larger Anglo culture, so what hope is there? Our treasures remain in a warehouse until we can elect a government that gives a damn.

Portrait gallery panel chafes at choice restrictions: documents
By Dean Beeby (CP)
November 15, 2009

OTTAWA — Members of a blue-ribbon panel created to find a home for the Portrait Gallery of Canada felt handcuffed because the federal government forbade them from examining key cost and tourism issues, says a newly released report.

The five-member panel was also misinformed at its crucial selection meeting that all five of the private-sector bids it was evaluating fully complied with federal rules - learning only months later that two did not.

The turmoil behind closed doors may help explain why the highly politicized search for a private-sector home for the portrait collection ended in a shambles a year ago, with no winning bid and the government abandoning the idea.

The Canadian Press obtained formerly secret accounts of deliberations, through requests under the Access to Information Act, after federal officials reneged on a promise to proactively release the materials.

In early 2007, the Conservative government ditched a plan by Jean Chretien's Liberal government to display the venerable portrait collection in the abandoned American embassy building on Wellington Street, facing Parliament Hill.

About $11.4 million had been spent on the project by that point, though some funds were used to remove asbestos and carry out other renovations that were needed no matter what the future use of the building. Total costs of the new gallery facility were projected in 2005 at $44.6 million.

The Tories instead announced that nine Canadian cities were eligible as a potential homes for the collection and invited private-sector bids, to encourage a public-private partnership and possibly place a cultural institution outside Ottawa.

A panel of five experts, including Canada's chief librarian and archivist Ian Wilson, was struck to evaluate bids and make recommendations. Initial expressions of interest came from 132 firms, including some in Halifax, Quebec City, Montreal and Toronto.

The deadline for submissions was changed twice, after federal officials failed to provide timely answers to basic questions from interested parties.

By May 16 last year, five formal proposals had arrived along with their $1-million deposits - from developers in Ottawa, Winnipeg, Calgary and two in Edmonton.

But panel members soon chafed when told they could not consider the costs of operating a new gallery at a distance from the storage facility in nearby Gatineau, Que., where the main collection resides.

A 2006 internal Canadian Heritage memo estimated the remote-location costs would add about $2.5 million to the $5.5-million annual operating bill of any new gallery located far from Ottawa. But this financial information was deemed non-relevant to the bid evaluations.

Panel members were also told they could not consider whether any of the five bids had strong potential for tourism growth.

Members expressed "dissatisfaction with some of the constraints," says a report by Jim Lynes, an independent consultant hired by government to monitor the process.

"There was no way to favour cities with a better potential to generate visitors for the PGC (Portrait Gallery of Canada)," says a censored version of the document.

"There was no opportunity to take the costs of a decentralized facility into consideration."

Federal officials told the panel those "constraints" could not be changed because they reflected "policy direction" from government.

A spokeswoman for Library and Archives Canada, which manages the portrait gallery, said in an interview that tourism had already been considered in creating the nine-city short list. Isabelle Vigneault also said any remaining cost issues would have been dealt with in a "second phase" of the bidding process.

Panel members held an intense selection meeting over two days in July 2008, complaining that some vital information had been provided only at the last minute. They were also given erroneous information on scoring, which cause confusion.

"Members were also concerned that they were not being given enough time to read the five submissions," says Lynes' report, which notes that all bids were officially deemed compliant with the requirements of the tender.

The panel eventually endorsed one bid (the identity is censored in the released material) on condition the government ask the developer to drop some provisions that tilted the deal too far in favour of the private-sector.

The provisions were "unacceptable in that they limited the Crown's rights and ability to act and increased the level of risk for the Crown, while at the same time limiting the risks for the developer."

Lynes' report of the deliberations added that some developers "seemed to fail to recognize that the Portrait Gallery of Canada is a national institution" - the very fear raised by some critics of the process.

The process unravelled in early November when federal lawyers deemed two of the bids - including the favoured one - non-compliant, contrary to assurances the panel was given more than three months earlier.

Lynes' final report, dated Dec. 15, 2008, concluded the bid-and-evaluation process that ultimately failed to produce a winner was technically fair and open, though had only "limited" transparency.

"Although transparency has, to this point, been limited, the planned release of the reports of the evaluation committee and the IRM (independent review monitor) will ensure that the process has been transparent," he wrote.

But the government appears to have had no such plan to proactively release the reports.

Internal Canadian Heritage media lines say the reports "will be made available through access to information," a formal procedure available to anyone wanting to access any government document.

The Canadian Press obtained heavily redacted versions of both reports through the Access to Information Act after months-long delays.

Lynes declined an interview, citing confidentiality agreements. Ian Wilson, who has since retired as librarian and archivist, also declined immediate comment.

"I continue to be bound by the post-employment code and my obligations as a former senior official," he said in an email.

The aborted bid-evaluation process cost taxpayers about $1.9 million, according to the documents.

The Portrait Gallery of Canada remains without a home, residing in the Gatineau warehouse, on a federal website and in occasional exhibitions around the country. The federal government has budgeted $3.5 million annually for the program.

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Watch City Of Fathers (2009) | Putlocker Movies HD

City Of Fathers (2009)

posted by admin, July 7, 2015 @ 11:18 pm

Movie: City Of Fathers

  • Director:
  • Stars:
  • Release Date: October 15, 2009
  • Run Time: 101 mins
  • Genre: Action, Drama, Korean

An action drama by Park Ji-won-I about a gambling street thug, Kang-su, who is always on the run from loan sharks. But when his young son Jong-chul is diagnosed with kidney cancer, he tries to be a real parent for the first time and seeks out Jong-chuls biological father, Tae-suk. Tae-suk, however, refuses to help. What will become of Jong-chul?

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Headlight Washers Not Working

Bodywork & Interior problems

Headlight Washers Not Working

Postby niallfagan » Tue Apr 05, 2016 12:09 am

My Headlamp washers aren't working on both sides. I have been told that they should work after ever 5 times i activate the windscreen washers, also that the bottle needs to be more than 50% full for headlight washers to work. So far i am having no joy. I asked at a Nissan garage and they said they should be pushed out with the force of the water coming through. Possibly blocked or kinked pipes stopping the water coming through and preventing the washers from coming out. Just wondering if anybody has any thoughts. The fact that both of them aren't working i was thinking more like a fuse or something rather than both pumps being gone but i cannot find a fuse just for the headlamp washers. The windscreen washers and headlights themselves are working fine.
Any thoughts or suggestions would be very welcome.
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Re: Headlight Washers Not Working

Postby DaveBerlin » Tue Apr 05, 2016 5:33 am

Nial, I am assuming that your QQ is the J10 Model. I found this older comment on the other QQ Forum perhaps it will help perhaps not but for the Guy with the problem it did work - Dave ;)
2014 1.2l DIG-T QQ Tekna QAB Pearl White (M)-delivered on 08.14./Premium Pack Chrome /Style Pack Chrome/Cross Over Pack Front & Rear in Pearlescent White/Auto-Fold Mirrors Chromed/Shark Fin Antenna/Bonnet Struts/Innen LED's/White Chrome Side Mouldings
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Re: Headlight Washers Not Working

Postby Arkangel » Wed Nov 30, 2016 8:35 pm

Hi Nial,
did you ever solve this problem with you QQ headlamp washer jets??
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Little boy invents something that could save your baby's life

Rubber band inventionYou guys, I am so proud of 11-year-old Andrew Pelham and I don't even know him, but I know this kid is smart and compassionate because of an invention he came up with in an effort to save babies' lives. His invention is called the E-Z Baby Saver. He's not trying to make any money off of it, all he's trying to do is save babies' lives.

He's created something that anyone can afford and make. In fact, it's so simple to make and use that he offers instructions on his website on how you can make your own.

Read more ¿Qué más?: Preschooler helps solve home invasion & you'll NEVER believe who planned everything

The invention came about because Pelham entered the Rubber Band Contest for Young Inventors, where it was required that contestants use rubber bands in their invention. Pelman was inspired to make something to help parents remember they had a child strapped in a car seat after learning that approximately 38 children die each year after being left in hot cars.

He says, "I'm hoping if I can save one life with my invention, I can say that my invention was successful." I love this kid, I really do. He's got some kind of a wonderful heart at such a young age.

The invention is basically a simple strand of rubber bands and duct tape that you attach to the back of the driver's seat and when you have a child strapped in the back, you attach the front part of the strand to the driver side's door. When the driver goes to get out of the car, they can't without first unhooking the E-Z Baby Saver and thus being reminded that they have a child in the back.

Check out Andrew Pelham and his awesome invention, the E-Z Baby Saver! Oh and by the way, his invention got second place in the contest and he won $500. Nice.

Image via Thinkstock

What do you think of this little boy and his invention?

Topics: parenting  safety  inspiration

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has been here before.

In late 1999, Amazon's stock price vaulted into the triple digits, fueled by exuberance over the promise of e-commerce. But by 2001, slammed by the double whammy of a recession and the popping of the Internet bubble, the stock had collapsed back into the single digits.

Now, Amazon shares are back in the stratosphere, propelled this time by improved profit margins and swelling revenues.

On Tuesday the company announced a third-quarter profit of $80 million, a threefold increase from the $19 million it earned in the third quarter of 2006. It reported sales of $3.26 billion, up 41 percent from $2.31 billion in the quarter last year.

In advance of the earnings report on Tuesday, Amazon's stock price gained 10 percent and topped $100 for the first time since December 1999, closing at $100.82. In after-hours trading the stock gave up most of those gains, trading around $90.

Scott Devitt, managing director of Stifel Nicolaus, a financial services firm, said investors might have been worried that Amazon would start to increase its spending next year.

''Expectations got ahead of themselves,'' Mr. Devitt said. ''You have a stock that has gone from $30 to $100 in a matter of 15 months, and at some point you crossed fair value.''

The company's strong results for the quarter can be partly explained by currency fluctuations and by the 2.5 million copies of ''Harry Potter and the Deathly Hallows'' that Amazon sold.

But investors and analysts are also encouraged by more enduring trends, like the migration of smaller retailers onto Amazon's network.

''Amazon's third-party business is on fire,'' said Scot Wingo, chief executive of ChannelAdvisor, which helps smaller retailers sell their wares on the site. ''People that dipped their toes in the water with Amazon a year or two ago have really expanded their selection.''

Unlike eBay, Amazon does not charge a listing fee for each item sold on the site. Instead it charges only a commission, taking an average 10 to 15 percent cut of each sale. That allows sellers to experiment with all of their merchandise on Amazon and pay only when an item is sold.

Amazon beat Wall Street's quarterly estimates of $3.14 billion in revenue and its own forecasts of $3 billion to $3.175 billion. Mark Mahaney, the director of Internet research at Citigroup, said investors had generally been pleased by a slowdown in Amazon's investment spending at the same time that margins and revenues soared.

''EBay's margins are flat on the year, Google's are flat to down on the year, and Yahoo's margins are flat to down,'' Mr. Mahaney said. ''You go to Amazon and revenue growth is more than 30 percent, along with a margin expansion of 25 percent. Compound those two and you get earnings growth this year that is just dramatic.''

Amazon's investment in its own operations is still considerable: it spent $181 million on what it calls technology and content development over the last three months. But the rate of that investment is slowing and the investments are starting to yield fruit.

In September, the company introduced the Amazon MP3 digital music store to sell tracks without the antipiracy technology known as digital rights management, or D.M.R. The music companies EMI and Universal are participating in Amazon's store, making the service a significant competitor to Apple's iTunes service.

In a conference call with analysts, Jeff Bezos, chief executive of, said the company was happy with early results from the store. ''We are getting terrific feedback from customers,'' he said. ''Everybody loves the D.R.M.-free format. Now the onus is on us to continue to convince music labels that this is a good way to sell their music.''

This fall, Amazon is also expected to introduce an electronic book device and an online store where users can download e-books.

Another nascent but promising area for Amazon is its Web services business, in which it rents out parts of its computing infrastructure to other Web businesses.

More than making up for those developing businesses is the overall increase in e-commerce sales. The research firm Hitwise reported double-digit increases in traffic to Internet retail sites during the summer of 2007.

Amazon got the largest chunk of that traffic, with 11.5 percent of all visitors, followed by Wal-Mart, with 5.4 percent.

Amazon also raised its forecast for 2007, saying it expected to take in $14.3 billion to $14.6 billion for the year.

Graph: ''Rescaling the Heights'' Amazon's shares are near highs last reached in 1999. (Source by Bloomberg Financial Markets)


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Brooks B15 Swallow Chrome

B15 Swallow Chrome

Availability: Check Store Availability
  • This item is not permitted to be shipped, however it can be purchased online and picked up at one of our stores at your convenience OR we can deliver to your home or office for a $50 fee to the following counties in Maryland: Anne Arundel, Baltimore, Carroll, Harford, Howard, and Baltimore City.
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Brooks B15 Swallow Chrome

Brooks' B15 Swallow has been described as the precursor to all modern racing saddles. When it was first patented in 1937 it became immediately popular among the professional racers of the era, and has remained imminently popular amongst cyclists who prefer a sporty, performance-oriented saddle. Like every Brooks saddle, the B15 Swallow shapes itself to its rider over time, ensuring the timeless, customized support that the company's reputation is built upon. And, of course, the B15 Swallow features Brooks' full-grain, premium tanned leather and trademark rivets for the ultimate in comfort, style, and durability.

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China's Elite Politics provides a new theoretical perspective on elite politics in China and uses this theoretical perspective to explain power transfer from Jiang Zemin to Hu Jintao and political dynamics between different factional groups since the Sixteenth Party Congress of November 2002. It explains the transition in structural terms, presents characteristics of China's political elites, and analyzes the balance of power among formal institutions as well as among factional groups. It also examines political interactions between Jiang Zemin and his cronies on the one side and Hu Jintao and his allies on the other over a number of issues: the epidemic of severe acute respiratory syndrome (SARS); ideological institutionalization; the politics over economic overheating; Jiang Zemin's complete retirement; and Hu Jintao's power consolidation in both ideological and personnel terms.

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From the man who energised legions of people with the 80/20 phenomenon comes a proven, practical programme for anyone who wants to achieve extraordinary results without extraordinary effort. With his first mega-bestseller, The 80/20 Principle, Richard Koch applied a century-old economic principle--80 per cent of results come from 20 per cent of our efforts--to show companies how to drive business performance. The 80/20 Individual showed how to release the potential of the 80/20 principle in the workplace. Living The 80/20 Way applies Koch's less is more and more with less ideas to your best 20 per cent in everyday life: how to get work you enjoy, how to build stronger relationships with family and friends, and how to unmask the mystery of money.

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Real Estate

Real Estate and Homes for Sale in Island Park, NY

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New Jersey Real Estate Lawyers

New Jersey Real Estate LawyersIn New Jersey, real estate law is a very broad area of law covering anything that has to do with the ownership of land and buildings.

Real estate law can apply to many numerous facets of a personal and professional life. It can apply to anything from the purchase of a vacant lot, to renting prime downtown office space.

New Jersey real estate law can be very intricate, because it's really a collection of a lot of different areas of law. These areas of law include property rights, contracts, land use, and many others.

Having a good basic knowledge of New Jersey real estate law can make a lot of transactions, such as buying a house or renting an apartment, much easier.

» Find New Jersey Real Estate Lawyers Now

Find Real Estate Lawyers and Law Firms in the Largest NJ Cities

Show New Jersey Cities

Common Real Estate Issues in New Jersey

Financing: Most people in New Jersey probably can't afford to buy a house or a piece of land up front. Consequently, most individuals and business use some form of financing to purchase real estate, typically a mortgage. A mortgage is a loan used to buy a piece of real estate, with the purchased real estate simultaneously securing the loan.

Disclosure: When a house or other structure is for sale, the seller has to tell prospective buyers about all physical defects of which the seller is aware, and which the buyer couldn't be reasonably expected to discover. However, if you are selling a house, you should probably just disclose every defect you're aware of. This will save you a great deal of potential liability. Even if your disclosure of a particular defect wasn't required (because the buyer could have easily discovered it), the buyer might still sue, and the cost of defending this lawsuit, even if you win, will be very high.

Warranty of Habitability: This applies in residential leases. Landlords are under a legal duty, whether it's specifically stated in the lease agreement or not, to ensure that the rented property is fit for human habitation. The requirements for habitability often include electricity, heating, running water, a working phone line, and protection from the elements. Of course, that list is not exhaustive.

Zoning Laws: Zoning laws dictate what kinds of buildings can be placed on lots in certain parts of a town or city. Their purpose is to ensure, for example, that sewage treatment plants don't get built next to homes, or that noisy, polluting factory doesn't get built next to a daycare center. To this end, they "zone" sections of the locality for numerous uses, such as residential, industrial, commercial, etc. If you are buying a piece of vacant land, it's essential that whatever use you intend to make of it is compatible with New Jersey's zoning laws.

Do I Need a New Jersey Real Estate Lawyer?

These matters can be pretty convoluted, and the above discussion only scratched the surface of the huge body of law that can affect real estate transactions. Therefore, if you think any business or personal plan you have might be affected by real estate law, you should consult with a New Jersey real estate lawyer as soon as possible.

Find the Right Lawyer Now

New Jersey is located in the northeastern quarter of the United States and has a population of well over 8 million people. It is the most densely populated of all the 50 states and is listed as the third wealthiest according to median household income.

New Jersey's legal system revolves around its three basic levels of courts- the Municipal Courts, Superior Courts, and the New Jersey Supreme Court. Unlike most states, New Jersey does not have an intermediate court district for appeals; instead, the Superior Court operates its own Appellate Division that processes appeals. There is also a Tax Court system with limited jurisdiction to settle tax disputes.

The judiciary of New Jersey is also unique in that it still has separate courts for law and equity. Also, the Superior Court system is further divided into divisions of Law and Chancery at the trial level. These features allow the state to process a variety of different claims in an efficient manner. New Jersey has also produced many major figures in U.S. jurisprudence, including U.S. Supreme Court Justices William Brennan, Jr., Antonin Scalia, and Samuel Alito.

Lawyers in New Jersey have experience in guiding clients through the state's court system. Most New Jersey lawyers begin by filing claims at the Superior Court level. Attorneys are available to provide legal advice and representation in New Jersey courts.

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Thursday, November 15, 2012

Reposting: Aman Futures and the 3rd Deadly Sin

I feel the need to repost this as hubby and I now are both beginning to invest and work on with our funds and hard-earned money. This is a must-read. Please do. Be wise and be safe.

Read more HERE.

The first rule of money, according to lay evangelist and entrepreneur, Bo Sanchez is, “If you earn it, don’t lose it.”

The caveat would seem useless now for thousands of people who plunked their lifetime savings in a get rich scheme run by Aman Futures Group Philippines, Inc. The Pasay City-based trading firm thrived in Pagadian City until the managers could no longer pay investors. The police are looking for Aman Futures top guy Manuel Amalilio and his cohorts who have all quickly disappeared. Meanwhile, Interior and Local Government Secretary Mar Roxas assured investors the government will do everything to collar Amalilio and recover their money.

I commiserate with the victims, but my readings about pyramiding scams tell me that recovery would translate to the Tagalog saying, “Pagputi nang uwak, pag-itim nang tagak.” (When the crow turns white and the heron turns black).

Get rich schemes, known as pyramiding, also known as Ponzi, are nothing new.
One of the biggest investment scandals that rocked the country was the operation run by Multinational Telecom Investors Corp. Multitel’s Rosario “Rose” Baladjay duped not just ordinary people but also businessmen, politicians, Cabinet members and celebrities who lost an aggregate of P25 billion to the scam.

Although the massive investment rip-off was discovered in 2002, Multitel ran an underground operation as early as 1988. Investments as low as P2,000 and as high as P10,000 earned a monthly interest of 1 percent or 12 percent per annum. Baladjay lent the money to businessmen at a rate of 2.5 percent a month or 30 percent per annum.

The lending investor hit the jackpot when it offered huge interest rates of 30 percent to 40 percent in 45 days. Double your money schemes offered 80 percent to 100 percent returns if the investment is locked in for 18 months. In 2002, the pyramid collapsed under the weight of clients’ demands for profits. Baladjay was convicted for the crime of syndicated estafa and was sentenced to 12 years imprisonment and maximum penalty of life imprisonment.

Some people think Pinoys fall easy prey to pyramiding schemes because they are presented like investment and stock markets plans. This is partly true.

In July 2009, I wrote about Bernard L. Madoff and how his Ponzi-style investment caper ripped off an estimated $65 billion from reputable banks and investment houses in the United States and Europe, including celebrities, retirees, and charitable institutions.

The key to getting their trust was Madoff’s credentials because he was a skilled stock broker and financial adviser who later went on to become chairman of the US stock exchange NASDAQ. The moneyed class viewed him as a modern day Midas, whose touch turned any investment portfolio worth bars of pure gold. Madoff ran the enterprise for years and delivered on the promise of 30 percent to 40 percent profits within 45 days, or 100 pecent returns within 90 days.

The term Ponzi is attributed to the con man of the ’40s, Charles Ponzi who was able to pay off dividends by using funds from early investors. But yet, Ponzi was not even original because in 1899 there was William F. Miller, a Brooklyn accountant who pocketed $1 million through the pyramid scheme.

With such important lessons from the past, we wonder why ordinary people, but especially professional investors and financial experts would still fall for the scheme. We often hear the admonition to be wary with offers that sound too good to be true, but some investors practically pleaded with Madoff to accept their money.

And so the question is, was Madoff so glib that he was able to dupe investors and bankers, or were his victims so stupid that they were deceived by a financial plan with con game elements written all over it?
Was Amalilio’s driver and janitor such a sweet-talker that people waited outside the office of Aman Futures at 3 a.m. just to give him their money?

Far from it. The experience of peoples who lost everything in get rich schemes is rooted in the third deadly sin, greed. People are drawn to the promise their money will earn much “while they sleep.” Common sense tells them otherwise, but they don’t care how the investment planner would raise 30 percent to 100 percent interest for as long as the profits kept coming. They know the plan is bound to fail, but they were fed with initial sums, and they thought it would last a lifetime.

While Madoff’s victims pondered their fate in the spring of 2009, CNBC ran a series entitled, “The American Greed.” I had the privilege of watching one feature, about Texas billionaire and offshore banker Allen Stanford, who used practically the same scam in duping clients in the US and in Antigua. Stanford endeared himself to the island government in the Caribbean region by buying real estate and involving himself in philanthropic activities.

He was eventually knighted by the island state, and later allowed to buy a small island.
Stanford had wanted to corner the port services of Antigua but was derailed by a top department official who smelled something fishy.

As the Madoff caper unravelled, Stanford’s clients in the US and Antigua also lined up to withdraw their investments but as this predictable story turned out, they were not even worth the paper they were printed on. The CNBC series was supposed to bring to light the “dark side of the American Dream.”

Madoff’s saga ended in June 2009 after a federal judge sentenced him to 150 years in prison. Nothing could alleviate the pain of victims, but Judge Denny Chinn saw to it that the scam be remembered for all time, to serve as lesson and warning. He pronounced the crime as extraordinarily evil.

In concluding the article, I wrote that Hollywood celebrities like Steven Spielberg, who also lost millions in the Madoff caper, should be able to put a more thoughtful spin on the American Dream, “the supposed inner soul or essence of every American to achieve a better, richer and happier life.”

Being rooted in the US Constitution that all men are created equal, the national philosophy is an inspiration for many poor Americans, especially blacks and white immigrants, who rose from rags to riches through hard work and perseverance.

Sadly, these virtues that launched the US to the top of the world are waning and instead, there is a growing unscrupulous desire for more wealth and power.

This is a malady not just in American society, but in many cultures around the world including our own. As shown by the Madoff caper and Philippine versions, Multitel and Aman Futures, the value of hard work and sacrifice has lost its meaning, while material riches have become man’s obsession.

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Bond Selling Seems Forced

 | Aug 22, 2013 | 2:48 PM EDT
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It's worth noodling on who is selling bonds right now, because it sure does seem forced. With the help of my colleague, Matt Horween, let me give you some thoughts on the natural sellers and why they might be selling.

Foreign governments own a huge percentage of $15 odd trillion in U.S. government debt -- perhaps as much as 45%-50% of all bonds. China alone owns $1.2 trillion in bills notes and bonds. Japan owns almost a trillion in bonds. Brazil has $200 billion. Wow, those are some real weak holders. China's got to be dumping like mad and that could be a huge source of repatriated dollars for a country that's starting to awaken, as the inching higher Baltic Freight index shows and the purchasing manager's index, proves, given that it went north of 51 when last month's reading was 47. How can Brazil maintain its holdings right now? The country is totally strapped. And tell me that Japan doesn't want some of that money-- although they sure don't want to do anything to move up the yen, which is what would happen if they repatriate too aggressively.

Plus, why not sell? These countries aren't watching Abercrombie & Fitch (ANF)plunge off of a horrendous quarter and terrible guidance. They aren't saying "Wow, Macy's (M) is bad and Target (TGT) is worse." They are looking at weekly unemployment claims and saying, "Gee, things are heating up there. I want out." Or they are saying, "I can't take this parlor game of tapering or no tapering -- let me just book my huge gain and move on."

Remember, they might sense a real bargain in the euro. Two years ago, we thought the euro was going away. Now, given the newfound strength of even the periphery countries, you have to think that owning bonds denominated in euros could be a better play than dollar denominated bonds. Suddenly there are big bond market alternatives that make sense to savers.

And who can blame them? We have had a holiday from government dysfunction but that holiday is about to end. And does anyone even think for a minute that anything's changed? The wrangling in Washington is terrible for the bonds and, of course, it has caused all of the big declines in the stock market for the last few years.

It's coming back.

The bond market decline may seem like panic-selling to many Americans -- and there is an element of that, for sure -- but these are seasoned holders who have made fortunes and they don't want to give them away. They may have missed the bottom but they are nowhere near the top. They don't care if rates go back to 2.7%. They just want to take the money back, go elsewhere or ring the register. In fact, there are so many more reasons to sell than to buy, that you can see how bonds can overshoot any economic activity -- as I think they have already.

The natural sellers are headed home. Who can blame them?

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A taste of Chengde's Eight Dishes

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Ingredients Jump to Instructions ↓

  1. 1 pound(s) skinless, boneless chicken thighs , timmed of fat 2 teaspoon(s) vegetable oil 1 teaspoon(s) lower-sodium fish sauce 3 tablespoon(s) lower-sodium fish sauce 1/4 cup(s) fresh lime juice 1 clove(s) garlic , crushed with press 1/2 teaspoon(s) light brown sugar 1/4 teaspoon(s) Asian chili sauce (Sriracha) 1 head(s) green cabbage , trimmed and cored 1 medium red onion 3 hearts romaine lettuce 1/2 cup(s) packed fresh cilantro leaves , plus additional for garnish 1/2 cup(s) packed fresh mint leaves , plus additional for garnish 3 tablespoon(s) roasted unsalted peanuts

Instructions Jump to Ingredients ↑

  1. Heat large ridged grill pan on medium until hot. In large bowl, toss chicken thighs with vegetable oil and 1 teaspoon fish sauce until evenly coated. Working in batches if necessary, cook chicken 7 to 9 minutes or until juices run clear when pierced with tip of knife, turning once. Transfer chicken to plate. Cool slightly; cut into 1/2-inch chunks. Meanwhile, in large bowl, whisk lime juice, garlic, sugar, chili sauce, and remaining fish sauce until combined. Thinly slice cabbage and add to bowl with dressing. Toss to coat evenly. Let stand while preparing remaining ingredients. Meanwhile, thinly slice onion and coarsely chop lettuce, cilantro, and mint; add to bowl with cabbage. Chop peanuts and add to bowl, along with chicken and any juices. Toss to combine. Garnish with additional cilantro and mint.

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Ingredients Jump to Instructions ↓

  1. Ingredients (serves 4)

  2. 1 tbs olive oil

  3. 1 brown onion, finely chopped

  4. 1 carrot, peeled, finely chopped

  5. 1 celery stick, finely chopped

  6. 4 pancetta slices, chopped

  7. 4 cups (1 litre) chicken stock

  8. 400g can Italian-style chopped tomatoes

  9. 1 zucchini trimmed, finely chopped

  10. 1/2 cup (about 60g) small penne

  11. 1/2 cup (80g) frozen peas

  12. 400g can borlotti beans, rinsed, drained

  13. Basil pesto

  14. 1 cup basil leaves

  15. 1/4 cup (45g) toasted pinenuts

  16. 1 garlic clove, finely chopped

  17. 1/4 cup (60ml) olive oil

  18. 25g parmesan, coarsely crumbled

Instructions Jump to Ingredients ↑

  1. Heat the oil in a large saucepan over medium heat. Add the onion, carrot, celery and pancetta and cook, stirring, for 5 minutes or until onion softens. Add the stock, tomato and zucchini and bring to a simmer. Reduce heat to low and cook, stirring occasionally, for 20 minutes or until soup thickens slightly. Add the pasta and cook for 10 minutes or until tender.

  2. Add the peas and borlotti beans and cook, stirring, for 5 minutes or until heated through.

  3. Meanwhile, place the basil, pinenuts and garlic on a chopping board and use a sharp knife to coarsely chop. Transfer to a bowl. Add the oil and parmesan and gently stir to combine. Season with salt and pepper.

  4. Ladle the soup evenly among serving bowls. Spoon over the basil pesto and serve immediately.

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Recipes Wiki

Florentine Catfish

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Description Edit

A Catfish recipe.

Ingredients Edit

Directions Edit

  1. In a large saucepan, bring the water and lemon slices to a slow simmer.
  2. Add the catfish fillets and poach gently for about 8 minutes.
  3. Remove fillets and drain well.
  4. Thaw and drain the spinach.
  5. Place it in a kitchen towel and squeeze the moisture from the spinach.
  6. Set aside.
  7. Melt butter in a heavy saucepan.
  8. Add green onions and garlic.
  9. Cook briefly to soften.
  10. Add flour and stir constantly while the roux cooks.
  11. Do not brown.
  12. Slowly add the milk, continuing to stir until the sauce is smooth and thickened.
  13. Add reserved spinach, seasonings, lemon juice, and ½ cup of the cheese, mixing well (the sauce may be prepared ahead to this point and refrigerated until needed, up to 2 days).
  14. When you are ready to serve, place the cooked catfish fillets in a lightly buttered 9- by 3-inch baking dish or individual au gratin dishes.
  15. Cover with sauce and top with remaining cheese.
  16. Bake at 350°F for 15 minutes or until bubbly.
  17. Finish under the broiler to lightly toast the cheese.
  18. Serve immediately.

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1000 Strings Play For Christmas

The Sounds Of A Thousand Strings was the name of the background music orchestra that recorded for Crown record between 1958-1964. After that the recordings were attributed to groups such as "The Fascinating Strings". This group of albums that this orchestra made were made to compete with the 101 Strings orchestra, the group even a couple knock off albums of the 101 Strings charting "Soul Of Spain" albums (which the Sounds Of A Thousand Strings orchestra albums were titled "The Heart Of Spain"). This is the album was most likely a knock off the very popular 101 Strings "Glory Of Christmas" album. There are more selections from this album on my Youtube channel, go check it out!

Silent Night
White Christmas

  1. Probably the best known today of the "Sounds of a Thousand Strings" LPS is MUSIC FOR BIG DAME HUNTERS, with its sexy cover photo of Irish McCalla, aka TV's "Sheena of the Jungle," in a more daring version of Sheena's leopard skin outfit.

  2. One of the reasons the Irish McCalla 1000 Strings LP is so rare is because the Bihari Bros got sued for lifting the tracks from another LP (not the first time they got caught doing that). (The other reason it's so rare is that people kept the cover and threw out the LP.)
    I wonder where they lifted the tracks for this one?

    The red vinyl looks neat, and the cover design is perfect considering the fact that the holiday season was a really big sales period for rack-jobber LPs.
    It sounds like this LP is in terrific shape (or is it just these two tracks?).

  3. I just discovered your blog thanks to Lee Hartsfeld's blog.

    This is one of my all time favorite Christmas albums. I have been on a quest to find out who originally recorded it. As you probably know, it was reissued a ton of times with different covers and artist names. I discovered it as a child. My dad bought it on the Parade label as Al Goodman and his Orchestra. It was issued on the Spin-o-rama, Custom,and Premier labels among others. The artist later became George Jenkins & the Christmas Strings, who ever he was. Several years ago I found mp3 files of these recording for sale on Amazon or one of the other download sites under the name of "Art Neville" Thanks for your blog.

  4. I am curious to know if there a rar or zip file for this album?

  5. I am sorry to say that there is not a youtube recording of "Hark! The Herald Angels Sing" video from this record :o(

  6. I just recently lucked into a copy of the "Big Dame Hunters" LP. Just gorgeous! And the record inside ain't too bad either... In the "who?" category, Crown credits "Marc Collins" as the conductor of the 1000 strings; but then, they managed to misspell Irish McCalla's name at the same time. To KL From NYC: Do you know whose album the Biharis swiped the tracks from? If yes, hope you will share this info.


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Blogs > Red Cards

Bob Weiser started playing soccer as a youth and has been following the sport ever since, dating back to days of watching the Bundesliga on PBS on Saturday afternoons.

Monday, July 19, 2010

All Union, all the time

The Union had an impressive week, getting its first shutout, or clean sheet, with a 1-0 win over Celtic FC of Scotland in the first night game at PPL Park and following that up with a 2-1 win over Toronto FC Saturday on Sebastien LeToux's late penalty kick.

Of course, it should be pointed out that that first shutout came with backup keeper Brad Knighton in goal, not starter Chris Seitz. I've been saying for weeks that Knighton deserves a shot to play over Seitz, whose inconsistencies and inexperience have cost the Union on a couple of occasions. I was disappointed to learn, then, that Seitz was back in goal Saturday against Toronto.

But the good news is they got 3 points against Toronto, a team that had an eight-game unbeaten streak heading into the game. Toronto, by the way, features Jon Conway, the Sun Valley High School graduate who is the only Delaware County high school product playing in MLS. See Chris Vito's story here:

Despite temperatures in the upper 90s again Saturday, LeToux was all over the field. His work ethic is simply amazing.

Up next is a friendly against Manchester United Wednesday night at Lincoln Financial Field. Man U will be holding a clinic Tuesday night from 7-9 at Chester Park for local youths. The Union's next MLS match is against the New England Revolution July 31 at PPL Park.

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August Entry-Level Referee Clinic

Posted - July 1, 2013

Redding Youth Soccer League will be hosting an entry-level referee clinic in August.  The successful student will have taken 18 hours of instruction on the Laws of the Game and pass a US Soccer Federation exam of 100 questions.  Upon successful completion the new referee will be able to work youth games anywhere in the USA. Certification will be valid  through December 2014, at which time an annual certification fee of $40 will be due.

The cost for this clinic is $25.  If the certification exam is passed, there is an additional $40 fee payable to the State referee association.  There are twenty spots currently available and a Wait List will be formed, if necessary.  Minimum age is twelve (12) years old on August 31, 2013.   If you are interested and available, please REPLY to me and I will send specific sign-up details.

  • Tues., August 13, 2013:  5:30 pm to 8:30 pm
  • Wed., August 14, 2013:  5:30 pm to 8:30 pm
  • Thurs., August 15, 2013:  5:30 pm to 8:30 pm
  • Tues., August 20, 2013:  5:30 pm to 8:30 pm
  • Wed., August 21, 2013:  5:30 pm to 8:30 pm
  • Thurs., August 22, 2013:  5:30 pm to 8:30 pm

Redding Soccer Park
Conference Room
9800 Old Oregon Trail
Redding, CA 96003

I look forward to hearing from you and welcoming you into the world of youth soccer refereeing.

Don Chin

Upcoming Events

No upcoming events found.

RYSL has two main registration periods. January for the Spring Season and May for the Fall Season of play. We notify all members via direct mail, the website, email, newspaper and other media outlets of the upcoming registration events.

We get asked some of the same questions on a regular basis. We've tried to cover the most frequent ones on the FAQ page. Everything from season length to what your child needs to play. Perhaps you can find your question and answer here? If not, feel free to drop us a line!

To provide a program dedicated to develop and sustain a quality, consistent, safe and positive environment to assist youth soccer players in attaining their highest possible level of personal skills, tactics, sportsmanship and overall soccer proficiency through proper teaching and competition. The clubs philosophy is that development of players and how we play the game is a greater priority than winning.

PO Box 991523; Redding, CA 96099-1523  |   Sorry, no phone number available.  |   registrar@reddingsoccer.com  |   social/email.png   social/facebook.png   social/twitter.png


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Light & Heavy Tackle
Sunshine Coast
Marlin Fishing Charters

Owner Colin Hombsch has been skippering charter vessels for 15 years.

The vessel Reel Action is a 15.5 metre cat powered by twin 1050 Volvo IPS engines and was custom built for marlin fishing!

Charters depart from Mooloolaba on the Sunshine Coast

Reel Action Charters offers you full day, light or heavy tackle billfishing options: