text:
- Research article
- Open Access
Comparison of hybridization-based and sequencing-based gene expression technologies on biological replicates
© Liu et al; licensee BioMed Central Ltd. 2007
- Received: 23 November 2006
- Accepted: 07 June 2007
- Published: 07 June 2007
High-throughput systems for gene expression profiling have been developed and have matured rapidly through the past decade. Broadly, these can be divided into two categories: hybridization-based and sequencing-based approaches. With data from different technologies being accumulated, concerns and challenges are raised about the level of agreement across technologies. As part of an ongoing large-scale cross-platform data comparison framework, we report here a comparison based on identical samples between one-dye DNA microarray platforms and MPSS (Massively Parallel Signature Sequencing).
The DNA microarray platforms generally provided highly correlated data, while moderate correlations between microarrays and MPSS were obtained. Disagreements between the two types of technologies can be attributed to limitations inherent to both technologies. The variation found between pooled biological replicates underlines the importance of exercising caution in identification of differential expression, especially for the purposes of biomarker discovery.
Based on different principles, hybridization-based and sequencing-based technologies should be considered complementary to each other, rather than competitive alternatives for measuring gene expression, and currently, both are important tools for transcriptome profiling.
- Microarray Platform
- Mouse Retina
- UniGene Cluster
- Biological Theme
- Mouse Cortex
During the last decade, a considerable number of high-throughput technologies for transcriptome profiling have been developed. These include hybridization-based technologies, such as DNA microarrays [1–3], and sequencing-based approaches like SAGE (Serial Analysis of Gene Expression) [4] and MPSS (Massively Parallel Signature Sequencing) [5]. The power of DNA microarrays lies in the simultaneous hybridization of mRNA extract from biological samples to a pre-selected mRNA library, which can contain up to tens of thousands of various mRNA transcripts. The expression levels of each transcript are obtained by reading out intensities of hybridization signals. Sequencing-based methods are based on a substantially different strategy as compared to microarray technologies. SAGE and MPSS do not require any pre-compilation of an mRNA library of sequences, but instead, they use type IIS restriction endonucleases, i.e. tagging enzymes, to collect short tags (typically 10–22 bases) from each mRNA molecule, provided a relevant recognition site exists for an anchoring enzyme. Then, either by sequencing long concatamers of tags using conventional sequencer (SAGE), or by performing iterative parallel sequencing using a proprietary technique (MPSS), the identity of a sufficiently large amount of tags can be determined in an efficient manner. The abundance of each mRNA transcript is assumed to be proportional to the count of occurrence of its representative tag.
Favorable features of hybridization-based approaches include a significantly lower workload and relatively low cost. However, the probe collection on a chip, which necessarily relies on the coverage and the accuracy of both genomic sequences and clone libraries, presents a hard constraint on its detection power. In contrast, the "tag-and-count"-based methodologies require more advanced instruments that are more cost- and labor-intensive, but their capability of exhaustive transcript sampling allows the potential identification of novel mRNAs.
The present co-existence of various DNA microarray platforms and sequencing-based technologies offers biomedical research increased options for transcriptome profiling. It is, however, important to understand how to compare data generated by these different technologies. Recent studies indicate that performance of various microarray platforms, as measured by data consistency, have been shown to be comparable [6, 7]. Several attempts have also been made to compare heterogeneous types of technologies, for instance, between microarray and SAGE [8–15], and between microarray(s) and MPSS [16–20]. The results of these studies demonstrate moderate concordance between technologies.
In the present study, which is part of an ongoing cross-platform comparison framework [6], we compare gene expression data from MPSS with data from five different commercially available one-dye microarray platforms. The present study extends our previous study in three ways: 1) the inclusion of the Illumina BeadArray® and MPSS data, 2) the inclusion of gene expression data of a second pool of mouse retina (MRP2) for all microarray platforms, and 3) the investigation of variation across biological replicates, as measured over two different pools of mouse retina samples (MRP1 and MRP2). It has been a consensus that the use of biological replicates is an important element to ensure the reliability of microarray results [21]. To our knowledge, this is the first study investigating the differences between microarray and MPSS data on biological replicates. MPSS libraries are usually constructed without technical replication and data variation across samples are generally estimated by applying a statistical model simulating the random sampling process during tag selection for sequencing [22]. It remains unclear whether significant variation across samples examined by MPSS is comparable with those detected by microarrays. As our study included technical replicates for the microarray platforms, this provided a unique opportunity to investigate the sampling model by comparison to microarray data where technical replicates permitted a more robust statistical testing. The data sets of microarray data for the first mouse retina pool (MRP1) and mouse cortex (MC) were analyzed in our previous study, but some results on these are also included here for the sake of performing comparisons between the two mouse retina pools, as we believe the investigation of biological replicates is an important and novel aspect of this study.
In summary, our results showed that there were moderate, yet significant, correlations between microarray data and MPSS data, while the data from microarray platforms, including the recently included Illumina arrays, generally were well correlated. The majority of discrepant measurements between the technologies based on hybridization versus sequencing were genes with low-abundance transcripts. Tag-to-gene mapping ambiguity and the absence of tagging enzyme recognition site could also explain some of the discrepancy between MPSS and microarrays. Using two-way ANOVA and SAM, we examined the microarray data for the magnitude of data variation introduced by biological replicates and technical replicates, and showed that biological variations are smaller than platform variations. The genes we found most susceptible to variations between biological replicates were likely to be associated with metabolic pathways, biosynthesis, and binding related pathways. Due to their vulnerability to sample variation, any changes observed in these pathways and related genes should be interpreted more cautiously in biomarker discovery applications. Furthermore, as demonstrated in our study, the relative affordability of array replicates makes them useful to corroborate MPSS experiments where technical replicates are seldom feasible due to higher costs. For comprehensive transcriptome profiling, we suggest that complementary use of hybridization-based and sequencing-based technologies is likely to provide a better solution than pursuing a single type of technology alone.
Characterization of MPSS data
MPSS libraries for MRP1 and MRP2 were generated using 'Signature' MPSS protocol. Two alternative sequencing reactions conducted independently, i.e. two-stepper and four-stepper sequencing, provided two read-outs of tag sequences for each sample, and referred to as MPSS 17-bp and MPSS 20-bp, respectively.
In the 17-bp signature sequencing, a total number of 34,341 signatures were detected for MRP1, and 29,509 signatures for MRP2. The total number of signatures was obtained in the 20-bp signature sequencing was 34,424 and 30,967, for MRP1 and MRP2, respectively.
The assignments of signature to gene were performed as described in the Methods section. Only the reliable MPSS signatures were kept in the downstream analysis. In the 17-bp signature libraries, 6,001 and 5,340 unique UniGene identifiers for MRP1 and MRP2, respectively, were identified. This corresponds to the sum of transcript copies as 615,944 and 615,771 tpm (transcript per million). For the 20-bp tag collections, the numbers of unique UniGene identifiers were slightly smaller: 5,793 and 5,125 for MRP1 and MRP2, respectively, which corresponds to the sum of transcript copies as 655,919 and 652,588 tpm.
Characterization of microarray data
All platforms showed generally good consistency among technical replicates for all samples, both in terms of CVs and correlation coefficients (see Additional file 1). These two metrics did not reveal noticeable differences between the two mouse retina mRNA pools within each platform.
In order to compare data from microarray platforms, we calculated both Pearson and Spearman correlation coefficients for absolute expression levels and relative expression changes. To transform expression measurements from diverse platforms onto a common scale, percentile transformation was applied to absolute expressions for each array data set, and then the median of percentile transformed intensities across replicated measurements per gene was used (see Methods for details). Filtering was applied but was not observed to increase the correlations between intensities, while the correlations between log2ratios were considerably improved with filtering.
The inter-platform data agreement by measuring correlation coefficients is shown in Additional file 1. The correlations between Illumina and the other platforms were generally lower than between other pairs of microarray platforms. This could possibly be due to the lack of technical replicates for Illumina.
Comparison between microarrays and MPSS
Statistics of overlapped genes between microarray and MPSS
Statistics on overlapping genes between microarray platforms and MPSS libraries.
MPSS (17-bp)
MPSS (20-bp)
MPSS (17-bp)
MPSS (20-bp)
MPSS (17-bp)
MPSS (20-bp)
Correspondence between microarray and MPSS
Pearson correlation coefficients across data sets
MPSS (17 bp)
MPSS (20 bp)
MPSS (17 bp)
MPSS (20 bp)
Furthermore, we found that 305 genes (286 in MRP1 and 272 in MRP2) were present across five microarray platforms, but were not detected by MPSS, neither from the 17-bp library nor the 20-bp library. For both libraries, about 64% of these genes were filtered out in the tag-to-gene mapping procedure. A total of 97 genes were not detected by MPSS at all (any sample or library), among which 11 were labeled "GATC"-negative. Using percentile-transformed data, we found that 67.8% of the remaining genes (60 of 86) were detected as having low expressions (percentile less than or equal to 50). This indicates that low-end sensitivity is the major reason for data discrepancy.
Taking the whole MPSS libraries as gold standard, we defined false negative detections by each microarray as the number of genes that were detected by MPSS, as well as printed on the chip, but were not detected by the microarray. We found that the proportion of false negatives in the five microarray platforms ranged from 12.99% (Affymetrix, MRP1) to 2.18% (ABI, MRP1).
Evaluation of data variations in biological replicates
Using SAM to characterize variation in detection of expression changes across biological replicates
Two-way ANOVA to identify individual genes with significant variation
Two-way ANOVA analysis was performed to identify the contribution of platforms and samples on data consistency. We examined filtered log2ratios (MRP1::MC and MRP2::MC) after normalization matched by both RS (RefSeq) and RSEXON (RefSeq ID and exon) across all four platforms where technical replicates were available. Only those genes that did not have missing values across the 10 measurements were used in this analysis. In RS- and RSEXON-based matching, 563 and 305 genes, respectively, satisfied the criterion. The threshold of statistical significance was chosen as p < 0.001.
Summary of two-way ANOVA results
total # of genes (i.e. RSs or RSEXONs)
# of genes that showed neither sample- nor platform- related variation
# of genes that showed sample-related variation
# of genes that showed platform-related variation
# of genes that showed both sample- and platform- related variation
Comparison of data variation between microarray and MPSS
From the four microarray platforms with technical replicates, we used one-way ANOVA to identify genes with differential expression in MRP1 versus MRP2. These results were then compared with the Z-test statistic commonly used for detecting differential expression in MPSS data. The microarray data used for this analysis was restricted to genes that had valid measurements for all ten technical replicates. We also excluded measurements from probes with ambiguous UniGene mapping, which was used to match microarray data with MPSS data. For the four microarray platforms, Affymetrix, Amersham, Mergen, and ABI, we obtained 4,783, 5,987, 3,610, and 8,460 UniGene identifiers with corresponding p-values from one-way ANOVA, respectively.
Comparison of differentially expressed gene identification between microarrays and MPSS
MPSS (17-bp)
MPSS (20-bp)
MPSS (17-bp)
MPSS (20-bp)
MPSS (17-bp)
MPSS (20-bp)
MPSS (17-bp)
MPSS (20-bp)
Number of genes included in the comparison
Number of genes considered as differentially expressed in both microarray and MPSS
Number of genes considered as having NO differential expression in both microarray and MPSS
Number of genes that are considered as differentially expressed in MPSS but not in the microarray
Number of genes that are considered as differentially expressed the microarray but not in MPSS
GSEA to summarize differences between biological replicates using biological themes
For each microarray platform, we applied GSEA to generate hypotheses regarding which biological processes or pathways might be responsible for the differences between the two replicate sample pools. Only the genes (RSs) which had valid values across all 10 chips were used, that is, 2,490 for Affymetrix, 4,899 for Amersham, 3,132 for Mergen, and 7,556 for ABI. In GSEA, the GO hierarchy level was set to 4, and the permutation times as 1000. Several biological themes resulted from GSEA with enrichment score, but none were statistically significant. Although different microarray platforms revealed different degrees of variation between MRP1 and MRP2, the GSEA analyses on the affected gene sets were similar among platforms. When considering enriched themes found in all four platforms, there were 15 GO terms from the "biological process" category enriched in MRP1 compared to MRP2 and 12 GO terms from the "molecular function" category. There were no terms found to be enriched in MRP2 common to all platforms [see Additional file 3].
With microarray technology being rapidly developed and advanced in the past decade, it has become an important tool for studying gene expression patterns. In parallel, the technologies based on the "tag-and-count" principle, such as SAGE and MPSS, are also being used for exploring the full transcriptome. Although some efforts in designing and adopting standards of microarray experiments (e.g. ERCC [24]) and data deposition (e.g. MIAME [25]) are paving the way towards data meta-analyses and integration, it remains a critical challenge to systematically compare cross-sample, cross-platform, and cross-technology data. To this end, we have established a framework which accommodates various platforms and various technologies, using quality-controlled biological samples [6]. Several recently published studies [7, 23, 26] have concluded that, for DNA microarray technology, the reproducibility of technical replicates both within a given platform and across platforms are generally good, especially when the experimental design, protocols, and data analyses are standardized [6]. In this study, we have examined data correspondence and discrepancy between MPSS and one-dye microarray platforms, representing sequencing-based and hybridization-based technologies, respectively. We examined whether and how the behavior of these distinct technologies would vary when challenged with biological replicates.
The main observations made in this study were that across the microarray platforms, both intra- and inter-consistency of data was generally high, but the agreement between MPSS and any microarray platform was moderate, as also reported in previous studies [17, 18]. The differences in signal detection between MPSS and microarray platforms were observed both in terms of lower correlations and in terms of genes that were consistently found as expressed in MPSS but not in microarrays and vice versa. These findings were also identified in a similar study [18], where this could be the reflection of the known limitations of the MPSS technology [27]. Although mapping problems may have contributed to the observed discrepancies, it is more likely that inherent differences between hybridization-based and sequencing-based technologies caused the systematic differences in gene expression detection between MPSS and microarrays. For a given microarray design, the set of genes that can be detected is pre-determined, while for MPSS and similar technologies, the main limitation is that presence of a recognition site is a requirement for detection. However, when comparing the genes represented on the various microarrays to the genes detected as expressed with MPSS, we found approximately 80% of the genes on the ABI and Illumina arrays had not been found in the MPSS libraries and about half of the genes for the Affymetrix, Amersham and Mergen arrays. This suggests that all the microarray designs were comprehensive with respect to genome coverage, and that the fixed probe sets may not have been a main limitation in this study. Furthermore, in all microarray platforms there were a number of genes that were detected as present but not found in the MPSS libraries. Among the 97 such genes (UniGene IDs) common to all five microarray platforms, there were 11 that were found to be lacking the DpnII recognition site ("GATC") and could not be expected to give any signal in MPSS. For the remaining 86 genes, the expression measurements on the microarrays varied and included genes classified as consistently highly expressed, as well as genes classified as having consistent low expression. An investigation of the probe sequences of the most highly expressed genes in the microarray platforms did not reveal noticeable differences in the number of possible sequence matches between those found in MPSS and those not found in MPSS. It is nevertheless possible that some of the false-positives in microarrays relative to MPSS could be caused by cross-hybridization due to suboptimal probe design. However, without a gold-standard, it is not possible to ascertain that the microarrays are overestimating the expression for these genes. For the genes identified as having lower expression in microarrays but not found in MPSS, it is possible that this may have been caused by running the MPSS experiments with insufficient sampling depths resulting in a less representative sampling of tags. There were also a number of genes that can be regarded as "false-negatives" on the microarrays relative to MPSS, in the sense that they were represented on the microarrays and detected by MPSS but not detected as expressed by the microarrays. For these genes there was less consistency across the platforms and only one gene detected by MPSS was not identified as expressed by any of the microarrays where it had been represented. Again, suboptimal probe design due to incomplete sequence knowledge can be a factor. Other possible reasons include MPSS sequencing errors [19], high complexity in transcriptional activities [28], heterogeneity in polyadenylation cleavage sites [29] and various sequence-introduced biases [30, 31]. We have however not been able to estimate the size of such contributions in this study. It has also been reported that the existence of SNPs [32] can influence the interpretation of digital-based experimental data such as MPSS or SAGE, but this is not expected to have been a major contributing factor in this study. The fact that the mapping to UniGene IDs had been based on two different versions of the UniGene database is a possible confounding factor. However, this cannot explain all the discrepancies as several genes were manually checked against the latest UniGene build and found to have consistent mappings.
Data filtering is commonly applied both to microarray and MPSS data. Both in the present study and an earlier study, we have shown that data filtering considerably improves data consistency between microarray platforms, and in particular on relative expression (log2ratios). The low intensity signals generally corresponding to low-abundance transcripts are typically filtered as it is expected that the signal-to-noise ratio becomes too small. The comparisons between MPSS and microarrays indicate that the MPSS technology also has problems in detecting low-abundance transcripts. It appears that neither technology can reliably detect transcripts expressed at very low frequencies in an environment where the expression levels of all transcripts could span several orders of magnitude. This is a major problem as many transcripts have low-abundance, and there is currently a consensus that many of the corresponding genes are associated with critical regulatory roles in the cells [33]. For microarrays to improve the low-end sensitivity, the optimization of probe design, as well as the advances of scanning techniques, may be key issues. In sequencing-based systems, increase of library size and sequencing accuracy may increase the confidence in low-abundance transcripts identification.
Recent studies have demonstrated that factors such as strain [34, 35], gender [36, 37], as well as diet [38, 39] and circadian variation, [40] can influence gene expressions in various organisms and tissues. In our study we created two retina samples by randomized pooling of samples from a large number of individuals aiming to cancel out the effects of such factors. We examined the biological variability at several levels: (1) the influence of biological replicates on the measures of intra- and inter-platform data consistency; (2) the overall capability of differentially expressed gene identification by each biological replicate; and (3) the genes or gene sets that were most susceptible to biological variability between MRP1 and MRP2. In general, the two pools showed very similar gene expression patterns as expected, but for some genes there was a difference between the two mouse retina pools that was consistent across the platforms and technologies. Moreover, MPSS also differed considerably from the microarray platforms in terms of identification of genes differentially expressed in MRP1 versus MRP2. As is commonly done due to high instrumental complexity and cost, MPSS data for each sample was collected without technical replicates, and differentially expressed genes were identified by a statistical approach. The Z-test, found by Man et al. [22] to perform well in terms of specificity, power, and robustness for determining statistical significance in SAGE, detected far more differentially expressed genes than the statistical tests applied to the microarray data with technical replicates. Also in terms of fold-change, MPSS detected far more genes as having two-fold or larger change than the microarray platforms. In this respect, Illumina data, which also did not have technical replicates, behaved similarly as the other microarray platforms, hence the lack of technical replicates alone cannot explain all of these differences. In light of the construction of the two retina pools and the good agreement between the microarray platforms, this may be a sign of caution for those who intend to use MPSS for the purpose of biomarker discovery without incorporating technical replicates. Until technological advances make technical replicates in MPSS feasible and MPSS data are further studied and confirmed using independent and complementary technologies, MPSS may not be the optimal choice for identification of novel fingerprints based on differential expression.
The GSEA results across the microarray platforms with technical replicates indicated some common biological themes for the differences between MRP1 and MRP2. Although the results were not statistically significant, they were very consistent across the platforms and indicated metabolism and transcriptional regulation as general biological themes describing the differences. This was also confirmed by GOstat [41] analysis using a list of genes consistently identified as differentially expressed when also including MPSS data (data not shown). Based on the construction of the two retina pools, these results indicate that caution should be exercised when interpreting results of differential expression.
An earlier study [42], which compared microarray data, EST-based expression experimental data and SAGE using published data sets, reached the conclusion that the agreement between the methods was highly variable from gene to gene, and the authors advocated the need for gene-by-gene validation of important global gene expression measurements using non-global methods. The present results support a similar conclusion, and we emphasize that sequencing-based methods in general as well as hybridization-based methods have inherent technological limitations. Microarrays are limited by pre-selected gene sets and possible cross-hybridization problems, as was indicated in this study. Apart from the obvious limitations of restriction site presence, MPSS is an open-ended system but has problems related to the mapping of tags limiting the set of genes for which it is possible to obtain reliable measurements. Altogether, this suggests that exploitation of the complementarity of these technologies is a better approach for global transcriptome analysis.
Overall, the agreement between MPSS and microarrays was significant, but lower than between different microarray platforms. Measurements of genes with low expression more often disagreed than highly expressed genes, as expected, but also for genes with high expression there were systematic differences in detection. We found that differences in gene expression measurements between MPSS and microarrays are not only due to increased sensitivity of MPSS to low abundance transcripts and the ability of MPSS to measure new transcripts. Further studies comparing sequencing-based and hybridization-based technologies, including both biological replicates using different types of tissue samples as well as technical replicates are warranted in order to delineate in more detail the shortcomings of these technologies. Future methodological development will be necessary to maximize the information derived from the two complementary types of technology.
Biological samples
RNA samples were isolated from three sources: two pools of C57/B6 adult mouse retina (MRP1 and MRP2, n = 700) and Swiss-Webster post-natal day one (P1) mouse cortex (MC) (n = 19). Retinas were dissected, collected and stored in Trizol (one pair of retinas per eppendorf tube) at -80°C prior to pooling. During the RNA extraction process, two pools of adult mouse retina (MRP1, MRP2) were created (700 retinas per pool), aliquoted. All samples were stored at -80°C until being used in experiments. The animal experiments were approved by the Institutional Animal Care Facility at Harvard University.
Microarray platforms, data processing and consistency assessment
Whole-genome mouse gene expression arrays (one-dye oligonucleotide microarrays) were investigated in this study, including: Affymetrix GeneChip®, Amersham (now GE Healthcare) CodeLink®, Mergen ExpressChip®, Applied Biosystems (ABI) microarrays, and Illumina BeadArray®. Microarray experiments are composed of sample preparation, hybridization, scanning and image quantitation, which are a series of integrative procedures being conducted at a laboratory, generally according to the manufacturer's recommended protocols. To obtain sufficient statistical confidence in the data analysis, for each biological replicate (MRP1 and MRP2), five technical replicates on each platform were obtained, with an exception on Illumina. We wanted to include Illumina in this study to examine the magnitude of data variation in MPSS experiments and microarray experiments when no technical replicates are performed. For details of the experimental protocols and laboratories, we refer to Kuo et al., [6] except for Illumina, which can be found in the Additional file 4.
The raw data sets of 63 chips after image scanning and quantification in each platform were collected. For Illumina data, we set the filtering threshold as "Detections" = 0.9. Filtering for the other microarray platforms are described in Kuo et al. [6]. We also performed percentile transformation of intensities, quantiles normalization and log2ratio calculation, as described.
Data repeatability and reproducibility [7] are two important aspects of microarray data consistency assessment. In this study, the former will refer to the degree of data variations among technical replicates of a platform, and the latter will refer to data agreement across different microarray platforms when using the same biological samples. Two popularly used metics, coefficient of variations (CV) among replicated measurements per gene and correlation coefficient (Pearson and Spearman correlations) between any pair of replicated experiments, were adopted to assess repeatability and reproducibility. For intra-platform data consistency, the mean and standard deviation of CVs or correlation coefficients were used as summations of each platform's performance. For inter-platform data agreement, either the mean (for normalized log2ratios) or the median after percentile transformation (for intensities) of repeated measurements on each platform were used in calculating correlation coefficients.
MPSS experiment and data processing
Total RNA of MRP1 and MRP2, which were identical to those used in microarray experiments, was sent to Lynx Therapeutics, Inc. (now Illumina, Hayward, CA) for 'Signature'-based MPSS experiments. Following an RNA quality test on a Agilent 2100 BioAnalyzer (Agilent Technologies, Palo Alto, CA), cDNA libraries were generated according to the Megaclone protocol [5, 43]. Signatures adjacent to poly (A) proximal DpnII restriction sites ("GATC") were cloned into a Megaclone vector. The resulting library was amplified and yielded about 1.6 million loaded microbeads, which were loaded onto a flow cell. Thereafter, an iterative series of enzymatic reactions decoded the signatures as 17-bp or 20-bp sequences (including DpnII recognition sites "GATC") [44].
The abundance of each signature was converted to transcripts per million (tpm), and the MPSS signatures were mapped to UniGene clusters by Lynx Therapeutics, based on the mouse genome sequence (Release #3, Feb 2003) [45] and the mouse UniGene sequences (UniGene Build #122) [46]. Briefly, the mapping procedure included: extraction of 'virtual' signatures from genomic sequences, classification of 'virtual' signatures from genomic sequences, and matching of MPSS expressed signatures to genomic signatures [44]. For the comparison with microarray data, we included only the reliable signatures which were located closer to polyadenylation signal or poly(A) tail on a mRNA sequences with known orientation information [47]. If a UniGene cluster was found to be corresponding to multiple signatures in a given library, all tag counts were pooled to obtain the abundance of the UniGene cluster. If a tag was found to map to multiple UniGene clusters, the corresponding tag count was discarded.
Gene mapping among microarray platforms and between microarray and MPSS
Two approaches to match probes across different microarray chips, annotation-based and sequence-based probe matching were used [6]. Briefly, by the annotation-based approach, we obtained UniGene (UG) and LocusLink (LL) based matching, whereas probe matches at the RefSeq (RS) and RefSeq-exon (RSEXON) levels by utilizing actual sequence information belong to the latter.
MPSS signatures were mapped to UniGene clusters, using an in silico constructed "virtual tags" library, as described above. Thus, the gene expression data measured by microarrays and by MPSS were paired up for comparisons via UniGene clusters.
Biological variations and technical variations
Two separate total mRNA extraction processes were conducted on mouse retina under the same experimental settings and protocols, which generated mouse retina pool 1 (MRP1) and pool 2 (MRP2). Note that Illumina data were not included in this part of analyses due to lack of technical replicates.
Two-way ANOVA to identify individual genes of significant variation
To investigate the effects of biological variation and platform variation, two-way ANOVA (Analysis of Variance) was performed. As concluded in our previous study [6], the sequence-based cross-platform probe matching is more reliable than the annotation-based probe matching. Therefore, RS- and RSEXON-based mapping were used for this evaluation. For a given set of transcripts (RSs or RSEXONs) that were reliably detected in all chips of all platforms, the significances of sample-dependent bias, platform-dependent bias and interaction between sample and platform were determined for each transcript. Thus, we were able to observe the gene-specific effects of sample and platform biases.
SAM to characterize biological replicates' behavior in detecting expression changes
A group of differentially expressed genes represents the desired result of most microarray users. SAM (Significance Analysis of Microarray), proposed by Tusher et al., [48] is a method that can determine the significance of gene expression changes by permuting replicated measurements followed by an estimation of the false discovery rate (FDR). SAM assesses both the sensitivity and specificity of a microarray platform.
In our study, for each platform, the five pairs of chips on which MRP1 and MC were hybridized respectively were considered as one experiment group, while similarly another experiment group consisted of MRP2 and MC data. Prior to SAM analysis, the normalized log2ratios underwent two sequential filtering steps: (1) filtering according to spot quality flags; and (2) filtering out the probes which had less than three valid measurements out of the five replicates. For each microarray platform and each experiment group, the number of "called" genes (as differentially expressed) and FDR were recorded for every threshold "delta", which steps from 0.1 to 4 at an interval of 0.1.
GSEA to summarize differences between biological replicates using biological themes
GSEA (Gene Set Enrichment Analysis) [49, 50] combines functional annotations and a statistical mechanism to determine a few sets of genes, each with a biological theme, which are over-/under-represented between two data sets representing two different classes. Instead of looking at per individual gene, GSEA is focused on gene sets pre-defined according to Gene Ontology (GO) category, pathway, or localization, etc, whose results have more robust and explicit biological interpretation. We used GSEA, a web-based application provided by Babelomics [51], to identify which biological process(es) and molecular function(s) were the most susceptible to the random differences between MRP1 and MRP2. The GSEA results of each platform were also compared.
One-way ANOVA analysis of microarray replicates and Z-test of MPSS runs
One-way ANOVA was conducted for each microarray platform (10 experiments: five for MRP1, five for MRP2). One-way ANOVA uses those genes that have valid measurements (filtered intensities after normalization) across all experiments as input, and assigns a p-value to each gene indicating whether this specific gene displayed significantly varied expression levels between two biological replicates.
MPSS analyses were also performed on the two pools of retina sample independently. Due to its high cost, it is not feasible to conduct repeated MPSS experiments for each biological sample. The common practice for those who use MPSS to identify differentially expressed genes has been to apply statistical tests that are able to handle non-replicate data based on certain sampling assumptions. The Z-statistics test [22] was applied to identify genes that were significantly differentially expressed between MRP1 and MRP2.
Accession numbers
The microarray data and MPSS data for the manuscript has been submitted to the GEO Omnibus. The series record number is GSE6313.
We would like to thank Applied Biosystems, GE Healthcare, Mergen, and Lynx (now Illumina, Inc.) for providing and running the experiments as part of this large scale study. WPK was supported by the National Institutes of Health (NIH) EY014466 grant, the Bioinformatics Division of the Harvard Center for Neurodegeneration and Repair and Harvard School of Dental Medicine Dean Scholars award. CLC was supported by the Howard Hughes Medical Institute. FL and EH were supported by the functional genomics program (FUGE) in the Research council of Norway.
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Swinton Lions 18-25 Rochdale Hornets
Sunday 3rd June 2018, 15:00pm
Championship - Round: 2
Rhodri Lloyd 2, Chris Hankinson 2,
Chris Hankinson,
Danny Yates, Gary Middlehurst, Rob Massam, Deon Lewis Cross,
Tyler Whittaker 4,
Drop Goals
Tyler Whittaker,
Line Ups
Swinton Lions
Rochdale Hornets
Match Report
Rochdale Hornets climbed off the bottom of the table with a 25-18 victory over Swinton Lions at Heywood Road on Sunday.
It was the perfect revenge for their Summer Bash defeat at the hands of their local rivals just one week earlier, as a number of senior players returned to help them to victory.
Gary Middlehurst, Toby Adamson, Luke Adamson and Ben Moores all returned from injury, while Jack Fox made his debut at centre after joining the club on a trial basis.
A late flurry from the hosts had Hornets worried in the dying stages of the game, but a Tyler Whittaker field goal gave the visitors some breathing space.
The halfback also kicked four goals, and was brilliant in the victory as he led his side around the field and finished sets well with the boot.
Jonah Cunningham gave his best performance in a Rochdale shirt, playing a pivotal role coming off the bench at hooker to compliment the returning Moores.
Although there were some standout individual efforts, it was a true team performance as Alan Kilshaw's side showed a huge amount of guts to bounce back from last week's disappointing defeat.
It took 13 minutes for the away side to break the deadlock, as a Middlehurst offload went to ground and was scooped up by Whittaker, who broke through the middle and sent Danny Yates over in support.
From the kick-off, Lewis Hatton spilled the first possession of the set and his side were punished, as the ball was put through hands with Rhodri Lloyd eventually crossing.
It was then Swinton's turn to knock on at the restart, as Middlehurst made the most of the opportunity that error presented by spinning over from close range.
Half an hour into the game Rob Massam gave Hornets a 14 point cushion with an individual effort, as he ran from dummy half and found the line with a low drive despite the attentions of three Lions defenders.
With eight minutes to go in the first period, Josh Barlow was sent to the sin bin for running into a melee following a late shot from Pat Moran.
The half came to an end without any further scores, as the visitors took an 18-4 lead into the break.
Six minutes into the second half, Hornets scored the try of the game as a Mike Butt knock on gave them a set in a dangerous position, which ended in Deon Cross rounding off a brilliant flowing move by racing through a gaping hole on the left edge.
The hosts' response came from Chris Hankinson, as he found some space out wide after three sets in a row on the Rochdale line.
On 70 minutes Whittaker missed a penalty goal, and the fact it didn't go out of the field of play proved costly as Swinton went up the opposite end of the field to score.
Straight from a scrum George Tyson chased down a kick, but was hauled down just short of the try line by Richard Lepori, only for Lloyd to collect his second as he scored a walk-in effort on the next play with the defence still retreating.
That set up a tense finish, but the home side were a man down for the final four minutes when Barlow was red carded for dissent.
Whittaker's field goal kept Swinton at arm's length, as some desperate defence was final broken when Hankinson touched down his own kick which had collided with the post.
Hornets had a seven point lead with just one minute left on the clock, and they managed to see out the contest and pick up a vital two points.
LIONS: Fell, Butt, Hankinson, Lloyd, Tyson, Billsborough, J. Hansen, Bracek, H. Hansen, Austin, Thornley, Paisley, Hamlin. Subs: Barlow, Shelford, Morrall, Gallagher.
Tries: Lloyd (15, 73), Hankinson (55, 79) Goals: Hankinson 1/3, J. Hansen 0/1
HORNETS: Kay, Lepori, Fox, Cross, Massam, Whittaker, Yates, Taira, Moores, Middlehurst, T. Adamson, Mitchell, Allen. Subs: Cunningham, Hatton, Moran, L. Adamson.
Tries: Yates (13), Middlehurst (19), Massam (28), Cross (46) Goals: Whittaker 4/5 Field goal: Whittaker (78)

subset: commoncrawl


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Wall sconce: metal socket
Special version of our wall sconce with sockets in brass, copper or nickel. One of the earliest pieces we made that we have only improved over time with new colors and options.
Attaches to standard electrical boxes with a universal cross bar and cap. Hand made out of steel and coated in a durable powder-coat finish.
Made to order in 1-2 weeks.*
Light bulbs sold separately here.
UL listed for damp or dry locations.
*Orders of higher quantities may not fall in within the listed lead times. Please contact us regarding lead times on orders with higher quantities.
5" diameter backplate
6" projection from wall to end of socket
8" from top to bottom
110v or 220v wiring
75 watt incandescent, 9 watt CFL or LED
LED compatible
Bar Keeper's Friend or metal polish can be used to shine brass and copper parts.
Light bulb sold separately here.


text:
Bamco Home
5TH Wheel Hitches
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Police in Craigavon are probing an attempted burglary in Parkmore.
A pane of glass beside the door was smashed and a garden shed was entered though nothing was taken.
The incident happened sometime between 2pm on Friday, 27 March and 1pm on Saturday, 28 March. Police are urging those with information to contact them in Portadown on 101.


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Basic Structure of Heart
The heart is small organ about the size of one's fist, located in the middle and slightly to the left of the mediastinum in the chest. The wider top part is the base and the blunt tip projecting forward and to the left if the apex. The heart is enclosed by the pericardium which is thick, fibrous and non-elastic. It consists of two layers: the inner layer-visceral layer and the outer parietal layer. The two pericardial surfaces are separated by a pericardial space that contains approximately 10-20 ml of thin, clear pericardial fluid which acts as a lubricant and reduces friction produced by the pumping action of the heart.
Epicardium-an epithelial single-celled layer surrounding the heart, the innermost layer of the pericardium. Myocardium is the actual heart muscle layer composed of striated muscle fibers. The myocardium is responsible for the energy created to eject a volume of blood. Therefore, it is thicker in the ventricles than in the atria. Endocardium is the inner layer and consists of endothelial tissue lining the entire interior of cardiac chambers and covers the heart valves.


text: On Line Student Competition Local student wins first place in Online Art Competition. Dayton High's Andrea Pusatero won for her watercolor painting, "Brown Stone". 2nd and 3rd places went to two Henderson, NV, Coronado High students, Herle Librando for her oil, "Shaping the Playground" and Taylor Murphy for an oil, "Trust in Nature". The paintings will be on display for the run of the new Imagination Show, now through June 24, 2016. "Brown Stone" by Andrea Pusatero "Shaping the Playground" by Herle Librando "Trust in Nature" by Taylor Murphy


text:
Entry Detail
Metropol Security
08-Sep-2013 19:1:57
Contact Number:
+ 44 (0)20 78370111
Metropol provides a variety of security and business support services to both large and small businesses. We cover a range of security roles with the aim of providing a complete professional security package so your customers and employees can feel safe and secure at all times. An integral part of our service is helping to enhance your public image by providing presentable, courteous and customer orientated security staff, giving you added piece of mind. Our support and control network operates 24 hours a day to provide a dedicated programme of early recognition and intervention to your business. Our service is not a one-size- fits-all security product; it's a customised, targeted solution that takes into account many characteristics of your business, including location, image and clientele, backed up by proper implementation, management and monitoring. We pride ourselves on offering the following as standard: A proactive, accountable approach A positive, problem-solving attitude A discreet presence that blends with your existing operation and works with it, not against it Courteous, presentable and highly trained staff Rigorous training for all our staff, and yours if you require it Operations managers on call 24 hours a day, 7 days a week Reassurance that your business will be protected around the clock by people you can trust
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A wonder goal from debutant Omer Riza, a fine team effort and a dubious offside finish left Maidenhead United with nothing to show from their efforts as they were beaten 3-0 at home by Boreham Wood.
Riza, who boasts a career that started at Arsenal as a teenager, before starring for Trabsonspor in the Turkish Super League is Ian Allinson's latest addition to the side and the 32-year-old made an immediate impact.
This was Wood's best away result of the season against a battling Magpies side. Their lack of dominance came about thanks to a quite superb performance from Luke Garrard, who was in a class of his own, coupled with the resolute Elvijs Putnins. The keeper has now tasted defeat just three times in 13 appearances this season.
A nervy start produced few goalscoring chances, with Maidenhead's Leon Solomon coming close with a rising drive that went just over the bar.
Charlie O'Loughlin battled against the equally well-framed centre forward Alex Wall throughout, but picked up his tenth booking early on which will lead to a two-match suspension.
David Bryant hit a weak shot at Aaron Lennox, before opening the scoring after 24 minutes.
A chipped ball over the defence was helped on into the path of Bryant by Riza and with the entire Magpies back line looking to the referee, he volleyed home wide of Lennox with no flag offered by the assistant referee.
The collective bemusement to that goal turned to collective admiration to Wood's second three minutes later.
An interchange of passes between Mario Noto and David Hutton out on the right led to a perfectly threaded low ball across goal from the Wood captain, finding Riza just inside the penalty area wide on the left. With seemingly nothing on, he took a touch before executing a stunning curling finish into the top right-hand corner of the net with almost no backlift whatsoever.
The home side were suitably contained by Garrard and his men up until the interval when Wall almost skewed a clearance into his own goal, which would have ended the game as a contest.
Wood continued to dominate after the interval, with Lennox forced to save from Bryant and then Hutton after he was played in by Riza.
The new arrival continued to shine when Noto played him through between two defenders. He was strong enough to make the edge of the area where John Scarborough made a fine saving tackle. However the loose ball fell for Hutton, whose fierce shot was well held by Lennox.
There followed a period of sustained pressure from Maidenhead as Johnson Hippolyte rang the changes, using up all his options from the bench.
Leigh Henry headed just wide at the near post from a corner, before great defensive work from Daniel Brathwaite and Garrard prevented further work for Putnins.
Jermaine Hinds found himself in acres of space at the far post from a corner yet took an age and fired wide before Wall tried his luck from 35 yards but Putnins remained as reliable as ever.
O'Loughlin then committed a clumsy challenge which referee Mr Knight took an age to deliberate over, due to the mob protest from a number of animated Magpies. It was their boisterousness which probably saved O'Loughlin at the outset.
Another wild miss from Hinds signalled the end of the home side's chances and Wood mopped up to end the game in some style.
Osei Sankofa, winning the ball by the corner flag in his own half, steered it into the path of Sam Lechmere. His long ball down the wing was superbly brought under control by Riza, who awaited the overlapping run of Sankofa. He fed Bryant who switched play back wide for Lechmere to deliver a floating cross that Bryant headed down into the path of Sankofa. The right-back looked every part a forward bringing the ball down before sweeping it beyond Lennox into the corner of the net. It was his first ever goal for Wood.
Riza made way for Mauro Vilhete with two minutes remaining and the Barnet loan player saw two shots saved as Wood looked to run riot in the closing stages.
Allinson said: "Our aim now is to finish top of the chasing pack. Eighth is not out of the question and would be a great springboard to go into next season's campaign.
"We have played well against the top sides yet they always have quality on their benches whereas we are trying to bring on quality from within. Now we face a string of games against sides of our level where we can dominate."
On his latest addition, the manager added: " You never know what to expect when you introduce a 32 year old.
"Omer has shown that he has legs, stamina, a fine footballing brain and a real hunger for the game. He combined well today with David Bryant and his goal was a different class.
"That said, I thought Luke Garrard was immense today and it was probably his best game for us."
Boreham Wood: Putnins, Sankofa, Brathwaite, Garrard, O'Loughlin, Beckles, Hutton (Lechmere 71), Cox, Bryant, Noto, Riza (Vilhete 88). Not used: Jones, McCann and Lewis.


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It always psyches me to come across fantastic, amazing and courageous Deafblind people – my heart fills up with inspiration. I am sure thousands of people out there feel the same way.
Be inspired when you read about the awesomely wicked adventures of a Deaf mountain climber and adventurer, Heidi Zimmer. What's even more inspiring is that this lady has Usher Syndrome (Retinitis Pigmentosa) and doing all these challenging activities with a very limited peripheral vision is just mind-blowing.
Word has it is that Ms. Zimmer has this dream to scale all of the seven major peaks on the world. You go, girl!
I hope that you will share this story with many Usher Syndrome and Deafblind youth and adults, and tell them – if this lady can do it, so can they scale their dreams and make it out on top!
*hands wave and feet stomping for Heidi*
View a Youtube vlog by filmmaker Jose Saldana:
Go here to see Heidi's personal webpage:
Deafblind Mountain Climber, Heidi Zimmer
Tactile love and let's go climb a mountain with our canes


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Can't Determine What You're Looking For? Optimize Your SearchesOptimizing your site for search engine listings is only a method of determining a method to enhance your website's traffic and visibility. This operates by researching the favorite search terms that apply to your blog, called keywords, and taking advantage of those terms inside your site's content as well as other text.
Advertising is not the main objective when you use SEO methods to gain rankings for the site. You may get a boost in traffic, nevertheless it doesn't improve your ranking.
If you are looking to draw attention to your site or blog, post comments about the relevant blogs of others. This catches their attention and definately will a minimum of lead to some traffic. From the best case scenario, you can also end up with a backlink and a recommendation through the other blogger.
Search engines like google can only search exactly what is actually inside your website, rather than exactly what is inside an image description. Try to stay away from image description. Instead you ought to put a text box beneath the actual image so the words might be looked for in a search engine.
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Start through the top. You likely will not be able to tackle all of your website for optimization in one week, much less per day. Begin with your top ranked pages, and move down after that. The most popular pages could be the ones to bring the most hits from optimization anyway, but optimizing them all will take your website ranking upwards by and large.
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Tags: 2007, CA, California, east bay, Graffiti, Grone, Groner, oakland, OSCR, Osker, West Coast
This entry was posted
on Friday, February 3rd, 2017 at 12:00 pm and is filed under Photos, Rollers.
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The other day my husband told me, "I want biscuits, can you make biscuits?" I laughed, of course I can! And you can to by following this simple recipe. Skip the canned biscuits from the store and go fresh next time. You can also freeze these for future meals.
P.S. These smell soooo good fresh out of the oven.
What you need:
- 1 3/4 c. flour
- 1/4 t. salt
- 2 t. baking powder
- 1 1/2 t. sugar
- 1/4 t. baking soda
- 3 T. butter
- 3/4 c. buttermilk
Picture of the ingredients. For reference I use turbinado sugar and buttermilk powder in this recipe.
Mix together the dry ingredients.
Add in the butter and the buttermilk. Mix well.
Knead a few times until you have a nice dough.
Flatten the dough out with your fingers. It should be about 1/2 inch thick.
Use a biscuit cutter to make the biscuit shapes. I don't have a biscuit cutter so I just use a glass. You should be able to get 9 biscuits out of this recipe.
Place them on a baking sheet at bake at 350F for about 10 minutes until brown on top.
Serve with butter or jam. Yum!
Or if you want something with a little different flavor, you can try this other easy homemade biscuit recipe made with cream!


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ADDvantages Homeschooling Program
for Children with ADHD/ADD
Problems Solved for Homeschooling Parents
More and more parents are choosing homeschooling for children with ADHD/ADD and learning challenges. Many have taken this route out of frustration and desperation over watching their child become unhappier with each passing school year. Homeschooling can be a great option for some of these parents. Most likely you'll need some professional help along the way with children coping with these difficulties.
Regardless of the strategies and techniques you use, your child will continue to struggle and master academic achievement when the underlying cognitive processing skills are weak. Once these skills are remediated your child will benefit from your instruction and homeschooling. The good news is your child can flourish at home!
Problems are Now Solved with ADDvantages!
Problem: Your child gets frustrated and "shuts down."
Solved: Your child is learning easier and faster.
Problem: Your child needs repetition to learn.
Solved: Your child remembers taught lessons.
Problem: Your child refuses to cooperate
Solved: Your child is motivated and eager to learn.
Problem: Your child is easily distracted.
Solved: Your child is better able to focus on the task at hand.
ADDvantages for Homeschoolers
With ADDvatanges Learning Center's Homeschooling program, we provide your child with an opportunity to learn how to learn. As a homeschooler you have the time and flexibility to make a significant impact on your child's ability to learn and ADDvantages can help!.
There is hope with ADDvantages' programs.
Learn about our Assessment and how this first step can help you, your child or loved one experience success.


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Jiangsu Flying Dragon Food Machine Co,.Ltd. is a professional enterprise for developing, manufacturing and marketing candy machines and food packing machines, with solid technical foundation and large production capacity.
The products are sold all over the country. the products are sold all over the country and exported to the countries and regions in middle east. southeast Asia, South Africa, India, Russia etc. enjoying great reputation in all customers. "cultural inheritance, exploitation, innovation, profession... [Details]


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Picture of Porchetta From Porchetta Restaurant in New York
Savory Sight: Porchetta
Even if you don't eat meat, it'd be hard to disagree with YumSugar member ericairis and her assertion about porchetta below.Fabulous pork from Porchetta NYC. To die for!
Do you have an image of something delicious that you recently made or enjoyed? If so, be sure to upload it in the Savory Sights group in our YumSugar Community!


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Why is Art Important? (My Reblog from the DLC)
I wrote this post for the Discovery Learning Center last week and wanted to share it again here.
Why is Art Important?
The question is not an easy task for some. Art easily takes a back seat to reading, math, science, and even sports in education these days. And it certainly isn't on the latest test.
But ask yourself…
Can a lesson in composition resound with a future web designer, or a lesson on perspective later help with geometry? Or perhaps learning color theory will help a future physicist understand how light works? How about the visual and contemplative of any art piece that could inspire a poem, the next great work of literature, or any myriad of invention.
Art education is something that is as important in the classroom as it is in the real world. Art helps a person grow and to appreciate; to see the world from art's own creative perspective. It helps one become a more rounded and cultured human being . Art isn't just for the "artsy" either; any person can learn to appreciate art. Creating a piece of art can make a person find something in themselves they have not seen before, regardless if they have "talent" or not.
Art education creates thinkers, innovators, problem solvers, communicators, humanitarians. It nurtures the development of self-esteem, self-discipline, cooperation, and self-motivation. Art helps a person to understand the human experience and helps to shape our identity.
Even the great innovators of the past and of today know and understand the importance of art as a part of education.
"In my own philanthropy and business endeavors, I have seen the critical role that the arts play in stimulating creativity and in developing vital communities….the arts have a crucial impact on our economy and are an important catalyst for learning, discovery, and achievement in our country."
–Paul G. Allen, Co-Founder, Microsoft
"It is in Apple's DNA that technology alone is not enough—it's technology married with liberal arts, married with the humanities, that yields us the results that make our heart sing."
–Steve Jobs, in introducing the iPad 2 in 2011
"Logic will get you from A to B. Imagination will take you everywhere."
–Albert Einstein
The Discovery Learning Center runs many art and creative based classes from fine art to tie dye. Join us for the next Art and Great Artists Friday August 24th at 10 a.m. Or check the calendar for the latest workshop.


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Seems I managed to finally settle for a Theme, Color schema and Typography – was hard enough to choose and funny that I finally choose a standard one.
Anyway, still need to figure out stuff about widgets, how to hide the Password protected pages without wrecking the menues.
But one step after the other.


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First Aired: July 12, 2015
The ways in which people's lives change for the better after cancer has everything to do with the body in which those wretched cells reside. So, there's no doubt that Jerri is a force to be reckoned with – a diagnosis of breast cancer in 2010 was followed by four surgeries and radiation treatments. She experienced the fears, frustrations and fatigue that come with cancer.
With the support of Cornucopia, her children and co-workers, she not only survived breast cancer, but used her illness to dig deep and cultivate her loving, brave self. Her smile is one of humble victory, a reminder of our ability to get beat up over and over and over and somehow emerge intact, victorious.
Hear Jerri's story of cultivating possibilities based on her inner sense of how she can make her way forward with gratitude, strength, peace, and compassion.
Return to That Cancer Show page


text: Riverbend Moving and Storage always offers free quotes for all your needs. We ask the important questions to give you a quick and competitively priced quote within 24 hours. You can request a quote using our easy to fill out quote form or over the phone. You can also set up an appointment to have one of our expert movers assess your items.


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MONEY | September 12th 2008
"As belts tighten, tempers flare", writes Joanna Moorhead in "Couples + Money". Simon Cox, a business writer for The Economist, examines the economics of coupledom...
From INTELLIGENT LIFE magazine, Autumn 2008
When companies face financial hardship, they are often tempted to merge, splitting their costs and combining their customers. Couples do the opposite. The weight of unpaid bills and overdue loans can break a relationship: the unspoken vow that haunts many marriages is "till debt us do part".
But a demerger makes little
economic sense. Ever since Gary Becker of the
It will surprise no one to learn
that men devote more hours to paid employment, women to housework. But contrary
to popular belief, these two contributions balance out, on average, in most
rich countries. In America, according to a report by Michael Burda of the Humboldt University of Berlin and others, men
do 313 minutes a day of market work and 163 minutes of housework (liberally
defined as anything you could pay someone else to do for you), whereas women do
201 and 271 minutes, respectively. The great exception is
The economic gains from coupledom are substantial. For middle-aged women, the cost
of living in a married couple is 30% less than the cost of living alone,
according to calculations by Arthur Lewbel of
But how do couples divide these
spoils? Economists once touchingly assumed that spouses wanted the same things
and acted as one to achieve them. Households were "glued together" in
Amartya Sen's phrase. Now
economists think of marriage as a bargain in a second sense. Husbands and wives
haggle over cash, chores, care and consideration. By one recent estimate, the
split in
Bad times can force a painful
renegotiation. Of unemployed men consigned to household chores, Friedrich
Engels wrote: "One may well imagine the righteous indignation of the
workers at being virtually turned into eunuchs." That indignation has
lingered in the 160 years since he wrote about it. Anne Solaz
These bargains may not be fair. But economists typically assume they are efficient. Between them, spouses create the biggest pie possible, even if they do not split it equitably. No other bargain could make one of the spouses better off, without making the other worse off. The traditional family, for example, strikes a bargain in which the wife sacrifices her career prospects to raise the children both spouses want, freeing the man to make more money. If the husband is sufficiently generous with his improved earnings, the bargain should make them both better off.
But this model of marriage is
becoming obsolete, according to Betsey Stevenson and Justin Wolfers,
an unmarried couple at the
Households also now spend less
time, and more money, feeding the family. But even in
The marriage factory faces a second threat: couples now find it harder to make their bargains stick, because vows can now be broken cheaply and without stigma. If a woman agrees to sacrifice her career to make a home, her prospects outside marriage diminish, leaving her with less conjugal clout. The husband might therefore be tempted to renege on his commitment to share his earnings equitably. But if his wife anticipates this outcome, she won't make the bargain in the first place. The rise of easier, "no-fault" divorces, Betsey Stevenson shows, has made American women less willing to have children or to pay their husband's way through college. And no one takes home economics anymore.
Coupledom has changed, these two economists argue, from a mode of production to a mode of consumption. Instead of complementing each other in their work, spouses complement each other in their pleasures. The husband enjoys cinema-going more because he goes with his wife. In the traditional economic conception of the family, one spouse provides the things money can buy, the other provides the time good housekeeping demands. Now a spouse is someone to spend time and money with. That may be some consolation when economic misfortunes leave a couple with more time and less money than they'd bargained for.
Picture credit: korbatz/flickr
(Simon Cox is a business correspondent for The Economist.)


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Four Visions for the Future of Housing
7 February — 17 May 2015
What might the future hold for housing in the UK? What are the challenges faced now and how can they be overcome? I put these questions to Mæ Architects, Dallas Pierce Quintero, 5th Studio and Sarah Wigglesworth Architects who came up with a range of intriguing visions which provided the basis for this well received exhibition. Informed by new and emerging approaches to housing and the way we live, the exhibition pointed towards a future for housing in the UK that is full of possibilities. Read more


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When Madonna's on tour, the local media are always given some footage of each concert to use in news reports and the like. Usually it's short clips from the first few songs of the show (on her 2012 MDNA tour it was a clip of "Girl Gone Wild" every night!).
Back in 1987 however, the media were generally not given this on the "Who's That Girl" tour - instead the press were allowed to come along with their own TV cameras to get the footage themselves!
We got to see the first couple of minutes of "Open Your Heart", "Lucky Star" and "True Blue" from many gigs on the tour, filmed each time by local TV channels. Usually this was via one fixed camera placed quite a distance from the stage.
Here's a rare treat - some of that original press footage, which has just recently been uploaded from the original tapes by their owner, in pristine quality. It's from Sunday 26th July 1987 - Madonna's concert at the Texas Stadium in Irving (a suburb of the city of Dallas).
(KellyWurx, July 1987)


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Oh my goodness - is there anything cuter than little boys and the teddy bears they love? Check out this great new read!This book is so much fun, has adorable illustrations, and has such a good lesson nugget inside! Thomas tries all of the strategies he can think of to keep a bear out of his pillow and blanket fort. Along the way, Thomas decides that the bear, his little brother, is more fun in the fort than out. Such a cute read.
Thursday, October 6, 2016
How To Share With A Bear - Eric Pinder- Mom About Library #5
3 comments:
Oh my goodness this sounds ADORABLE! I want to buy this for my little guy RIGHT NOW. I agree with you, little boys cuddling is basically THE best! Thanks so much for sharing this, perfect stocking stuffer for me to get!ReplyDelete
Such a cute book! This would be a great recommendation for all the parents I get coming in looking for books in preparation for a new sibling.ReplyDelete
Thank you so much for commenting! If you are a blogger as well, please be sure to leave a link to your blog- I'd love to visit!
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UK Visas and Immigration (UKVI) will introduce an easy visa process for Chinese business partners of UK companies. The scheme "Select Business Scheme UK" greatly reduces the paper work. Limits the documents to an invitation letter, a letter of employment, and a duly filled application form to apply for a UK visa.
The scheme will be accessible to China-British Business Council (CBBC), the British Chamber of Commerce in China, UK Trade and Investment (UKTI), and the Confederation of Business Industry (CBI).
The members of CBBC will be provided a welcome pack and a point of contact from UK Visas and Immigration department for any support, or queries. Plus, timely updates will be sent to all the participating members.
This shows the growing ties between the UKVI and Trade Bodies, said Dominique Hardy, UKVI's Regional Manager to China.
Source: Expat Briefing


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12/365 - Permeated within Senses
If you love an image (whether it's yours or someone else's) and want to make sure others get a chance to see it, you can "feature" it by choosing the "Spotlight" button.
When an image is Spotlighted, it receives enhanced visibility in premium spots throughout the site. Spotlighted images are rotated through these higher-visibility positions to ensure the best opportunity for the images to be seen by JPG users.
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Photo license: © All rights reserved
It was a good day to let life waft over me.......the scent of oriental lillies
Muliple exposure in camera
In the The Nature of Grounded photo essay.
6 responses
Pascal Fellonneau said (8 Jan 2010):
nice colours
Kathleen Mercado (Deleted) said (8 Jan 2010):
Beautiful warmth!:)
Alejandro González gave props (8 Jan 2010):
exelente !!!!!!!!!!!
Tricia Louvar said (8 Jan 2010):
Darn, I wanted to see another pair of your wicked cool shoes. Maybe tomorrow...;-)
I like the whispers of layers, if only flowers could use our language. BUT I imagine this is what a flower sees when it looks at you--beautiful.
Megan Green gave props (8 Jan 2010):
So very excellent.
John Linton gave props (12 Jan 2010):
Trippin'. The light, fantastic.
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Kornik is a family business with great understanding, love for wood and passion for carpentry.
We are makers of quality and bespoke furniture ideal for lofts and industrial spaces. Our furniture have authentic aged, factory and warehouse feel that would be beautiful and stylish addition to a home or a commercial space.
We can proudly say that all our products are designed, hand crafted and manufactured in Poland.
Offering high quality service and great attention to detail we strive to produce industrial furniture that excels commercial standard.


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Gambit (2013)
Director: Michael Hoffman
Cast: Colin Firth, Cameron Diaz, Alan Rickman, Stanley Tucci
Plot: An art curator decides to seek revenge on his abusive boss by conning him into buying a fake Monet, but his plan requires the help of an eccentric and unpredictable Texas rodeo queen.
Genre: Comedy / Crime
Awards: -
Runtime: 89min
Rating: PG13 for some suggestive content, partial nudity and a rude gesture.
IN RETROSPECT (Guest reviewed by Jason Lin)
It is not a given that a film will awe with highly acclaimed filmmakers' titles affixed to its credit roll. Here's a good example in Gambit, written by the Coen brothers (No Country for Old Men, 2007) and directed by Michael Hoffman (The Last Station, 2009). Adding on to the honorary list are actors Colin Firth, Alan Rickman and Stanley Tucci who render lukewarm performances toward the contemporary take of the 1966 original.
With a play of the pen, Joel and Ethan Coen have switched the setting of the film to Britain (and Texas) where the trickery ploy to cheat and get back against a wealthy art collector who can't determine art authenticity named Lionel Shahbandar (Alan Rickman) begins with a Harry Deane (Colin Firth) as well as an animated opening sequence in Pink Panther fashion.
For certain peculiar reasons, Deane allures a P J Puznowski from Texas to be part of his ploy together with Major Wingate (Tom Courtenay) who is a master art duplicator. The essence of the film should largely rely upon the onscreen interaction between these three characters, but to one's disappointment they fail to sizzle.
Less for a hilarious scene set within the middle act in a lavish Savoy Hotel that involves Firth roaming about the hotel bottomless. It is observed that critical screenwriting is the credit due for driving the segment to its full effect. To be honest, the screenplay by the Coens is technically sound and capable. It is a delight to witness how they induce plot points that connect to one another and how certain seemingly random details introduced will eventually make satisfying sense. These are hints of very capable screenwriters who do not introduce plot elements just for the sake of convenience.
However, the story and cast performances do little to inspire any further beyond the occasional laughing moments. Hoffman keeps an upbeat tempo to quickly flip along the story amidst overly sugarcoated dialogues. Some, if not most, of the conversational exchange feel excessively self-indulgent and redundant under opinion.
Throwing in a Texan accent sprouting Cameron Diaz as the individual with an exotic last name further complicates matters. This is where Tucci's brief role as a competing art authenticator Martin Zaidenweber feels under-developed, which is a pity as Tucci's German accent does wonders.
Summing up the film in further dismay is the questionable introduction of the stereotypical Japanese slapstick farce that begs viewers to ponder upon the integrity of the film material, which feels more in place in an exploitative slapstick comedy by lesser filmmakers.
Well, it is after all a rarity that the Coens write a film but not direct it. The filmmakers have certainly fared much better in their other works.
SCORE: 2 stars
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