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Expression level of many genes shows abundant natural variation in human populations. The variations in gene expression are believed to contribute to phenotypic differences. Emerging evidence has shown that microRNAs (miRNAs) are one of the key regulators of gene expression. However, past studies have focused on the miRNA target genes and used loss- or gain-of-function approach that may not reflect natural association between miRNA and mRNAs.
Methodology/Principal Findings
To examine miRNA regulatory effect on global gene expression under endogenous condition, we performed pair-wise correlation coefficient analysis on expression levels of 366 miRNAs and 14,174 messenger RNAs (mRNAs) in 90 immortalized lymphoblastoid cell lines, and observed significant correlations between the two species of RNA transcripts. We identified a total of 7,207 significantly correlated miRNA-mRNA pairs (false discovery rate q<0.01). Of those, 4,085 pairs showed positive correlations while 3,122 pairs showed negative correlations. Gene ontology analyses on the miRNA-correlated genes revealed significant enrichments in several biological processes related to cell cycle, cell communication and signal transduction. Individually, each of three miRNAs (miR-331, -98 and -33b) demonstrated significant correlation with the genes in cell cycle-related biological processes, which is consistent with important role of miRNAs in cell cycle regulation.
This study demonstrates feasibility of using naturally expressed transcript profiles to identify endogenous correlation between miRNA and miRNA. By applying this genome-wide approach, we have identified thousands of miRNA-correlated genes and revealed potential role of miRNAs in several important cellular functions. The study results along with accompanying data sets will provide a wealth of high-throughput data to further evaluate the miRNA-regulated genes and eventually in phenotypic variations of human populations.
Citation: Wang L, Oberg AL, Asmann YW, Sicotte H, McDonnell SK, Riska SM, et al. (2009) Genome-Wide Transcriptional Profiling Reveals MicroRNA-Correlated Genes and Biological Processes in Human Lymphoblastoid Cell Lines. PLoS ONE 4(6): e5878. https://doi.org/10.1371/journal.pone.0005878
Editor: Baohong Zhang, East Carolina University, United States of America
Received: February 12, 2009; Accepted: May 8, 2009; Published: June 11, 2009
Copyright: © 2009 Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported in part by a career development award from Mayo Clinic SPORE in prostate cancer, National Institutes of Health (CA91956 to LW) and in part by a grant from National Institutes of Health (CA126786 to LW). However, the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Expression level of many mRNA genes shows abundant natural variation in human populations. The quantitative variations in mRNA expression are thought to contribute to phenotypic differences between individuals. Several molecular mechanisms have been identified that control gene expression. In addition to known transcription factors that bind to specific regulatory DNA sequences [1], [2] and extensively studied genetic polymorphisms that determine transcription level via cis- or trans-effects [3]–[8], newly discovered miRNAs have been proven to be a major player in posttranscriptional regulation of gene expression [1], [2], [9], [10]. The miRNAs were first identified to play a role in developmental timing of Caenorhabditis elegans in the early 1990s [11], [12]. Subsequent studies have shown that cellular factors necessary for miRNA biogenesis and many miRNAs are conserved in many organisms, suggesting the importance of miRNAs during developmental processes and evolutions [13]–[17].
miRNAs are a novel class of non-coding small RNAs which have been recognized as global regulators of gene expression that control the key cellular processes such as growth, development and apoptosis [9], [10]. A single miRNA can potentially regulate several hundreds of mRNAs forming a complex regulatory network that can act in a flexible manner for precise and rapid effects on protein translation and gene expression. Majority of the miRNAs are expressed in a cell- or tissue-specific manner and may contribute to the establishment and/or maintenance of cellular and/or tissue identity. It is estimated that several thousand human genes, up to about one-third of the mRNA transcriptome, are potential targets for regulation by miRNAs encoded in the genome [18]. The regulatory process occurs posttranscriptionally and involves miRNA interaction with a target site in the mRNA that has partial or complete complementarity to the miRNA. The regulatory effect of miRNAs on gene expression is a complex process involving both translational repression and accelerated mRNA turnover, each of which appears to occur by multiple mechanisms. Moreover, certain miRNAs are also capable of activating translation [19], [20]. Hence, miRNAs are related to diverse cellular processes and regarded as important components of the gene regulatory network.
Importance of an individual miRNA is reflected in the diseases that may arise upon the loss, mutation or dysfunction of specific miRNAs [21]–[23]. One study reported mutations in 5 of 42 sequenced miRNAs in 11 of 75 patients with chronic lymphocytic leukemia. Although the majority of these mutations were somatic, at least one was germline [23]. Another study showed that up-regulation of several miRNA genes was correlated with loss of their target gene transcript (KIT) in papillary thyroid carcinoma. In 5 of 10 such cases, this down expression was associated with germline single-nucleotide changes in the two recognition sequences in KIT for these miRNAs [22]. Recently, a series of papers presented conceptually related ideas linking the genetic variations and alterations of biogenesis and function of miRNAs to the increased risk of developing sixteen major human diseases. Significant role of miRNAs in the pathogenesis of many major human disorders has been proposed as part of disease phenocode concept [24]–[26]. These results suggest that germline changes in miRNAs and their target genes may have a profound effect not only on disease progression but also an individual's risk of developing disease.
Current studies, however, have focused primarily on miRNA role as posttranscriptional regulators of target mRNAs or at a much higher level on their cell biological processes and organismal roles [9], [10]. Loss- or gain-of-function studies often analyzed effects on mRNAs by expressing or suppressing specific miRNA in cells. As these experiments create non-physiological levels of miRNAs that may affect target mRNAs abnormally, accurate evaluation of the miRNA effects may require normal range of variations in miRNA expression under an endogenous condition. Additionally, because miRNAs can interact with their target genes directly and influence expressions of many other genes indirectly, the miRNAs may demonstrate correlations with their target genes as well as non-target genes. Therefore, merely measuring target gene expression may not be sufficient to gain understanding miRNA regulatory effects. A complimentary approach is to identify downstream genes that are tightly correlated with fluctuation of miRNA expression. Liu et al [27] has reported significant miRNA correlations with target genes as well as non-target genes by performing expression profiling analysis on 12 brain tumor biopsies. Subsequent experimental validations demonstrated a directional causal relationship from miRNAs to mRNAs. However, the small sample size and potential mutations in the samples restrained statistical power to detect weak correlations.
To fully examine the miRNA-mRNA correlations at whole genome scale, we measured both miRNA and mRNA transcriptional profiles in 90 human Epstein-Bar virus transformed lymphoblastoid cell lines. We performed pair-wise correlation coefficient analysis and identified strong correlations between the endogenous variations in the miRNA and mRNA expression. Gene Ontology (GO) analysis identified over-representation of these miRNA-correlated genes in several biological processes. These high-throughput expression data provides a valuable resource to examine global effects of miRNAs on gene expression and hence on complex traits.
Endogenous correlations between miRNA and mRNA expression
We performed pair-wise correlation coefficient analysis to evaluate potential correlations between 366 miRNA and 14,174 mRNA expression levels. When false discovery rate q value (qFDR)<0.01 (approximately p value<0.00076), we detected significant correlation in 7,207 miRNA-mRNA pairs (Table S1), which were involved in 2,448 (17.27%) of the 14,174 mRNA probes and 90 (24.59%) of the 366 miRNAs. Of the 7,207 pairs, 4,085 and 3,122 pairs showed positive and negative correlations, respectively (Table 1, also see Table S2 for all 366*14,174 correlation coefficient r values). The most frequently involved miRNA was miR-363, which was correlated with 672 mRNAs. Cumulative frequency of these correlated genes for each of 366 miRNAs is shown in Figure 1A. Significance level of each mRNA probe for its correlation with miR-363 is demonstrated in Figure 1B.
A. 90 of the 366 individual miRNAs are correlated with at least one mRNA. We align the 90 miRNAs on 20 of 24 chromosomes based on their genomic locations (X axis). For each miRNA, the number of the correlated mRNA probes is demonstrated on Y axis (red dot). For example, miR-363 on Xq26.2 is correlated with 672 mRNA probes. Both miR-181b (correlated with 468 mRNA probes) and miR-181a (correlated with 440 mRNA probes) have two copies, each on different chromosomes (1 and 9). B. Among 11,417 known genes (14,174 mRNA probes), we successfully map 11,278 individual genes on the 24 chromosomes (X axis). We plot–log10 values of the correlation coefficient qFDRs between miR-363 and the 11,278 genes along Y axis (blue dot). The horizontal dot line (black) indicates qFDR = 0.01. Above the line are the mRNA probes that were significantly correlated with the miR-363. For example, the miR-363 is significantly associated with the gene SASH (chromosome 6q24.3) at qFDR = 3.70×10−11 (equal to 10.43 of −log scale in the figure).
When examining each of 7,207 significant miRNA-mRNA pairs individually, we found that positive correlations dominated highly correlated pairs. The positive correlations accounted for top 110 pairs (ranked based on qFDR values). The correlation between miR-10a and HOXB4 was the most significant with a positive correlation qFDR = 2.21×10−19. The most significant negative correlation was between miR-98 and SERBP1 (qFDR = 3.97×10−7), which was ranked 111th of the most significant pairs. Table 2 lists top 20 most significant positive and top 20 negative correlation pairs.
To confirm and validate the Illumina BeadArray data, we tested 6 miRNAs (miR-10a, -20b, -181b, -181c, -34b, -372) and 7 mRNA genes (GRK5, KIF3B, ADD3, HOXB4, SERBP1, ST7 and ZNF532) using TaqMan-based quantitative RT-PCR in each of the 90 lymphoblastoid cell lines. The 6 miRNAs were chosen because they demonstrated various degrees of correlations with mRNAs and were in the same multiplex RT pool (human pool 1) of TaqMan miRNA assays. The 7 mRNA genes were selected based on various levels of correlations with the selected miRNAs. Four of the six miRNAs (miR-10a, -20b, -181b, -181c) and all 7 mRNA genes had a Ct (cycle threshold) value≤35 and so were used in our final analysis. The results showed high level of concordance between the BeadArray and quantitative RT-PCR in 26 of the 28 comparisons (Table 3). Two miRNA-mRNA pairs (miR-20b and ST7, and miR-181c and ST7) gave poor reproducibility, and this may have resulted from the use of the two separate assays in targeting different isoforms (a and b) of the gene ST7.
miRNA-correlated genes and biological processes
To functionally classify miRNA-correlated genes, we used 11,417 known genes (14,174 mRNA probes) as a reference set and applied GOMiner (http://discover.nci.nih.gov/gominer/) for enrichment analysis of selected gene sets. We first evaluated GO classification for each miRNA-correlated gene set. To correct for multiple comparisons, we performed 1000 randomization analyses. For each of these gene sets, we observed various degrees of GO term enrichments. For example, eight of the top 20 miRNAs in Table 1 had at least one GO term that was significantly over-represented (randomization-corrected FDR<0.01). The most striking findings were from gene sets that were correlated with miR-331, miR-98 and miR-33b. For miR-331-correlated genes (269 genes from 284 probes), we detected significant over-representation for the GO terms of DNA replication (p = 2.65×10−23, 8.67 fold enrichment), DNA metabolic process (p = 2.35×10−22, 4.15 fold enrichment) and cell cycle (p = 1.94×10−19, 3.66 fold enrichment). Further analysis demonstrated that these enrichments were exclusively derived from negatively correlated genes. While we did not see any significant GO term in positively correlated genes, we observed significant enrichments in 58 GO terms for 191 negatively correlated genes (200 probes). Again, the most marked GO term included DNA replication (p = 9.21×10−28, 11.59 fold enrichment), cell cycle (p = 7.99×10−27, 4.89 fold enrichment) and DNA metabolic process (p = 1.09×10−26, 5.34 fold enrichment) (Table 4).
For the gene sets that were correlated with miR-98 (239 genes from 348 probes) and miR-33b (124 genes from 126 probes), we observed similar GO term enrichments as the miR-331 correlated genes. This was particularly true for the GO terms related to cell cycle. When considering both types of the correlated genes separately, however, we found the cell cycle-related GO term enrichments only in the miR-98 negatively correlated (153 genes from 159 probes) and in the miR-33b positively correlated gene sets (84 genes from 85 probes) (Table 4).
We then evaluated overall influence of miRNA expression on the GO categories. For a total of 2,248 miRNA-correlated genes (from 2,448 mRNA probes), we detected significant enrichments on several GO terms, in particular those pertinent to cell cycle with 72% (8 of 11 significantly enriched terms) relevant. Other significant GO terms included those related to cell communication, signal transduction and response to stimulus. To see if these cell cycle-related enrichments were driven by the miR-331, miR-98 and miR-33b, we excluded genes that were correlated with these 3 miRNAs and re-evaluated the GO term distribution. We found that cell cycle-related terms were no longer over-represented, but the two terms (cell communication and signal transduction) still remained enriched. Interestingly, further analysis identified that only positively correlated gene set contributed to the enrichments with p = 6.85×10−9 for cell communication and p = 1.05×10−7 for signal transduction (Table 4). Other enriched terms included immune system process (p = 6.30×10−7) and multicellular organismal process (p = 5.37×10−6). Table S3 provides these significant GO terms with randomization-corrected FDR<0.01 in detail.
Direct vs. indirect miRNA-mRNA correlations
To test if miRNA-correlated mRNA genes are direct miRNA targets, we downloaded the predicted miRNA targets from TargetScan5.1 (http://www.targetscan.org) and compared them with the miRNA-correlated genes. Because miRNA annotation file was based miRBase v9.1, some of miRNA names did not match the new version of TargetScan such as missing the -3p or -5p. Therefore, we limited our analysis to these miRNAs with name or sequence match in the TargetScan 5.1. We also limited the analysis to these genes with a reference sequence accession number. Before statistical analysis, we further filtered out the miRNAs that had less than 10 correlated mRNAs. Finally, 31 miRNAs were left for target prediction. We then compared each list of miRNA-correlated gene set to the list of predicted miRNA target genes. For the 31 miRNAs, we found 8 miRNAs whose targets were significantly enriched among their correlated gene sets (p<0.05 when included both conserved and non-conserved targets). The target gene set of miR-181a remained significant after multiple testing correction (FDR = 0.048).
Physical proximity and expression correlations
To estimate the effect of miRNA-mRNA proximity on their expression correlation, we extracted all miRNA-mRNA pairs that mapped on the same chromosomes and had correlation qFDR<0.01. We plotted r value of each correlation against distance between the miRNA and mRNA (Figure 2). For positive correlations, we observed clear trend of r value decrease when the distance increased. Specifically, the highest positive correlations were observed when miRNA and its corresponding mRNA was physically close each other on a chromosome. The highly positive correlations gradually decreased to baseline (at ∼≥2 Mb). For negative correlations, however, we observed constant r values from 0.4 Mb to over 140 Mb. We did not observe any correlation when the distance was <0.4 Mb, where 26 positive correlations existed.
We extract all miRNA-mRNA pairs that map on same chromosomes and have correlation qFDR<0.01. We plot the physical distances between miRNAs and mRNAs on x-axis and correlation coefficient r values on y-axis. The distance is based on the starting position of each miRNA and mRNA. We draw a logarithmic trend line (red) for positive and negative correlations separately. A. the distances from near 0 to 140 Mb are shown. For positive correlations (black dots), the trend line shows significant r value drop at far left side (short distance). However, for negative correlations (blue dots), the trend line tends to be constant. B. the distance from 0 to 0.5 Mb was shown. The positive correlations (26 pairs) are clearly clustered when the distance is <0.4 Mb (particularly <0.1 Mb), where no negative correlation is found.
miRNAs play an important role in regulation of gene expression. In this study, we examined genome-wide expression profiling in normal lymphoblastoid cell lines under routine culture condition and identified strong correlations between miRNA and mRNA expressions. Although complex gene-gene interactions may greatly diminish the power to identify significant miRNA effects, we were able to detect a variety of biological processes that may indicate function of those miRNAs. These results demonstrate that genome-wide transcriptional profiling analysis was able to detect endogenous correlations between miRNA and mRNA and that miRNA regulatory effect was discernable under natural culture condition.
miRNAs have been shown to target transcripts that encode proteins involved in cell cycle progression and cellular proliferation. Some of them display defective expression patterns in human tumors with the consequent alteration of target oncogene or tumor suppressor genes. These miRNAs modulate the major proliferation pathways through direct interaction with critical regulators such as MYC, RAS, PI3K/PTEN or ABL, as well as members of the retinoblastoma pathway, Cyclin-CDK complexes or cell cycle inhibitors of the INK4 or Cip/Kip families[28], [29]. It is postulated that by regulating an entire cellular program through the cooperative repression of target genes, miRNAs may serve as buffers to limit the accumulation of many gene products that impact cell cycle progression under a variety of contexts[28].
Interestingly, the miR-98 in this study is one of miRNAs that show significant association with cell cycle-related genes. The miR-98 is a member of let-7 family which plays a critical role in cell cycle control with respect to differentiation and tumorigenesis. The let-7 family is a master regulator of cell proliferation pathways by regulating the expression of the RAS as well as MYC oncogenes[30]–[32]. Over-expression of let-7 miRs alters cell cycle progression and reduces cell division in lung cancer cells[30] and causes cell cycle arrest by directly regulating the gene Cdc34 in human fibroblasts[33]. Clearly, the let-7 is a negative regulator of cell cycle process, which is consistent with our observation that miR-98 is negatively correlated with cell cycle genes in this study. Additionally, we also noticed a correlation of other let-7 members (especially let-7i and let-7g) with cell cycle-related genes (Table S1) in the lymphoblastoid cell lines, suggesting that let-7 family members do regulate cell cycle not only in fibroblasts and cancer cells but also in lymphoblastoid cell lines.
Because direct regulation of gene expression by miRNAs, we expect to see enrichment of miRNA-correlated genes among predicted targets. Indeed, we observed significant concordance between miRNA-correlated genes and miRNA predicted target genes in 8 of 31 miRNA sets. In particular, the miR-181a gene set showed significance of concordance even after multiple testing correction. For some reasons, we did not see any evidence of concordance in most miRNA sets. Generally, miRNA is believed to bind 3′UTR of a target gene and regulates gene expression at protein level. Therefore, miRNA target itself may not demonstrate noticeable change at mRNA level although some exceptions have been reported[34]. It is especially true when these correlations are examined under endogenous condition and there is limited range of variations in miRNA expression (contrary to extremely high level of expression by transfection study). Because miRNA inhibitory effect is at posttranscriptional level, the most significant changes could be downstream genes of the miRNA target at transcript level. Furthermore, because background noise and weak correlation between miRNAs and their targets, each tested miRNA had relatively small number of correlated genes when applying FDR<0.01. This also limited statistical power to detect significance.
Although the results from this study need further validation, the importance of the present study is clear. First, the study adopts a whole genome approach and identifies thousands of highly correlated miRNA-mRNA pairs. Second, the study measures expression levels under endogenous condition and without extremely high or low levels of miRNAs caused by transfection approach. Third, the study correlates miRNAs with their targets as well as downstream genes. The downstream genes are closer to the final consequences of miRNA-mRNA interactions. Fourth, the study identifies several biological processes that are associated with variations in miRNA expression. Lastly, the study results are supported by other publications using different materials and methods, demonstrating feasibility of this approach in studying miRNA function. We believe that these results along with supplementary data sets will provide a valuable resource to further investigate the miRNAs and their functions.
Materials and Methods
Ethics Statement
All subjects provided written informed consent; and the study was approved by the Mayo Clinic IRB.
Cell lines and Cell culture
We collected peripheral bloods from 90 Caucasian men with median age of 68 years old (44–74) and transformed the peripheral blood lymphocytes with Epstein-Bar virus to establish immortal cell lines. We then grew all transformed cell lines in RPMI 1640 media supplemented with 15% fetal bovine serum, and 1% penicillin/streptomycin at 37°C in humidified incubators in an atmosphere of 5% CO2. Experimental series were set up by seeding 5-ml cultures in T25 flasks. Each culture was fed with 5 ml of fresh media twice a week until the cell number reached ∼106 in a T75 flask. The cells were harvested and suspended in 500 µl of RNAlater and stored at −80°C for further processing.
RNA extraction
We extracted total RNA from each cell culture using miRNeasy Mini Kit (QIAGEN) under the manufacturer's guidelines. This protocol could effectively recover both mRNA and miRNA. The integrity of these total RNAs was assessed using an Agilent 2100 Bioanalyzer.
mRNA and miRNA microarrays
We used the Illumina human-6 V2 BeadChip for mRNAs profiling and Illumina microRNA expression profiling panel (based on miRbase release 9.0) for miRNA analysis according to the manufacturer's recommendation (Illumina, Inc., San Diego, CA). 200 ng of RNA from each cell culture was first labeled and then hybridized to each array using standard Illumina protocols. BeadChips (mRNA) or sample array matrices (miRNA) were scanned on an Illumina BeadArray reader. For mRNA, we repeated 30 samples in triplicate, 30 samples in duplicate and 30 singleton samples for a total of 180 expression profiles. For miRNA, we repeated 84 samples in duplicate and 6 samples in quadruplicate for a total of 192 expression profiles. We also arranged each of these replicates in separate arrays to reduce potential batch effect. Before data processing, we used various bioinformatics tools to examine the quality and reproducibility of each expression profiles. Based on principal component analysis, we removed 26 individual miRNA profiles due to substantial shifts away from a main cluster. However, replicates from each of the 26 individuals were still included in the analysis because they were in the main cluster. The expression profiles have been deposited in NCBI's Gene Expression Omnibus (GEO) with accession number GSE14794.
Data processing
We processed 180 mRNA and 166 miRNA profiles which included all 90 subjects. For both mRNA and miRNA data, we first transformed raw data generated from BeadStudio (Illumina, San Diego, CA) using a variance stabilization transformation algorithm [35] and then normalized them using quantile normalization implemented in Bioconductor (www.bioconductor.org). After normalization, the samples with replicates were averaged. As a large fraction of mRNAs and miRNAs were either not expressed or non-detectable, we filtered out probes with median detection p value≥0.01 (the p values were automatically reported in BeadStudio). This procedure reduced the number of mRNA probes from 48702 to 14174 and of miRNA probes from 736 to 366 for final data analysis. Among the 366 miRNAs, 273 are in miRBase database (http://microrna.sanger.ac.uk, v9.1), and 93 are potential miRNAs identified in a RAKE analysis[36], [37].
Data analysis
For each of the 366 miRNAs, we correlated them with 14,174 mRNA probes in the 90 individuals using Spearman's rank correlation analyses. We assessed statistical significance using q value of false discovery rate (qFDR) based on Storey et al [38] and Bonferroni correction. In this study, we defined the miRNA-mRNA correlation coefficient qFDR<0.01 to be statistically significant. We also reported our findings using Bonferroni-corrected p<0.05 (an equivalent to un-corrected p<9.64×10−9, 0.05 divided by 366×14,174). These analyses were done using Partek Genomics Suite (Partek Inc, St. Louis, Missouri). To explore whether miRNA affects expression of genes that share a common biological relationship, we searched for over-representation in GO categories from miRNA-correlated genes. We labeled genes with positive correlation (+1) and genes with negative correlation (−1). One gene list for each miRNA was submitted to the High-Throughput GoMiner at http://discover.nci.nih.gov/gominer/. The reference gene list was 11,417 known gene names from the 14,174 mRNA probes. The GO annotation was obtained by matching to the gene names in the UniProt database. We specified 1,000 randomizations for calculating the GO enrichment FDR q-value. The enrichment p-value for each GO category was calculated using Fisher exact test and the q-value was calculated using the distribution of the p-values obtained by randomly re-sampling from the reference genes[39], [40].
Real-Time PCR analyses of mRNAs and miRNAs
For mRNAs, 1.2 ug of total RNAs from each of the 90 subjects was converted to cDNA by High Capacity RNA-cDNA Master Mix (Applied Biosystems, Foster City, CA) in a 40 ul reaction according to the manufacturer's protocol. 1 ul of the cDNA was used for real time quantitative PCR in a total volume of 15 ul containing 1× TaqMan Gene Expression Master Mix and gene specific assay for selected genes which included GRK5 (Hs00992173), KIF3B (Hs01122781_m1), ADD3 (Hs00249895_m1), HOXB4 (Hs00256884_m1), SERPB1 (Hs00854675_gH), ST7 (Hs00251157_m1), ZNF532 (Hs00539543_m1) and GADPH (Applied Biosystems). For miRNAs, 0.4 ug of total RNAs was converted to cDNA using TaqMan MicroRNA Reverse Transcription Kit and multiplex RT primer pool 1. 1.8 ul of 1∶10 diluted cDNAs was used for real time quantitative PCR in a total volume of 15 ul containing 1× TaqMan Universal PCR Master Mix and specific assay for selected miRNAs which included miR-10a, miR-20b, miR-181b, miR181c, miR-34b, miR-372 and RUN48 (Applied Biosystems). All PCR assays were run in triplicate, and expression values were averaged. In order to increase the reliability, we excluded the assays with Ct value≥35, which included the miRNA assays for miR-34b and miR-372. To calculate the relative expression for each transcript, all real-time PCR data were normalized to GAPDH (mRNA) or RUN48 (miRNA) by ΔCt method. We also converted the ΔCt into a value of 20-ΔCt such that the latter value was positively proportional to the log of copy number and was comparable with log transformed data from microarrays.
Supporting Information
Table S1.
Excel spreadsheet containing all miRNA-mRNA pairs with significant correlation (qFDR<0.01).
Table S2.
Pairwise correlation coefficient r values between 366 miRNAs and 14,174 mRNAs.
Table S3.
Excel spreadsheet listing gene ontology analysis of miRNA-correlated genes.
We thank Mayo Clinic Advanced Genomics Technology Center for its excellent support on cell culture and microarray service.
Author Contributions
Conceived and designed the experiments: LW JC ST. Performed the experiments: LW JMC. Analyzed the data: LW ALO YWA HS. Contributed reagents/materials/analysis tools: LW SKM SMR ST. Wrote the paper: LW WL CJS SS JC ST.
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- 28. Carleton M, Cleary MA, Linsley PS (2007) MicroRNAs and cell cycle regulation. Cell Cycle 6: 2127–2132.M. CarletonMA ClearyPS Linsley2007MicroRNAs and cell cycle regulation.Cell Cycle621272132
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For example, say an HR department sends out an e-mail to all 1,000 employees announcing the company picnic and attaching a 1 megabyte (MB) jpeg map of the location. Depending on the e-mail system, when this e-mail and attachment is saved by employees, you potentially could have tens to hundreds of copies if this 1 MB e-mail consuming hundreds of MBs of storage. A CAS storage device would ensure only one copy of the e-mail is saved.
Drill down into today's new backup approaches
Due to the features of most modern backup software – which include snapshot management, DR elements, cloud support, VM protection – backups can do so much more than simply restoring data in the event of a storage or server failure. Download this guide and not only discover the latest upgrades to today's top enterprise backup vendors, but also learn where backup software works best in your computing environment.
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Posted by Lorie
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One brownie at a time
Darn Brownies was created for one reason — to give away brownies. We are roaming from city to city, baking brownies, setting up a card table and then handing out free dessert to everyone. There are no tricks. You can take the brownie and run. Our main goal is just to give away a snack and help people have a better day. But for those feeling generous, we do ask that you think about joining the brownie town improvement project. Read below and click through our pages to learn how we are trying to save the world one brownie at a time.
Darn Brownies –
Doing Actions that basically Result in Nothing
Click to read about our strict 50% zero tolerance policy on free brownies.
Join us
Send in a little project for your town, like giving away cake, and get entered in drawings for the official free brownies T-shirt. Or just tell us you support the movement and might come up with a foolish idea in the near future. This will make you a member as well as give you access to the site's signature recipe for Belgian chocolate chunk brownies.
Live brownies
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Send an e-mail to darngoodbrownies(at)gmail(dot)com to sign up for the blog or to receive updates on how one wooden brownie spatula is trying to change the world.
Mom's old-fashioned brownies recipe is available now on this site.
Join the movement to do something good for no good reason and receive the Belgian chocolate chunk brownies recipe.


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Another Cartoon Cavalcade episode hardly had the Old Trafford viewers laughing as United meekly ended their Champions League group section with a miserable defeat.
It may not have been as comic as Reading last Saturday but the Reds presented their opponents with a lead for the 14th time this season with some slapdash play and there seemed little urgency or desire to conjure up another of their famous comebacks.
As an exercise in proving United have an underbelly that can carry them all the way to Wembley in May when Europe's elite stand in their way this was totally unconvincing.
The flaws from last season's group exit and the Europa League failings were all too evident again.
Click here for the match centre to see how the action unfolded and for stats and standings
Reds boss Sir Alex Ferguson believes the group saunter that had seen United top Group H and qualify early underlined the progress his squad were making.
However, this was a huge backward step. Never mind Real Madrid and Milan relishing being pulled out of the hat with United on December 20 for the knockout first stage, on this evidence Celtic will also be fancying their chances.
Twelve months ago five of the starters in this dead rubber were experiencing the final miserable throes of the 2011 Champions League fiasco.
David de Gea, Wayne Rooney, Ryan Giggs, Chris Smalling and Phil Jones all suffered the ignominy of Basel dumping the Reds out in Switzerland.
Danny Welbeck came on as a sub in a bid to rescue United's embarrassment but it was all to no avail.
That chastening experience has galvanised the Reds into a better European outfit this term – even if it has been steady rather than spectacular progress to the knockout stages.
So for that sextet compared to the trauma of a year ago this was a much less tense affair.
The downside, however, for arguably all but Wayne Rooney was that they knew pulling on the shirt against the Romanians almost certainly meant they wouldn't be doing the same come kick-off at the Etihad on Sunday.
Scott Wootton is way down the pecking order behind the established rearguard and back- ups like Smalling and Jones, yet he looked the steadiest of United's defence as Cluj went about the business of attempting to sneak into the last 16 via the back door.
The last senior outing for 21-year-old Wootton was at Stamford Bridge in the Capital One Cup. It was a toil against Chelsea in London but, thankfully, it hasn't inflicted any lasting damage.
The first major chance of the first-half saw Modou Sougou put through far too easily.
But Wootton spotted the danger and slid in to make a vital block as the Senegalese striker pulled the trigger.
He was then first to read the danger to cut out a dangerous low cross from Camora.
Also alert defensively was Rooney.
Fergie had made the decision to keep his striker active following a return to form with two goals amid the Reading mayhem.
As well as his goals at the Madejski, the England striker's work-rate was excellent as he helped out the comedic defence.
Again last night Rooney had to trawl back to aid the suspect rearguard, blocking two efforts.
City will have been happy to see him doing the draining work and as a goal threat you wondered if this outing was worth the effort.
But in the 41st minute he turned the heat up, clipping a delightful chip that almost caught out keeper Mario Felgueiras.
Cluj's number one did well to flip the effort over the bar and from Rooney's corner Smalling glanced a header wide that he will watch again and again and wonder how he didn't score.
Welbeck was asking the same a couple of minutes earlier when his glancing header was kept out by a fine reaction save from Felgueiras.
That late first-half spurt suggested United would wake up in the second period but they went down the gears rather than up.
It obviously doesn't matter whatever mixture of names Fergie writes on his team-sheet – the flaw in the make-up of this current squad is the giveaway goal that puts the Reds on the rack.
Cluj's danger had been minimal but in the 56th minute Paul Scholes, on as a sub for the injured Tom Cleverley, played an errant ball and the Romanians surged forward.
Brazilian Luis Alberto unleashed a South American cracker that swerved past a helpless David de Gea.
For the 14th time this season United had ridiculously given themselves the job of having to conjure up a rescue act.
Normally at this point one would see Fergie prowling the technical area, urging his troops to respond.
But the Reds manager clearly had succumbed to the occasion and preferred to stay in the VIP seats alongside chief executive David Gill.
It had obviously been a tough sell attempting to shift tickets for this non-event.
When the PA announcer read out a 71,521 crowd the stadium fell about. That many may have left the ticket office but many mustn't have bothered using them.
They were perhaps the fortunate ones and good judges.
The hotter tickets will come in February when the real business starts in Europe.
By then, it's to be hoped someone will have found a solution to stopping the Reds pressing the self-destruct button.
What is your verdict on the performance? Have your say.
Manchester United 0 CFR Cluj 1: Stuart Mathieson's Champions League match report
Another Cartoon Cavalcade episode hardly had the Old Trafford viewers laughing as United meekly ended their Champions League group section with a miserable defeat.
Another Cartoon Cavalcade episode hardly had the Old Trafford viewers laughing as United meekly ended their Champions League group section with a miserable defeat.


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MMA Forum - UFC Forums - UFC Results - MMA Videos (http://www.mmaforum.com/forumindex.php)
- UFC (http://www.mmaforum.com/ufc/)
- - 'It Will Be A Stand Up War.' - Gustaf. (http://www.mmaforum.com/ufc/107255-will-stand-up-war-gustaf.html)
'It Will Be A Stand Up War.' - Gustaf.
I believe Gustaf is referring to Rua's boxing when saying he lacks proper technique, which is true. But if anyone can make this fight ugly and knock Gus back, it's Shogun.
Really wished they didn't put this fight together for obvious reasons. I'm a fan of Shoguns, but I want Gustaf to fight the LHW title holder and for Shogun to get his rematch. If Shogun loses, man it'll be a long time before he gets his opportunity again.
Fight wise, no doubt it's going to be explosive. It would appear the momentum is on Gustaf's side + speed. Very interesting fight.
"...Shogun is his heart. He is willing to take a beating to get what he wants, so it's a fighter you have to - the more you punch him the better he gets..." So true
If Shogun wins then we have nothing much to hope in the light heavyweight division. If Gustafsson wins we have a new intriguing matchup in Jones vs Gustafsson. Guess which one I am cheering for ?
btw: I am Swedish as well :thumb02:
Shogun won't ever fight Jones again even if he gets to where he deserves a shot he won't take the fight.
Sent from my Samsung Galaxy S3
I think it's more of redemption than anything. The LHW title holder made one of THE TOUGHEST SOBS tap out. That whole fight was pretty tough to digest. Then he chokes out Lyoto.
May the best fighter win.
- Gustaf
- Glover
- Mousassi
- now that GSP has pretty much declined the super fight perhaps the Greatest will move up north once again.
There's actually some potential matchups out there in the division.
Might be wise for Shogun if he tries to turn this into a grappling match. Not like Gustafs TDD is all that spectacular. He might have better technical boxing but Shogun is much more well-rounded and has a ton more experience. He needs to make it a grinder to win.
Anderson Silva and Machida are the most probably guys to beat Jones. If Gustafsson wins he is going to be included as well. What is disheartening though is that Jones slices through all opponent like knife through butter. Jones beat Rampage and Shogun compared to how Machida is 1-1 and once knocked out by Shogun plus lost a close fight to Rampage. That is overwhelming stats in favour of Jones and despite styles makes a fight Jones is still a major favourite likely to win.
Anderson Silva has the best shot because even if he gets taken down he still has a chance. He can also outmeneuver Jones standing up despite that range disadvantage.
Gustafsson offers more of quick knockout chance and given his footwork, speed and tallness it is very hard to stay away from him. I rate Gustafssons chances vs Jones higher than that of Machidas.
I hope this fight gets dirty, I want to see Alex's chin and heart. If he is able to overcome Shoguns assault, he may have a prayer against Bones.
I really want to see Gustaf fight Jones. Jones is beating great fighters but he has not fought a young lion like himself, with size to match, since Bader. And Bader is hardly a lion.
Shogun is on the decline but he is a born-to-fight fighter. Meaning no matter what shape he's in, he's a handful. Unlikely to hold the belt again, I don't know how much he wants it, but he can certainly put other guys' dreams on pause.
I really want to include Gustaf in my betting but he's going to bang with Shogun... I can't risk a parlay on that.
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From Iris' Catster Photos[ View profile page | Plus Photobook ]
|"Thank you Tundra, Rory, Manytoes, & Lynzee!"
You're welcome, Iris!
Your cat loves a clean toilet -- don't we all? Catster's singing Cat Lady, Sarah Donner, explains why.24
|"Thank you Tundra, Rory, Manytoes, & Lynzee!"
November 15th 2012 at 10:09 am
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Wednesday, May 7, 2008
Collective Consciousness
That's exactly how I am feeling about A New Earth, by Eckhart Tolle. I feel like he is giving me new lenses to see the world through. Not like my old way of seeing the world was wrong, but I felt a little nudge within me to discover things a bit deeper in my life. Of course the book kept coming up in conversation and I thought, I may as well give it a read. Honestly, I had nothing to lose.
Turns out just in the first four chapters, I have started to discover what I have to gain. Reading this book, grounded me in the reality that life is a journey. My path is always changing and I can help steer it's course. I can learn to let go of notions and feelings that held their grip so tightly on me it affected my sleep, or my mood, or how I mother my girls, or how I love my husband, or how I nurture my friendships.
Right now, I am embracing the opportunity to examine the negativity in my life, chronic complaining, and how to let things go. Working on my perspective, my attitude and seeing where the change leads me. Keeping track of my distractions and are they really fulfilling the emptiness I am feeling or creating more wanting?!
I firmly believe opportunities for growth can happen in your life when the timing is just right, even if you didn't know it was the right time.
Last month, after picking up the book for a steal at Costco (and no bananas, may I add), I sat down and gave the book a quick glance. I decided I would see if I could gather a group of women to read the book with me. Oprah has fabulous resources with workbooks and Podcasts, so the work for the group was essentially done. I just needed to find a time, place, and those that were interested.
Much of the content of the book touches upon topics which can have such different meanings to the individuals gathered reading the book as a group. We all seem to open up in our way, learn things about ourselves as we are learning about one another. My hope it to have my hubby read the book. I know I will be re-reading the book, many times over. Book club will continue over the next month or so, as we are taking it slow.
Maybe you'll notice a new spring in my words, for Tolle's words are speaking to me loud and clearly.
8 comments:
Okay, I think I'm the only person I know who didn't love this book. I just couldn't get into it. I don't think I'm evolved enough for it yet. That whole body-pain thing, why some babies cry more, etc just went right over my head and I didn't buy into the theory.ReplyDelete
That said, I immensely enjoyed hearing him speak on the Oprah podcasts (although I only heard about 2 casts). Discussing it made it alot more digestible for me. I'm anxious to hear more of your thoughts.
One of the aestheticians at work has that book on her desk. I see it every time I go in to her treatment room. I keep thinking about picking it up but haven't gotten around to doing so yet.ReplyDelete
I am so glad to hear the book is helping you. Granted, you don't have to take everything so seriously; but I think it's a great guide to putting your life into perspective.ReplyDelete
- AnonymousMay 7, 2008 at 7:15 PM
Thanks for including me in this journey my dear. I am so glad to hear that it is so inspirational for you. As I sit hear watching my mind react to my children climbing on the furniture, and the one who just hit their head on the table, the one who slammed the fridge door, and is now having a tantrum asking for more sugar. Love, KSReplyDelete
That book sounds good--I'll look for it at Costco. I love Costco for books--I basically just collect them, figuring I'll get around to reading them later. :)ReplyDelete
Have I cracked it since Sunday? Are you kidding? ;) I will, though. I'm ready for it. I *need* it.ReplyDelete
after my mood yesterday I REALLY need to read some more. :)ReplyDelete
I haven't gotten around to getting the book yet. However I am very curious as to what the big buzz is all about. Thanks for sharing your experience.ReplyDelete


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You gotta be tough to be a Cleveland Browns fan. Real tough.
What happened Thursday morning at the Browns' training camp was not for the faint-hearted or thin-skinned. It was a test, yet another in a long line of tests, to gauge just how impenetrable the fans' love is for their team.
When LeCharles Bentley was carted off the field with an injured knee on the first play of contact, it was as though the entire Browns Nation had been punched in the midsection.
Feel familiar? It should.
Remember the Sports Illustrated cover when Art Modell pulled the plug on Cleveland back in 1995? It was a feeling SI captured perfectly. Happened – to a lesser degree, of course – again yesterday.
The news that the Browns' best offensive lineman went down triggered a psychological bender on this Web site. Teeth gnashed, psyches were broken, tears were shed, regurgitation scattered the landscape, spirits were shattered, profanities proliferated.
And who could blame the fans?
Emotions flowed. From anger to frustration to paranoia to bewilderment to disbelief to shock. Laugh to keep from crying. Punch a wall instead of another human being. Kick the waste basket instead of the pet.
What better way to vent.
Only in Cleveland . . . the Browns are jinxed, cursed . . . who's next ? . . . goodbye 2006 . . . what time do the bars open?
Others, perhaps rationalizing, took it a lot better. "What else should we expect?" "This kind of crap happens all the time. We all should be used to it by now."
The hurt was palpable from coast to coast.
The good feeling that always accompanies the opening of a training camp disappeared in a heap. And it certainly caused one to wonder – again – exactly what did Cleveland do to deserve all this?
Just when it looked as though the Browns had turned a corner by signing the likes of homeboys Joe Jurevicius, Dave Zastudil, Bob Hallen and Bentley, something like this happens. And on the second day of camp, no less.
Bentley is no ordinary player. He's not just the St. Ignatius kid living a dream of playing for his hometown team.
He is a Pro Bowler, an honor that is somewhat foreign to Browns players. He was going to help solve a lot of problems along the offensive line.
He was clearly the best player – since Jamir Miller – the Browns have had since their reincarnation in 1999. He was going to be the anchor of the line, the linchpin, the man around whom the team would rally.
Bentley brought an infectious glow to the locker room. Always has a smile on his face. The fact he was now a Brown surely would keep that smile permanent.
So why did this happen? Why, why, why?
There are no good answers to why this happens to the Browns, whose history with significant injuries to key players is almost legendary.
As much as we want to believe there is a jinx, a curse that dangles over the team, rational thought suggests otherwise. It's just a little difficult at this time to fight through those initial thoughts and see the bigger picture.
Some posters suggest that God hates the Browns. No He doesn't. If He didn't think you could handle it, He wouldn't keep doing this to you.
Somehow, some way, Browns fans always come out of the doom and gloom and see the brightness. Why? Because they're tough, resilient.
Otherwise, they wouldn't be Browns fans.
Passan: Another Test of Faith
You gotta be tough to be a Cleveland Browns fan. Real tough.


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THE SECRET SISTERS JOIN IRON & WINE, SHE & HIM FOR SUMMER TOUR New Single "Let There Be Lonely" Available Digitally Featured on ABC's Nashville March 27
Alabama's THE SECRET SISTERS make their return to the road touring with Iron & Wine then She & Him in June on tour. The jaunt includes a performance at the legendary Ryman Auditorium in Nashville, TN on June 13. They remain one of the genre's most beloved live acts, and they'll be enchanting audiences throughout 2013 beginning with these summer dates.
The girls released "Let There Be Lonely" from their highly anticipated sophomore album produced by legendary producer T Bone Burnett for Republic Records on March 19th via iTunes and all digital retailers. It's a soulful slice of Americana, incorporating folk, roots, and country, and it's carried by their passionate and powerful vocals harmonies. It's punctuated by all the hallmarks of their sound that fans have come to grow and love.
In addition, the song will also figure prominently in an episode of ABC's hit drama, Nashville, airing March 27. Avery, played by Jonathan Jackson, will perform it during a key scene.
The Secret Sisters "Let There Be Lonely"
Share in the magic of THE SECRET SISTERS with their forthcoming new music and impending tour.
Confirmed Dates
3/27 Chapel Hill, NC Memorial Hall*
4/27 Huntsville, AL Panoply Festival***
5/12 Napa, CA Bottle Rock Festival***
5/13 Pittsburgh, PA Carnagie Music Hall of Homestead*
5/14 New York, NY Beacon Theater*
5/15 Philadelphia, PA Union Transfer*
5/16 Richmond, VA Lewis Ginter Botanical Gardens*
5/17 Boston, MA Berkelee Performance Center*
5/18 Portland, ME State Theater*
5/19 Port Chester, NY Capitol Theater*
6/13 Nashville, TN Ryman Auditorium**
6/15 Grand Prairie, TX Verizon Theatre @ Grand Prairie**
6/16 Austin, TX ACL Live at the Moody Theatre**
6/18 Phoenix, AZ Comerica Theatre**
*w/ Iron & Wine
**w/ She & Him
*** Festival Appearances
A division of Universal Music Group, the world's leading music company, Republic is home to an all-star roster of multi-platinum, award-winning legends and superstar artists such as 3 Doors Down, Amy Winehouse, Black Sabbath, Colbie Caillat, Eddie Vedder, Enrique Iglesias, Florence + the Machine, Godsmack, Owl City, The Voice (NBC-TV), Gotye, Of Monsters & Men, Mayer Hawthorne, among many others. It is comprised of innovative imprints and digital business ventures including Republic Nashville (The Band Perry), Cash Money (Lil Wayne, Drake, Nicki Minaj), Indie Pop (The Cataracs), Lava Records (Jessie J, Black Veil Brides), Aware Records (Mat Kearney), Brushfire (Jack Johnson, G. Love), XO (The Weeknd), Loma Vista (Soundgarden, Damian Marley) among others.


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Google are celebrating the 161st birthday of revered Spanish architect Antoni Gaudi in a new Google Doodle graphic.
Born in 1852 in the small town of Reus, Catalonia, Gaudi went on to become celebrated for his exuberant style of design, and ended up creating some of the most iconic attractions in Barcelona.
Today's Google Doodle tribute depicts Park Güell, Gaudi's famous garden complex in the north of Barcelona, the shell-like interior of the Casa Mila and the balcony designs of Casa Battlo, two of his most emblematic buildings, and the masked soldiers of his seminal, unfinished work, the Sagrada Familia.
The construction of the Sagrada Familia church was started in 1882 by architect Francesc de P. del Villar in a Gothic revivalist style, but when Gaudì took over he changed this, working on the building until his death.
- More about:
- Architecture
- Barcelona


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Courtesy: www.tomwolf.smugmug.com
Related Links
As part of the Breast Cancer Awareness Night, the Golden Griffins will auction off their gameworn pink uniforms to the highest bidder following the game. The event will also feature a raffle and bake sale. All proceeds raised will be donated to the Roswell Park Cancer Institute towards breast cancer awareness.
The event is free and open to the public. For more information, call (716) 888-8489.


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Coconut Chocolate Blondies
As far as sweets go, I tend to rely a little too much on a few certain items – chocolate chip cookies and brownies are on high rotation. I don't usually give much thought to blondies. But I've come across a few recipes lately, enough to get me thinking about them. So when I felt like baking this week they were at the top of my list. I used this recipe by the Brown Eyed Baker and it was awesome. The coconut and chocolate really give these guys an edge. The coconut in particular makes them seem almost tropical. It's perfectly summer-y twist on a classic. And, I know most of the continent is baking in a heat wave, but for my fellow North West Coasters who are still waiting (*sigh*) for the sun, give these babies a go. You'll be happy you did.
Om nom nom.
Food Links
A sampling of the yummy things from across the web that caught my eye this week:
The Savory
Warm chickpea salad with cumin and garlic { The Kitchn }
Yummy taco salad { Everybody Likes Sandwiches }
Beer battered cajun fries { The Kitchn }
Charred corn tacos with zucchini radish slaw { Smitten Kitchen }
Homemade pretzel rolls { Food Network }
The Sweet
Cake Pops { Oh My Sugar High }
Chocolate chip cookies { Real Simple via Cupcakes and Cashmere }
Whiskey peach and plum galette { Crepes of Wrath }
Carrot sheet cake with cream cheese frosting { The Kitchn }
Honey and sour cream cupcakes with whiskey icing { London Bakes }
Bon appetit!
If you are looking to give your space a dose of style- whether you are splashing out or on a budget – drop me a line. Check out my Hire Me! page for more details.


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Baker 33 MM Stainless Steel Women's Watch
$225.00 Actual transaction amount
Item ID: jb270415-003-01
Baker Bracelet 33MM Women's Watch by Marc by Marc Jacobs is modern everyday sporty chic. Features 33mm round gold tone stainless steel case, white dial, gold tone bracelet with deployant clasp, gold tone "Marc" with beveled sticks and hands and minute dial. Scratch-resistant sapphire crystal face, Japan Quartz Movement and water resistant to 5 ATM. Genuine Marc by Marc Jacobs.
Delivery & Returns
shipping & delivery info
This item is currently available upon request
This item is handcrafted upon request. Time indicated reflects our current waiting list.
This item can usually be crafted also according to your size request. Please contact our product specialists to find out more.


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I recently received, courtesy of the publisher, a copy of the new book The Anointed: Evangelical Truth in a Secular Age by Randall Stephens, an associate professor of history at Eastern Nazarene College and Karl Giberson, formerly a professor of Physics at Eastern Nazarene. Giberson has now moved on to concentrate on a number of [Read More…]
Anointed? … Evangelicals and Authority 1 (RJS)
Nov 3, 2011 @ 5:15 by 38 Comments


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Agenda – 2011/9/9
Campus Senate Meeting
Friday, September 11, 2009
10:00 a.m. – 12:00 p.m.
PAC Conference Room H
I. Approval of the Agenda
II. Approval of Minutes from meetings of August 28, 2009
III. Announcements
IV. Reports:
- Chair – T. Ting
- Provost – H. Berman
- Student Government Association – M. Van Vossen
- Committee on Committees – L. Fisher
- H1N1 Flu Update – Lynn Price
- Admissions Reforms Report – T. Barnett
Reform Response
Grad Admissions Table
Undergrad Admissions Table
V. Old Business:
- Resolution 39-2 Recognize Academic Honors at Commencement [2nd reading]
- Resolution 39-4 Changes to the UIS Campus Senate Bylaws Article VI. Section 2. and Section 2.C.2 [2nd reading]
VI. New Business:
- Resolution 39-5 Social Responsibility and Leadership Development Minor by EXS-L [1st reading]
UGC Recommendations memo
Dean's memo
CLAS memo
- Resolution 39-6 Changes to the UIS Campus Senate Bylaws – Intercollegiate Athletics Committee [1st reading]
- Resolution 39-7 Change to the UIS Campus Senate Bylaws – Committee on Diversity, Equal Right, Opportunity and Access [1st reading]
VII. Adjournment


text: Thats what I do with all my junk mail return envelopes. I find good articles that interest me and send it back to em. I don't know if the one who opens them up can read English or not or they just get thrown in the garbage. It's still worth a try and it's their dime.


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Saturday, November 27, 2010
Found on the Plains an Prairies of the West, but known in the East as well, they provided Food for the people. Now, with Potash, it is and will be global food for the people
The Multi-Mineral Supra-National Corporations are hunting. The Canadian Govt, in a spate of nationalism, quashed the potential take-over of Potash Corp by a MMSNC, but let POTASH One, get hitched to a German firm, K & S. Hmm? How many Juniors are out there now? Whom are worthy ? Who will be left ? And whom will make a run for it and who will stand and fight a lo-ball take-out?
As much as the heartburn hurt watching these Juniors languish, it was obvious in the rising tide of global food production, that a time of PEAK POTASH would rise.
Patience AND being Right paid off in a quad off the 52 week low, and a little bit less from its selection.
What I ponder now is, if our prime candidate in this stampede has more than a double left ? I think it does but have to wait for the next vignette to unfold next week.
www.denaliguide.us will carry Chat Links and DGS Network links to forums & Chat.
Subscribe to: Posts (Atom)


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LibraryThing Author:
Member: pooks
CollectionsYour library (321), Currently reading (1), All collections (325)
Reviews5 reviews
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About meAward-winning screenwriter and novelist Patricia Burroughs loves dogs, books, movies, and football. A lifelong Anglophile, she treasures her frequent travels in the British Isles researching current science fiction/fantasy writing projects. She and her high school sweetheart husband are living happily ever after in their hometown of Dallas, Texas.
Also onFacebook, Tumblr, Twitter, Wordpress
Membership LibraryThing Early Reviewers/Member Giveaway
Real namePatricia Burroughs
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Member sinceMay 3, 2007
Currently readingA Red Herring Without Mustard: A Flavia de Luce Novel (Flavia de Luce Mysteries) by Alan Bradley
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Maxx Shots Photographer
Miss FionaModel Female San Francisco, California, US
My MM URL: http://www.modelmayhem.com/missfiona
Mayhem # 372904
About Me
Hey guys! Thanks for looking at my port.… I am trying to find more time to pursue modeling! I am open for all types of projects, however I am not too interested in nude…I have modeled most of my life, and done a little acting.. I have also been on the Disney show in the UK, I am outgoing, fun and always enjoy having a great time on shoots. I love meeting new people and I also love to collaborate on many things!
I am looking for fun people to shoot with so I can start building my port again… it has been a long time so I am very egger to get back into it!
Send me a message if your interested!
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News and Rumors
An Alex Rios Trade to the Pirates Now Seems Unlikely
News and Rumors
Dave Kaplan of CSN Chicago says the Pirates and the White Sox sound close on a trade for Alex Rios.
Kaplan doesn't make it sound like a deal is a guarantee, since he says he's not sure if the two teams can come to a middle ground on the money. Rios is owed $18 M over the next year and two months. Earlier today we heard that the White Sox wanted Jameson Taillon or Gregory Polanco for Rios, but that the Pirates said no. We also heard the White Sox like Alex Dickerson.
UPDATE 1:47 PM: Thoughts from James Santelli on Rios...
Though Jose Tabata's return from injury improves the Pirates in right field, it is still a very bad position on the team. The Bucs are second-to-last in the National League in Wins Above Replacement (-1.0 ...
This content is for Pirates Prospects subscribers only. Subscribers get access to all of our daily articles on the Pirates and their minor league system, with live coverage throughout the system on a daily basis. Our lowest rates are $2.22 per month under our Top Prospect Plan, which also gets you a 40% discount on the 2017 Prospect Guide. Subscribe today to access all of our daily coverage of the Pirates' system.
If you're already a member, you can log in below. If you think you're receiving this message in error, please e-mail firstname.lastname@example.com.
Tim is the owner and editor in chief of Pirates Prospects. He started the site in January 2009, and turned it into his full time job during the 2011 season. Prior to starting Pirates Prospects, Tim worked with AccuScore.com, providing MLB, NHL, and NFL coverage to various national media outlets, including ESPN Insider, USA Today, Yahoo Sports, and the Wall Street Journal. He also writes the annual Prospect Guide, which is sold through the site. Tim lives in Bradenton, where he provides live coverage all year of Spring Training, mini camp, instructs, the Bradenton Marauders, and the GCL Pirates.


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Items in Cart:
Nokia N900
The Nokia N900 isn't so much a phone with internet capabilities as a handheld computer with phone capabilities. It runs on Maemo, a Linux based operating system supported by major players like Intel, and an improvement over Symbian. The phone has a clean design and the face no physical buttons, comprising almost entirely of the enormous screen. The QWERTY sliding keyboard which considerably increases typing comfort also makes the phone comfortingly chunky and substantial to hold.
The main features of the Nokia N900 are:
All in all, the Nokia N900 is by far the best of Nokia's touch-screen smart phone offerings, and delivers a truly desktop-like experience, thanks to its Maemo platform. Some of the N900's features like the camera and video recording are superior to the iPhone's, and it is probably the first smart phone to be something of a challenger to the iPhone 3GS. The Nokia N900's Internet capability is also superior to that of the Blackberry Storm 9530, although the email experience may be better on the Blackberry.


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White Girl Salsa Review
I am so excited to finally get this review up. It has been a long-time in the making but this summer was a bit crazy from a scheduling standpoint. So, let's file this under the better late than never category. The last Food Fête that I attended yielded many new gluten-free finds but there was a true standout among all the brands that were displaying their wares…
Hello, Old Friend.
It is no secret that I love to snack. I mean what goes better with the perfect cocktail than something wonderfully salty and savory Pretzels are the one snack food that has come a very long way since my celiac diagnosis many, many years ago. There was one product that was one of my all-time favorite snacks when they came out and I have never…
A very big congratulations to Cara from Texas who was selected as the winner in my One for Me, One for You gluten-free giveaway. Thirty six amazing gluten-free items that I hand curated will soon be making their way to the Lone Star State.
My One For Me, One For You, Ultimate Giveaway
Every year I like to do a giveaway as my personal thank you to my readers. I do this giveaway on my own dime and no items have been provided to me. Here's how it works…
My 2012 Gluten Free Year In Review
As 2012 winds down I thought it was time to wax nostalgic and take a look back at some of my favorite gluten-free moments of the previous twelve months.


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- 4 total posts
- 1 comment
- 3 questions
Questions from Stina
Any Advice on How I Should Prepare Myself for a Tummy Tuck?
I am so ecstatic about getting my tummy tuck done next week after having waited for a very long time. READ MORE
Should I be Able to Walk Straight 3 Days After Tummy Tuck Surgery?
I had the full tummy tuck w/ muscle repair and lipo on upper hips. My pain has been tolerable since day 1 post op and it's day 3 and I am hardly on any pain meds. My drain... READ MORE
Recent comments from Stina
Aww Happy Birthday in advance and Good luck on your future procedure. (= Iam also getting a tummy tuck done this coming Tuesday and am nervous and excited at the same time. READ COMMENT


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If you've ever wanted to look just like the vapid, gluttonous elite of a dystopian dictatorship that forces its children to murder each other both for mass entertainment and as a means of social control, you're in luck!
No, the Hunger Games Capital Colours nail polish collection isn't something from SNL: it's all too real. China Glaze, the company behind it, seems to have missed that the whole "Capitol look" was supposed to signify the corruption of the elite's souls, and not their kicky fashion sense. Then again, maybe "monstrous, inhuman idiot" is the new look for 2012. I've never been accused of fashion-forwardness.
I can't imagine a better look for There Will Be Blood Night at Brunswick Lanes, or hitting the links with the Funny Games golf-club collection. How about you?
(And if you want to talk about The Hunger Games, we've got a hopping discussion going on in our forums right now...)
There's even an official Tumblr dedicated to "Capitol Couture".
On one hand, this seems to miss the point?
OTOH, you get a lot more Vaders, Storm Troopers and bounty hunters at cons than you get Lukes, rebel troops or ewoks.
OTGH, nail polish? Really?
ironic isn't it? I think that the movie made the capital folks look too pretty...where were the people with dyed skin and tatoos all over their faces??
Similar sentiments with The Lorax and how it was marketed like disposable Popcorn bags, advertising an SUV to school kids, slapping "Lorax/Truffula tree approved/friendly" labels on cleaning products...
View All →


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Giving Carlos Beltran His PropsJuly 28, 2011
Based on all the published reports that Carlos Beltran has been traded to the San Francisco Giants, I decided to make a video giving Beltran the credit he deserves for the numbers he put in in the 6 1/2 years he was in a Mets uniform. Watch the video below and let me know your thoughts.
This entry was posted in New York Mets, On The Black TV and tagged Carlos Beltran. Bookmark the permalink.


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The program requires the completion of five courses, with a minimum grade of C-. Prematriculation credits may not be applied towards the requirements for the minor. Minimum grade policy applies to students entering September 2013 or later.
The Physics Minor Program requires completion of five courses.
Physics Minor*:
Five courses in physics, which must include Physics 13. The other courses may be selected from among Physics 11, 12 (or 1, 2), and courses in physics numbered 15 or higher.
Astrophysics Minor*:
Five courses, at least three of which must be courses in astronomy numbered 15 or higher. The remaining courses may be selected from among Physics 11, 12 (or 1, 2), 13, and courses in physics or astronomy numbered 15 or higher.


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Hey there, time traveller!
This article was published 27/7/2012 (1796 days ago), so information in it may no longer be current.
So I know Vine Lines is supposed to be about wine and not about books.
But there's a connection here, and I hope you'll bear with me until I make it.
Bottom line, though: "Let's Pretend This Never Happened (A Mostly True Memoir)" by Jenny Lawson is, without doubt, the funniest book I've ever read.
Anyone who either works in Human Resources or has worked in HR in the past should read this book. Anybody who suffers, as I do, from panic or anxiety disorders should read this book. Anyone who's married, or who has been married in the past, should read this book.
If you're directionally challenged, read the book. If you had a somewhat unorthodox upbringing, pick up a copy. If you went through a rebellious stage as you were growing up, for pity's sake, grab this volume and READ IT!
I'm a huge fan of Dave Barry, the former Miami Herald humour columnist. His books and his takes on everything, from the marvelous to the mundane, were always the most mirth-inducing writings I'd ever had the pleasure of reading.
Until now.
Lawson's hysterical, stream-of-consciousness, wrenchingly honest, and oh-so-believably real musings have had me in stitches since the first page, and they haven't let up. At all.
It's gotten so bad, my neighbours are starting to look at me oddly. Which I understand, because I've done most of my perusing of "Let's Pretend" on my deck. And I've gone off into gales of laughter — sometimes to the point where I shriek so much I can't breathe, so the combinations of sounds emanating from my deck are verging on other-worldly.
I can hear conversations in nearby yards become murmurs that quite quickly stop altogether, as they wonder what sort of weirdo is doing what sort of freakish thing a few doors down.
And I apologize. But I simply can't help myself. This stuff is so incredibly funny, I'm powerless to temper my reaction.
Now a brief caveat. Two, in fact. First, a person's sense of humour, just like a person's taste in wine, is completely individual. And what one finds funny, another might find ridiculous in a completely non-funny way.
Second, if you've a low tolerance for profanity, "Let's Pretend" is not likely the book for you. While I don't appreciate conversations that are laden with F-bombs, I've been exposed to enough of them that, while it still makes me shake my head in exasperation (because really — there are SO many other words out there to choose from), it doesn't bother me quite as much as it used to. And for reasons I can't explain, I find that particular word much more offensive spoken than I do in written form.
But as part of Lawson's vernacular, it just fits. It's the way she speaks, it's the way she thinks, it's the way she speculates.
My mother shouldn't read this book. She's pretty darn liberal-minded, but she's an elegant lady, and the language would be off-putting for her. I know that. So Mom, don't bother with this. Honestly. You won't like it.
And another thing, Mom: You're well aware that our senses of humour, at least in printed material, don't always align. Remember how Dad and I used to howl over Calvin and Hobbes when that comic strip was a regular feature in the Brandon Sun? You just didn't get why we found it funny. And you were frequently perplexed when Grant (my brother) and I used to die laughing when I read him Dave Barry stuff. So I know you won't like this.
But I think a good chunk of the population will.
That said, I move, finally, to the wine connection. Reading the book stone-cold sober, I still went into hysterics every single time. But reading it while sipping a glass of wine was a different experience. And that's not because the wine made Lawson any funnier. She doesn't need any help in the hilarity department.
The wine came into play because of my uncontrollable reaction to what I read. And really, you'd think I would have learned NOT to combine wine with "Let's Pretend." Because as I discovered, different wines elicit different sensations — when you pass them through your nose!!
That's right. I laughed so hard — more than once — that the near-unthinkable happened. When I laughed, it was usually with such force that I managed to snort at the same time, and wine rushed up to and through and out my nose. That's an experience I don't care to repeat. Ever.
Because while some of the wines hurt less than others, none of them traversing those tender nasal pathways was even remotely pleasant. However, some hurt less than others. But only slightly.
So if you're going to read "Let's Pretend This Never Happened — A Mostly True Memoir," and you insist, as I did, on sipping wine while you do so, I have a couple of suggestions.
First, the 2011 Wonderwall Sauvignon Blanc Semillon ($16.06) from Australia is a very pleasant beverage to enjoy while reading. It smells and tastes of tropical fruit, and while it's a bit crisp, the Semillon softens the often-highly-acidic Sauv Blanc substantially, making it a great compromise for someone like me who's not always taken with ultra-bright acidity.
And while it's not so great in the through-the-nose department, it was the least painful of any of the wines that travelled that same route.
Forget about reds. Those tannins were killers.
Another tip: The lower the alcohol content of the wine, the less it hurts when it goes where it's not supp ... oh. Yes. One other thing.
In addition to the wine/nose episodes, there were some unintentional, uncontrollable spit-takes. And I was reading "Let's Pretend It Never Happened" on the company iPad.
Understandably, it's not in the same shape it used to be.
Sorry about that, Curt. (He's my boss.) We need to talk.
» Diane Nelson is a long-time journalist and former Sun staffer who really likes wine. A lot. Chat with her online at www.vinelines.ca


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Distant Echo
Our mission at Distant Echo is a simple one; to sell quality designer clothing that broadcast a football casual and cultural message to discerning gents.
We understand there's a certain breed of man who are engrained in football and music culture. Us included. We're proud of what we wear and have a staple helping of clothes that celebrate this obsession of what we're about.
You'll enjoy free UK delivery on clothing, same day despatch on orders placed before 4.30pm and no quibbles returns.
We're proud to stock brands for the casual inclined, including the hugely successful and influential 80s Casuals and Casual Connoisseur as well as brands producing the highest quality wares at the top of their game such as Peaceful Hooligan, COPA and Weekend Offender.
@distantechouk on Instagram
Distant Echo Blog
Nov 30
Exclusive New T-Shirt from The Tea Street Band
We've never hid our support for The Tea Street Band over the years and we're proud today to receive stock of an exclusive new t-shirt from the lads. ... [Read more]
Nov 20
Black Saturday Liverpool Pop Up Shop
Exclusive offline discounts for one day only with our pop up store at Liverpool's Hard Times & Misery.Massive savings to be had on current and pre... [Read more]
Distant Echo New Arrivals
- Mundial Magazine - Issue 6 £7.00
Save 45%Available Sizes:The Northern Boys Club Riot - Navy £13.75 £25.0
Save 27%Available Sizes:Casual Connoisseur - The Casuals - White £22.00 £29.99


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Amazingly Effective Monitor Isolation Pad with Anti-recoil Design!
The Primacoustic RX9 Recoil Stabilizer flat monitor pad eliminates resonant coupling from the the nearfield speaker to the stand, giving you the most accurate sound possible from your monitor. The RX9 also provides a stable base for your monitor, with a non-slip neoprene top surface and multi-layer, multi-material construction. With the added mass of a laser-cut steel plate sandwiched by isolation foam, plus a high-density urethane foam base, the anti-recoil design of the RX9 is both stable and amazingly effective.Primacoustic RX9 Recoil Stabilizer Flat Nearfield Monitor Pad Features:
- Anti-recoil design
- Non-slip neoprene top surface
- High-density urethane foam base
- Laser-cut steel plate sandwiched by isolation foam for added mass
- 15" x 11"
- Flat
- Load limit 50 lbs


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