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Conserved rules govern genetic interaction degree across species
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DOI: 10.1186/gb-2012-13-7-r57
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- Koch, E.N., Costanzo, M., Bellay, J. et al. Genome Biol (2012) 13: R57. doi:10.1186/gb-2012-13-7-r57
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Synthetic genetic interactions have recently been mapped on a genome scale in the budding yeast Saccharomyces cerevisiae, providing a functional view of the central processes of eukaryotic life. Currently, comprehensive genetic interaction networks have not been determined for other species, and we therefore sought to model conserved aspects of genetic interaction networks in order to enable the transfer of knowledge between species.
Using a combination of physiological and evolutionary properties of genes, we built models that successfully predicted the genetic interaction degree of S. cerevisiae genes. Importantly, a model trained on S. cerevisiae gene features and degree also accurately predicted interaction degree in the fission yeast Schizosaccharomyces pombe, suggesting that many of the predictive relationships discovered in S. cerevisiae also hold in this evolutionarily distant yeast. In both species, high single mutant fitness defect, protein disorder, pleiotropy, protein-protein interaction network degree, and low expression variation were significantly predictive of genetic interaction degree. A comparison of the predicted genetic interaction degrees of S. pombe genes to the degrees of S. cerevisiae orthologs revealed functional rewiring of specific biological processes that distinguish these two species. Finally, predicted differences in genetic interaction degree were independently supported by differences in co-expression relationships of the two species.
Our findings show that there are common relationships between gene properties and genetic interaction network topology in two evolutionarily distant species. This conservation allows use of the extensively mapped S. cerevisiae genetic interaction network as an orthology-independent reference to guide the study of more complex species.
Gene Expression Omnibus
Gene Ontology
new end take off
Synthetic Genetic Array.
Most genes are not essential for eukaryotic life under standard laboratory conditions, which may reflect that organisms are highly buffered from genetic and environmental perturbations [1]. However, rare combinations of singly benign genetic variation can lead to synergistic effects, such as synthetic lethality, where mutations in two genes, neither of which is lethal independently, combine to generate an inviable double-mutant phenotype [2]. Because natural variations that distinguish two people occur relatively frequently [3] and complex genetic interactions may underlie most individual phenotypes [1], understanding the general principles that govern genetic networks may be critical for solving the genotype-to-phenotype problem and implementing personal medicine [4].
Recently, we tested approximately 5.4 million Saccharomyces cerevisiae gene pairs for genetic interactions, mapping an extensive network of more than 100,000 interactions by synthetic genetic array (SGA) analysis [5]. The study discovered both negative genetic interactions, instances in which a double mutant exhibits a more extreme phenotype than the expected combined effect of the single mutants, as well as positive genetic interactions, instances in which a double mutant exhibits a less-pronounced phenotype than expected [6]. This study revealed the distribution of genetic interactions with respect to gene function, highlighting a central role for chromatin-related, transcription, and secretory functions. Additionally, it identified several fundamental physiological and evolutionary gene properties that are significantly correlated with genetic interaction degree in the S. cerevisiae genetic interaction network [5]. For example, we showed that the genetic interaction degree of a gene is highly correlated with single mutant fitness, such that genes with a substantial fitness defect show a large number of genetic interactions.
While genetic interactions have been the most extensively studied in the yeast S. cerevisiae, there is intense interest in developing and applying large-scale screening technologies in other species. For example, large studies have already been completed in Escherichia coli [7, 8], Schizosaccharomyces pombe [9, 10], Caenorhabditis elegans [11, 12], Drosophila melanogaster [13, 14], and human cell lines [15, 16, 17]. Although definitive comparative analysis of these networks across species would be premature given the sparsity of known interactions in species other than S. cerevisiae, there have been preliminary comparative studies. In particular, the yeast S. pombe provides an attractive setting for this analysis due to the availability of a genome-wide deletion mutant collection [18] and scalable technology for automated genetic analysis [2]. Furthermore, S. cerevisiae and S. pombe are estimated to have diverged approximately 500 million years ago and display markedly different physiological properties [19] but share 75% of their gene content [19, 20]. The two comparative studies to date estimated approximately 30% conservation of individual negative genetic interactions, but also found substantial differences between the two species [21, 22]. These studies demonstrate the power and necessity of comparative analysis of genetic interaction networks, but have conducted only limited sampling of genetic interactions in S. pombe. The properties of these networks that are conserved across species and the rules governing their evolution remain largely open questions, making further characterization of the evolution of genetic interaction networks important.
In this study, we build predictive models capturing the relationship between gene properties and genetic interaction network connectivity, demonstrating that a small set of properties can accurately predict degree in the S. cerevisiae genetic interaction network. We show that models built from the genome-scale S. cerevisiae genetic interaction network also successfully predict genetic interaction degree of S. pombe genes, the vast majority of which have not previously been screened for interactions. Finally, we use our model to predict differences between the S. pombe and the S. cerevisiae networks, some of which may be reflective of functional rewiring and physiological differences between the species, and show that these differences are independently supported by the divergence of co-expression networks based on comparative analysis of gene expression.
Results and discussion
Modeling interaction degree in the S. cerevisiaegenetic interaction network
Pearson correlations between features and negative genetic interaction degree in S. pombe (pom) and S. cerevisiae (cer) are observed to be significant in many cases
Pearson's r
SM fitness defect
[0.41, 0.54]
[0.77, 0.74]
[0.22, 0.37]
[0.29, 0.23]
[-0.01, 0.15]
[0.19, 0.13]
Broad conservation
[-0.09, 0.08]
[0.19, 0.12]
PPI degree
[0.06, 0.33]
[0.19, 0.12]
Expression level
[-0.13, 0.03]
[0.14, 0.08]
[0.04, 0.21]
[0.14, 0.08]
Codon Adaptation Index
[-0.1, 0.06]
[0.12, 0.06]
Protein length
[-0.08, 0.09]
[0.08, 0.02]
Co-expression degree
[-0.08, 0.09]
[0.08, 0.02]
Number of domains
[-0.1, 0.07]
[0.05, -0.02]
Number of unique domains
[-0.1, 0.07]
[0.04, -0.03]
[-0.1, 0.07]
[-0.05, -0.11]
[-0.14, 0.04]
[-0.07, -0.14]
Copy number
[-0.17, 0]
[-0.08, -0.15]
Expression variation
[-0.23, -0.07]
[-0.15, -0.21]
To this end, we applied a regression tree approach to model combinations of 16 gene features that are predictive of negative genetic interaction degree (Figure 1b). Regression trees are built by repeatedly splitting sets of training genes, according to the values of gene features, until genes are sorted into small sets that each contain genes with similar genetic interaction degrees. The hierarchy of gene sets produced is visualized as a binary tree and the final sets of genes are each associated with linear regression models that assign predictions to query genes (Figure 1b). Bootstrapped subsets of the training data were used to build an ensemble of regression trees; this use of multiple models, bootstrap aggregation, is a typical method for building a robust predictive model [23] (Materials and methods).
To validate our approach, we used our model to predict negative genetic interaction degree for all genes in the S. cerevisiae genetic interaction network (Figure 1c; Materials and methods). A high correlation (r = 0.80, P < 10-324) was observed between predicted and actual genetic interaction degrees of genes not used in training the models, indicating that our model accurately reflects topological features of the S. cerevisiae genetic interaction network (Figure 1c). A strength of this type of model, in addition to providing degree predictions for previously unseen genes, is that the learned tree structures highlight rules consisting of combinations of gene features that explain variation in degree (Figure 1b).
Predicting genetic interaction degree in a distantly related species
Despite the low conservation of single mutant fitness and the varying correlations between individual gene properties for orthologs, we found that relationships between S. pombe gene features and genetic interaction degree were strikingly similar to those observed in S. cerevisiae (Figure 2b, Table 1). Consistent with S. cerevisiae trends (Figure 1a, Table 1), fitness defect was the strongest predictor of S. pombe genetic interaction degree. That is, S. pombe strains with severe fitness defects often exhibit high numbers of genetic interactions. The observed trends suggested that in addition to correlation with individual gene features, higher-level combinations of features that are predictive of connectivity in the S. cerevisiae genetic interaction network [5] (Figure 1a) may also be informative of S. pombe genetic interaction degree.
To test this hypothesis, we built a predictive model relating the combination of available gene features to genetic interaction degree in S. cerevisiae and then applied the resulting model to predict genetic interaction degree in S. pombe (Materials and methods). Interestingly, we observed significant correlation (r = 0.51, P < 10-36) between interaction degree predicted by our model and the number of interactions previously determined [10] for 548 S. pombe genes (Figure 2c, left side, light blue bar).
Our ability to predict genetic interaction degree from a small set of gene-specific properties is evidence that rules governing genetic interaction network topology are conserved across a large evolutionary distance (Figure 2c). Importantly, there is no significant decrease in correlation between predicted and actual interaction degree when predictions were restricted to genes unique to S. pombe (Figure 2c, left side, dark blue bar), indicating that the model performs equally well when applied to genes lacking orthologs in the species used to learn relationships in the model.
As a baseline comparison for our cross-species predictive model, we built a model from S. pombe gene features and genetic interaction degrees instead of from S. cerevisiae data. Within-species predictions for S. pombe interaction degrees are not significantly more accurate than predictions made by the cross-species model (Figure 2c, left side, compare red and light blue bars). We also note that although a simplistic predictor that maps the degree of a S. cerevisiae gene directly to its S. pombe ortholog provides reasonable performance (Pearson correlation approximately 0.4), this strategy is out-performed by our cross-species model and is limited to conserved genes. Strikingly, the models trained on S. pombe interactions and features were also able to predict interaction degree in the S. cerevisiae network with high accuracy (Figure 2c, right side, compare red and light blue bars). In general, interaction degree predictions for S. pombe genes were weaker than S. cerevisiae interaction degree predictions, which may reflect the limited functional diversity of available S. pombe genetic interaction studies [9, 10]. Nonetheless, the ability to predict interaction degree using features measured in either yeast species is evidence that relationships between genetic interactions and fundamental physiological and evolutionary properties are generally conserved.
The strong correlation between single mutant fitness defect and negative genetic interaction degree has the unsurprising consequence that the models are considerably influenced by this feature. To explore the reliance of our model on fitness defect, we constructed two types of bootstrapped regression tree models that were trained on all features except fitness defect. The first of these additional models is trained to predict negative genetic interaction degrees and is able to successfully make both within- and cross-species predictions (Figure S1 in Additional file 1). The second model was trained to predict the residual negative genetic interaction degree that remained after subtracting degree predictions made from a regression tree model that was trained on the single feature single mutant fitness defect. The prediction of these residuals by the remaining features was also significant (Figure S2 in Additional file 1). We therefore consider the inclusion of many other features to be a worthwhile part of our model, since they capture aspects of genetic interaction degree that fitness defect alone does not.
Validating predictions with S. pombewhole-genome screens
Consistent with our results for a published dataset [22] (Figure 2c), we observed a significant correlation (r = 0.40, P < 10-111) between the predicted number of interactions and the total number of experimentally derived interactions observed for a given array mutant in this genome-wide deletion set. Grouping genes with the same observed degree, we found that the distributions of our predictions were reflective of actual degrees (Figure 3b). For example, the median degree percentile predicted for genes with a degree of one was approximately 0.72, while the median prediction for genes with zero interactions was approximately 0.42. Importantly, the significance of the correlation was robust to the choice of interaction cutoff and persisted for a higher-confidence, sparser network (Materials and methods).
Identifying network rewiring suggested by cross-species predictions
Although many individual genes are conserved, yeast genetic interaction networks may have undergone substantial rewiring, as only approximately 30% of the interactions are conserved [9]. Similarly, a low conservation of genetic interactions has also been observed between S. cerevisiae and C. elegans [25]. To examine the extent of network rewiring, we first inferred interaction degrees for the entire S. pombe genome using our cross-species model. Because the predictions do not depend on sequence orthologs (Figure 2a, c), they can be used to compare the topologies of the S. cerevisiae and S. pombe networks even though only a small fraction of the S. pombe network has been screened.
However, we also identified many examples of possible rewiring, in which a significant difference in network connectivity, observed in S. cerevisiae and inferred in S. pombe, was found for orthologous modules (Figure 4a; Figure S3 in Additional file 1; Materials and methods). These predicted-rewired groups represent complexes or biological processes that may have evolutionarily diverged in terms of their importance in the genetic interaction network, acting as hubs in one species but not in the other. In particular, we found that 11 of 65 (17%) protein complexes and 44 of 545 (8%) GO biological processes may have undergone significant rewiring (Figure 4a; Figure S3 in Additional file 1) at a level of significance expected to identify only 3 and 27 (5%) rewired modules, respectively. For example, components of the dynactin complex are hub genes in the S. cerevisiae genetic interaction network (complex average of 85th percentile; Figure 4a) whereas the orthologous genes were predicted to exhibit average connectivity in the S. pombe genetic interaction network (complex average of approximately 50th percentile; Figure 4a). Dynactin, a multi-subunit protein complex known for interacting with dynein and enabling long-range movement along microtubules (reviewed in [27]), has been implicated in a S. cerevisiae cell cycle checkpoint pathway that arrests cell cycle progression in response to perturbations in cell wall synthesis [28]. A similar checkpoint has not been reported in S. pombe, suggesting that the difference in the number of genetic interactions observed across species may reflect a dynactin-specific role in monitoring S. cerevisiae cell wall integrity.
In addition to S. cerevisiae-specific genetic interaction hubs, we also identified gene groups predicted to be hubs in the S. pombe but not observed as such in the S. cerevisiae genetic network. One such case is the calcineurin-associated protein complex (Figure 4a). A difference in network connectivity might reflect a unique role for calcineurin in the regulation of bi-polar growth activation in S. pombe [29]. Unlike an S. cerevisiae cell, which grows predominantly via an actin-dependent budding mechanism, an S. pombe cell grows in a highly polarized bi-polar manner from its two ends. Following cell division, cell growth is initiated from the old end first, and later, after completion of S phase, from the newer end that forms at the site of cell septation (referred to as new end take off, or NETO). Calcineurin has been shown to play an important role in the delay of NETO by directly dephosphorylating critical targets involved in microtubule dynamics at the site of cell growth. This mechanism is dependent on activation of Cds1 kinase, best known for its role in the intra-S phase DNA replication checkpoint [30]. A connection between the intra-S phase checkpoint and inhibition of bipolar growth activation is so far unique to S. pombe and distinct from the checkpoint controls operating in S. cerevisiae. Additionally, calcineurin is dispensable for growth in S. cerevisiae [31]; in S. pombe, its deletion leads to defects in cell growth, cytokinesis, cell polarity, mating, and spindle pole body positioning, which are widespread effects consistent with its hub-like activity [32].
While our method of identifying rewired modules reports several statistically significant differences, we note two caveats in interpreting these results. First, since degrees of genes within functional modules may be systematically poorly predicted, our procedure may incorrectly identify modules as significantly rewired in cases where our test statistic would also have indicated that the within-species difference between predicted and observed degree was significant. Therefore, as a control, a version of this rewiring experiment that compares observed and predicted S. cerevisiae degrees will enable identification of cases that do not reflect true cross-species rewiring (Figure S4a, b in Additional file 1). Second, due to variations in the experimental protocol for measuring genetic interactions, there are differences in the media on which fitness defects were measured in S. cerevisiae and S. pombe, which may also contribute to apparent rewiring [33].
Functional properties of genes can be captured by many types of biological networks, so we turned to an independent dataset for confirmation of our rewiring predictions. To enable a comparative analysis of gene expression profiles across the two yeasts, we constructed a species-specific S. pombe co-expression network using a previously published approach [34] and large collections of publicly available expression data (Materials and methods), and obtained a previously published S. cerevisiae network [35]. Each species' network contains 832 genes that are one-to-one orthologs between the two yeasts and connected genes are those pairs that have high co-expression values surpassing a threshold of the 95th percentile. At our selected density of 0.05, there are approximately 17,000 edges in each network. In general, we found evidence of conservation between the S. cerevisiae and S. pombe networks: co-expression edges between two genes occurred in both networks for 9.2% of the gene pairs that were co-expressed in at least one network. This is about twice the background conservation rate of approximately 4.3%, as determined through comparison to a randomized network produced by a degree-preserving procedure.
To explore the connection between genes predicted to be rewired in the genetic interaction networks and differences between the co-expression networks, rewiring predictions were overlaid on the co-expression networks. Specifically, all non-essential one-to-one orthologs were classified as either rewired or non-rewired based on our prediction of genetic interaction degree (Figure 4b). Using this rewiring labeling, we measured the conservation rate of three types of co-expression edges: co-expression edges connecting two non-rewired genes, connecting two rewired genes, and connecting rewired and non-rewired genes.
We found that co-expression edges involving predicted rewired genes are consistently less conserved than edges with exclusively non-rewired endpoints (Figure 4c), a trend that is robust over different co-expression thresholds used for network sparsification (Figure S5 in Additional file 1). For example, when genes whose degrees differ by 55 interactions or more are considered rewired, 6.9% of the co-expression relationships connecting rewired genes are conserved (107 of 1,659), in contrast to the significantly higher 10.1% of co-expression relationships that are conserved between non-rewired genes (1,238 of 12,472, Fisher's exact test P < 10-6). This trend grows stronger when considering genes that were predicted to have even larger differences between S. pombe and S. cerevisiae. This analysis independently confirms predictions of highly rewired genes between the two species and suggests that changes at the level of gene expression regulation are at least one mechanistic factor that contributes to these differences.
Although individual interactions and gene-specific properties may not be strongly conserved between species, our findings suggest that these properties influence genetic interaction networks in a similar manner. For example, while the genes important for normal growth may vary, the relationship between a gene's fitness contribution and the genetic interactions it exhibits appears to be conserved. Indeed, models trained on both S.cerevisiae- and S.pombe-derived gene properties were significantly predictive of cross-species genetic interaction degree (Figure 2c), suggesting that the general principles governing genetic interaction network structure are retained through evolution. Thus, a complete genetic interaction network for an organism such as S. cerevisiae should serve as a reference network to guide studies to uncover genetic interactions in more complex systems. Predicting specific pairwise interactions across species is of course the next (more difficult) challenge, but models that can accurately predict the variation in number of interactions across the genome provide a foundation for cross-species interaction analysis. Our results also demonstrate that integrative comparisons leveraging multiple functional genomic datasets across species may be one approach to build confidence in differential network analysis. As more data become available, both the extent and nature of network conservation should reveal how functional conservation and divergence can be recognized and utilized in distantly related species.
Materials and methods
Gene features
Yeast conservation
Yeast conservation is a count of how many of 23 different species of Ascomycota fungi possess an ortholog of a given gene. This measure was first described in [36], and ortholog data were downloaded from [37]. The 23 species are an expanded set of the 17 species described in the study, with the additions of Schizosaccharomyces octosporus, Schizosaccharomyces japonicus, Lodderomyces elongosporus, Candida parapsilosis, Candida tropicalis, and Candida guilliermondii.
Broad conservation
Similar, though complementary, to yeast conservation, broad conservation is a count of how many out of a set of 86 non-yeast species possess an ortholog of a given gene. To count this, we obtained orthogroup designations from InParanoid [38]. For each gene, we considered it to have an ortholog in another species only if it appeared in a cluster with the other species and was given a score of 1.0 by the InParanoid clustering method; that is, we considered a yeast gene to have an ortholog in species x if it was a seed gene for a gene cluster that had an orthologous cluster in species x. Although Ostlund et al. [38] considered 100 species, we disregarded the yeast species, since the yeast conservation measure already captures information from these species.
Codon Adaptation Index
The Codon Adaptation Index, a measure of bias in the usage of synonymous codons, was calculated with the cai tool in the EMBOSS suite [39]. For each gene, the index is based on a comparison between codon frequencies in the gene and frequencies observed in a set of highly expressed genes; for both S. pombe and S. cerevisiae, EMBOSS included a default codon usage table that was used.
Copy number
Copy number is a count of the number of paralogs a gene has. This was determined from clusters identified by the InParanoid algorithm [40] run on S. cerevisiae and S. pombe. All genes that appear in the same cluster were considered copies.
The protein disorder measure is the percent of unstructured residues in a gene's protein product as predicted by the Disopred2 software [41].
dN/dS is the ratio between nonsynonymous and synonymous mutations in coding regions of genes. For S. pombe genes, dN/dS was calculated twice, using S. japonicus, S. octosporus as out-group species, and averaged to produce a final dN/dS estimate. Orthologous protein sequences were globally aligned with EMBOSS [39] using default parameters. For each S. pombe gene, only the out-group ortholog that produced the highest alignment score was used for dN/dS calculations; dN/dS ratios were calculated with the PAML package's implementation of the Yang and Nielsen method for estimating substitution rates [42, 43].
Similarly, we computed the average dN/dS ratio for S. cerevisiae in comparison to the sensu strictu yeast species (Saccharomyces paradoxus, Saccharomyces bayanus and Saccharomyces mikatae). Protein sequences were aligned using MUSCLE [44] and dN/dS ratios were computed using PAML [42].
Number of domains
The number of domains for a gene is the number of regions that Pfam has identified as domains in the protein sequence of the gene. Domain matches for each protein were obtained online from the Pfam database [45].
Number of unique domains
Since the same domain is often repeated multiple times in a single protein, this feature modifies number of domains by counting the number of unique domains present in each protein.
This measure is a simple statistic of codon usage bias and expresses the effective number of codons used in a gene. The chips tool of EMBOSS [39] was used to calculate this feature.
Protein length
Protein length is simply the number of amino acids in the corresponding protein.
Co-expression degree
This measure is derived from the co-expression network, the construction of which is described in its own section. The network contains a level of co-expression for all pairs of genes. We therefore sparsified the network by considering only edges between gene pairs whose co-expression levels were above the 95th percentile. The co-expression degree of a gene is the number of genes with which its co-expression value is retained in this restricted network.
Expression level
Expression variation
We estimated the amount of variability in a gene's expression level by measuring the variance of its expression across a number of different microarray experiments, which included microarray data from different growth conditions and replicates. Within each study, we found each gene's percentile of variation. The final value assigned to each gene is its average percentile across all studies. These datasets were obtained from a number of different studies that deposited data in the Gene Expression Omnibus (GEO) [49]. S. pombe data used in this analysis are the same as those used in construction of the S. pombe co-expression network.
Fitness defect
S. pombe fitness defect measurements were obtained by conducting a series of control SGA experiments as described elsewhere [2, 9, 33]. Briefly, a S. pombe SGA query strain harboring a dominant drug-resistance marker (natMX4) inserted at a neutral genomic locus (h- leu1Δ::natMX4 ade6-M210 ura4-Δ18 leu1-32) was crossed against the S. pombe non-essential deletion mutant collection (h+ geneXΔ::kanMX4 ade6-M210 ura4-Δ18 leu1-32). Following mating and sporulation, haploid meiotic progeny harboring both the kanMX4 and natMX4 markers are selected and colony sizes are measured after applying standard normalization procedures. We have previously shown that colony sizes derived from these control screens reflect fitness defect of the kanMX4-marked single mutant strains that comprise the deletion mutant array. Fitness estimates were based on four control screens as described above and combined with five mutant screens (prz1, res2, SPAC1687.22c, SPCC1682.08, and SPAC6G9.14), which contained the dominant drug-resistance marker (natMX4) [9].
S. cerevisiae fitness defect values, defined quantitatively in [6], were published in [5] and experimental procedures are detailed in [33]. As in the S. pombe protocol described above, SGA was used to insert a neutral query marker into mutant strains so that we could observe colony growth for each mutant in the deletion collection under the effects of only the single deletion. Fitness estimates are based on a large number of replicate screens.
Protein-protein interaction degree
The protein-protein interaction degree of each gene's protein is the number of physical interactions reported in BioGRID, version 2.0.58 [50]. Interactions considered physical were restricted to those identified by the following terms: Affinity Capture-MS, Affinity Capture-RNA, Affinity Capture-Western, Biochemical Activity, Co-crystal Structure, Co-fractionation, Co-localization, Co-purification, Far Western, FRET, PCA, Protein-peptide, Protein-RNA, Reconstituted Complex, and Two-hybrid.
Multifunctionality is a measure of the number of GO terms that are annotated to a gene [26]. From GeneDB [51] and Saccharomyces Genome Database [52] gene association files (download in November 2009) for S. pombe and S. cerevisiae, respectively, redundant terms - one term from pairs of terms that are considered 'alternative ids' - were removed before totaling the number of GO term annotations for each gene.
Genetic interaction degrees
Negative genetic interaction degrees of S. pombe genes were derived from interactions reported in [10] (Additional file 2). Only those interactions with S-scores ≤ -2.5 were considered. This dataset contains 551 genes that are involved in chromosome function; intentionally included are approximately 100 genes that participate in processes present in both S. pombe and human, but importantly, are not present in S. cerevisiae (for example, RNA interference machinery).
Negative genetic interaction degrees of S. cerevisiae genes (Additional file 3) were collected from the measurements reported in [5], which screened for interactions involving 3,456 array genes, 1,438 of which have S. pombe orthologs. As suggested by the authors, only negative interactions with an epsilon value of ≤ -0.08 and a P-value cutoff < 0.05 were considered. This dataset includes degree measurements for most non-essential genes.
Orthology mappings (Additional files 4 and 5) are from the InParanoid eukaryotic ortholog database [24]. Although the InParanoid algorithm produces clusters, our analysis depends on ortholog pairs. To calculate correlations between S. cerevisiae and S. pombe for each of the gene features (Figure 2a), only genes in one-to-one orthology mappings were used. When holding out orthologs for degree prediction in a set of 'species-specific' genes (Figure 2c), all genes that had any number of orthologs were removed. Since InParanoid may not report orthologs that other algorithms have detected, we took a conservative approach by additionally removing any genes that had an ortholog in the pombe database GeneDB [53], which includes manually curated orthologs.
Models and evaluation
Our models are bagged regression trees that use the 16 features described above (Additional file 6). Breiman [23] suggests that using an ensemble of only 25 classifiers can result in nearly all improvement gains that bagging can produce over a single classifier; however, we used 100 trees because the computation required in training is relatively low and we were interested in analyzing the tree structures. Individual trees were trained by MATLAB's classregtree function, which minimizes node impurity according to mean squared error. For each tree, a bootstrap sample was used to select, with replacement, a set of training genes the same size as the set of total genes (therefore each tree is trained on approximately 63.2% of all genes) and held out genes. The final prediction for a single gene of the species used to train the model (that is, the within-species prediction) is the median of all predictions from trees for which the gene was not in the training set (Additional file 7). The final prediction for a gene of the species not used to train the model (that is, the cross-species prediction) is the median of predictions from all trees (Additional file 8).
To assess the performance of the model, we calculated the Pearson correlation coefficient between predicted and actual degrees of genes with known degrees. To estimate stability of performance, we repeated the model construction and evaluation 25 times and reported predictive ability as the mean Pearson correlation coefficient and its standard deviation across all 25 repetitions for within- and cross-species cases (Figure 2c).
S. pombegenetic interaction screens
Eight whole-genome S. pombe genetic interaction screens were completed using the method described in [9]. The query strains were deletion mutants for each of the following genes: SPCC1682.08c, SPBC21D10.12, SPBC13E7.09, SPAC4G8.13c, SPAC3A11.13, SPAC27D7.13c, SPAC22F3.09c, SPAC16A10.07c. The resulting double mutant colonies were processed as described in [6]. Negative interactions were derived from the scores by applying an interaction cutoff of ≤ -0.08 and a P-value cutoff of < 0.05. Degree measurements were then derived for all non-essential genes by counting the number of significant interactions across the set of eight queries (Additional file 9). Significant correlation with the predicted degrees was also observed when a stricter cutoff was applied (interaction score ≤ -0.12, P-value < 0.05 yielded a correlation r = 0.41, P-value < 10-117).
Rewiring groups and significance assessment
To make comparisons between degrees of orthologs in the genetic interaction networks of the two yeast species, we considered genetic interaction degree to be predicted percentile for all S. pombe genes, while percentiles of actual degrees were used for S. cerevisiae.
To search for groups of functionally related genes that have been rewired since the divergence of S. pombe and S. cerevisiae, we defined gene groups in two ways. The first simply grouped genes whose protein products form a complex in a set of complexes defined in [6] (Additional file 10). The number of proteins per complex ranges from 2 to 81, with the vast majority having 6 or fewer proteins.
The second method for making sets of functionally related genes grouped genes that share a biological process GO term annotation [26] (Additional file 11). We considered GO terms that are annotated to greater than 3 and fewer than 50 genes in either of the two species. Additionally, a group of S. cerevisiae genes was required to have a minimum number of two genes with known genetic interaction degrees; a group of S. pombe genes was required to have a minimum of two genes with known fitness defect. Since GO terms tend to be highly redundant, we filtered gene groups so that no pair of groups overlapped by more than 50% of either group's genes.
To determine orthologous pairs of groups that have significantly different average degrees, we calculated the difference between the median degrees of genes in each species' group, and then compared the differences to a distribution of differences produced from randomly grouped genes. We generated this background by creating groups of randomly selected genes in one species, then identifying orthologous groups in the other species composed of the selected genes' orthologs. A query gene-group pair was compared to a background containing only random gene-group pairs whose group sizes were identical to the query groups. For example, a protein complex of five individual S. cerevisiae proteins may contain four genes that have S. pombe orthologs; this query gene-group pair would be compared with a background of groups with five random S. cerevisiae genes matched with a group of four of their S. pombe orthologs.
Comparative analysis of co-expression networks
To independently validate genetic interaction degree differences across species, we performed a comparative analysis of co-expression networks of S. cerevisiae and S. pombe genes. The S. cerevisiae network was previously published [34] and is based on integration of a large collection of expression datasets. To construct the S. pombe network (Additional file 12), data from nine expression studies were collected from the GEO database [54] (Additional file 13). Genes with missing values for more than 30% of the samples were removed, and the remaining missing values in each dataset were imputed using KNNImpute [55]. Datasets reflecting probe intensities (rather than relative ratios) were log-transformed. After processing, the nine S. pombe expression datasets were integrated as described in [34, 56]. The naive Bayes approach for dataset integration requires a gold standard set of positives, for which we used direct gene co-annotation to any term in the GO that contained between 2 and 100 genes. S. pombe gene annotations were downloaded from the GO website [26, 57] in May 2011. All analysis and integration of expression data were completed using the Sleipnir library [56].
We applied a 95th percentile cutoff to edges in both the S. cerevisiae and S. pombe co-expression networks, such that only the highest scoring 5% of edges were retained.
To estimate the overlap between the S. cerevisiae and S. pombe networks in the absence of biological conservation, we randomized the edges of the S. cerevisiae network and considered the background conservation to be the overlap between this randomized network and the S. pombe network. The randomizing procedure repeatedly chose two random edges and exchanged an endpoint of one edge with an endpoint of the other edge, thus maintaining the degrees of genes in the network. The number of endpoint swaps performed was 20 times the number of edges in the network, which is a sufficient number of swaps to remove the original relationships between genes.
EK is an NSF Graduate Research Fellow. EK, JB, RD and CLM are partially supported by a grant from the Minnesota Partnership for Biotechnology and Medical Genomics program, a grant from the National Institutes of Health (1R01HG005084-01A1), and the National Science Foundation (DBI0953881). EK, MC, JB, RD, CLM, BJA, and CB are also supported by a grant from the National Institutes of Health (1R01HG005853-01). MC, BJA, and CB are supported by the Canadian Institutes of Health Research (MOP-57830) and the Ontario Research Fund (GL2-01-22). GC was supported by funds from a CIHR-Operating grant. KC-R was supported by a Queen Elizabeth II Graduate Scholarship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors thank Jeff Piotrowski for providing S. pombe query strains, Karen Dowell and Matt Hibbs for providing co-expression network software, Valerie Wood for providing orthology maps, and Tahin Syed for help with image processing.
Supplementary material
Copyright information
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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PRESS DIGEST - Wall Street Journal - March 21
March 21 (Reuters) - The following are the top stories in the Wall Street Journal. Reuters has not verified these stories and does not vouch for their accuracy.
* U.S. President Barack Obama ordered a second round of financial sanctions on Russian officials and a St. Petersburg bank over Crimea, and the European Union added names to its sanctions list. Moscow responded with penalties against nine U.S. officials. (link.reuters.com/zes77v)
* The Federal Reserve's annual test of big banks' financial health showed the largest U.S. firms are strong enough to withstand a severe economic downturn, a sign that many will get the green light soon to reward investors by raising dividends and buying back shares. The Fed said 29 of the 30 largest institutions have enough capital to continue lending even when faced with a hypothetical jolt to the U.S. economy lasting into 2015, including a severe drop in housing prices and a spike in unemployment. (link.reuters.com/bus77v)
* Symantec Corp, the computer-security company synonymous with "antivirus," fired Chief Executive Steve Bennett amid sagging revenue and slumping shares, the second time it has dumped a CEO in less than two years. (link.reuters.com/dus77v)
* Airbnb Inc, which once sold novelty cereal boxes to stay afloat before emerging as a threat to the hotel industry, is close to becoming one of the world's most valuable startups. The online service that lets people rent their homes to travelers is in advanced talks to raise capital that would value it at about $10 billion. (link.reuters.com/hus77v) Continued...

subset: commoncrawl


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Delivery: A mix of daytime, evening, Saturday, and online courses.
Start: Fall or Spring Semester, Summer Session, Full- or Part-Time.
- Liberal Arts and Sciences — AA Degree (60 credits)
- Minnesota Transfer Curriculum — MnTC (40 credits)
- Liberal Arts Pathway
About the Discipline
Learning a foreign language gives you a window into cross-cultural understanding by providing you with a fundamental tool: the ability to communicate directly with speakers of other languages. In addition to teaching you communication skills, foreign language classes deal with current and historical cultural, socio-political, and economic aspects of the regions where the language is spoken. Studying Spanish at Century College:
- gives you a greater awareness and deeper understanding of other cultures.
- prepares you for travel for business, education, or pleasure.
- broadens your career opportunities.
- helps you appreciate the aesthetics of literature, music, art, and folklore of other cultures.
- helps you develop more flexibility in thinking processes through problem solving, conceptualizing, reasoning, and dealing with ambiguity.
- enables you to transfer four semester courses (Beginning and Intermediate Spanish) to a four-year university.
- connects you to the Spanish-speaking community through service learning projects.
Points of Pride
Century College's Language Laboratory offers students the opportunity to practice the Spanish language in a comfortable setting. Language lab resources include computers to access audio programs and the Internet, videos, dictionaries, readers, maps, and current magazines.
Spanish Club activities include viewing and discussing films, fundraising events, weekly salsa dance instruction, museum and restaurant visits, celebrations of special days, and the opportunity to meet and hear speakers from Spanish-speaking countries.
In addition, Spanish language students are introduced to the Resource Center of the Americas (located in Minneapolis.) The Center's goal is "to promote human rights, democratic participation, economic justice, and cross-cultural understanding in the context of globalization in the Americas."
Related Links
Transfer Opportunities
Thinking of transferring to a Bachelor's degree program in Spanish? Century offers high-quality, affordable options--from individual courses to 2-year degrees.
Start your Spanish bachelor's degree at Century. Many students complete the Associate in Arts degree (AA) and develop an individual academic plan for their intended major, such as Spanish, including all the pre-major requirements and lower-division major requirements. The AA degree can generally be considered the first two years of a four-year degree and transfers as all your lower-division general education requirements completed to all Minnesota public colleges and universities, as well as many private and out-of-state colleges and universities.
Please see your Advisor or Counselor for assistance in developing your individual academic plan and transfer pathway for your educational goals.


text:
Sons of Thunder (Part One)Mature
In 1941, four extremely different teen boys arrive at a British boarding school. Herschel is a Polish Jew and ghetto escapee, haunted by his past. Erich is German and an ex-Nazi Youth. Jim is an American traveling England in search of adventure. And Gabe is a British-born Italian, a violinist and a closeted homosexual. They have all landed at Wellington's School for Boys, where they are forced together as roommates. The story follows the trials of their unlikely friendship through the Holocaust.
Herschel's Log
Sunday, March 9, 1941
I'm sitting on a bus full of strangers in a country I first saw a week ago. I know that this is the wrong place to begin my story, but if I don't calm down I'll probably throw myself out the window (which I imagine would only make things worse) and at this point writing is the only way for me to calm myself down.
Rebeccagave me this journal back in London. She shouldn't have spent the money, but I'm glad I have it now. I miss my typewriter back home, but this will have to work for now. I hope Rebecca is all right alone, but I guess she's handling the sudden change of scene a lot better than me anyway. The whole time we were in London I couldn't help but feel like I needed her more than she needed me. I need someone to get me through this, and without her I don't know what I'll do.
I sure as hell won't be able to turn to anyone here. I'll never tell anyone about what happened back home. It's only not that I don't want anyone to know, but I just can't imagine talking about it. It will kill me if I ever have to say it out loud. I can't even write it down. No one can know about Kristen.
I can see the gates of Wellington's now. I've been waiting to see this place for the past two hours, for my whole life, really. Dad's talked about it for as long as I can remember. But I don't relish it all. It's not that I miss where I came from. There is nothing on earth that could make me go back. But though the past few weeks have been excruciating, they've gone incredibly fast. I don't know if I can handle the world I've been tossed into. I need to make myself out to be a fairly normal student in about five minutes, and all this time alone with my thoughts isn't exactly keeping me sane.
But Poland is far behind me, at least literally, and I know I have to leave everything else behind too.
The bus is stopped at the entrance. Apparently I'm the only one getting off here, so I guess they're waiting for me. I don't know what I'll find behind the gates, but, like it or not, I'm going to find out.


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Courtyard by Marriott Atlanta Marietta Windy Hill
Select your stay dates to check availability at Courtyard by Marriott Atlanta Marietta Windy Hill
Courtyard by Marriott Atlanta Marietta Windy Hill
About the Hotel
Property Location
Located in Marietta (Cobb Galleria), Courtyard by Marriott Atlanta Marietta Windy Hill is convenient to Cobb Galleria Centre and Cumberland Mall. This hotel is within close proximity of Cobb Energy Performing Arts Centre and Fox Creek Golf Course.
Make yourself at home in one of the 127 air-conditioned rooms featuring flat-screen televisions. Complimentary wireless Internet access keeps you connected, and cable programming is available for your entertainment. Partially open bathrooms with shower/tub combinations feature complimentary toiletries and hair dryers. Conveniences include phones, as well as safes and desks.
Take advantage of recreational opportunities offered, including a spa tub, a 24-hour fitness center, and a seasonal outdoor pool. Additional features at this hotel include complimentary wireless Internet access and a fireplace in the lobby. Guests can catch a ride to nearby destinations on the complimentary area shuttle.
You can enjoy a meal at a restaurant serving the guests of Courtyard by Marriott Atlanta Marietta Windy Hill, or find a snack in a coffee shop/café. Quench your thirst with your favorite drink at a bar/lounge. Cooked-to-order breakfasts are available daily for a fee.
Business, Other Amenities
Featured amenities include a business center, express check-in, and express check-out. Event facilities at this hotel consist of conference space and meeting rooms. Free self parking is available onsite.
Hotel Facilities
- Conference space
- Accessible bathroom
- Breakfast available (surcharge)
- Terrace
- Free newspapers in lobby
- Snack bar/deli
- Restaurant
- Meeting rooms 2
- Number of floors - 3
- Total number of rooms - 127
- Free self parking
- Bar/lounge
- Safe-deposit box at front desk
- Spa tub
- Luggage storage
- Smoke-free property
- 24-hour front desk
- Dry cleaning/laundry service
- Free area shuttle
- Elevator/lift
- Airport transportation
- Coffee shop or café
- Free WiFi
- Laundry facilities
- Roll-in shower
- Coffee/tea in common areas
- In-room accessibility
- Express check-in
- Express check-out
- Outdoor seasonal pool
- Fireplace in lobby
- 24-hour fitness facilities
- Business center
Room Facilities
- Daily housekeeping
- Free cribs/infant beds
- Iron/ironing board
- Desk
- In-room climate control (air conditioning)
- Phone
- Partially open bathroom
- Free WiFi
- Cable TV service
- Coffee/tea maker
- Free toiletries
- Air conditioning
- In-room safe
- Refrigerator (on request)
- Shower/tub combination
- Hair dryer
- Pay movies
- Flat-panel TV
- Premium TV channels


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A splendid example of the third case was laid before the Sixth European Congress on Biotechnology held in Florence last month. They heard of a bacterium, isolated from a mollusc related to clams, scallops and oysters, which produces an enzyme that may well find an important role in biological washing powders.
Like the enzymes already incorporated in some detergents, it attacks proteinaceous stains - such as blood, egg, grass and sweat - that adhere strongly to textile fibres. But it can also break down proteins over an unusually wide range of temperatures, and is specially suited for washing powders because it lacks the sodium tripolyphosphate now being eliminated on environmental grounds.
The concept of enzyme detergents goes back to the beginning of this century, when chemists first incorporated protein-digesting enzymes (proteases), extracted from pancreatic glands, in laundry pre-soaks. They hoped that trypsin and other proteases would break down protein stains, but these experiments were only partially successful.
The first commercially successful biological detergent became possible in the early Sixties when the Danish company Novo launched a product known as Alcalase. The protease in this was not only effective in digesting protein stains. It was also unaffected by the other components of washing powder, and worked at then routine high temperatures. The Dutch firm Kortman & Schulte, in collaboration with Gebruder Schnyder in Switzerland, used Alcalase in Biotex, a breakthrough in enzyme detergents. A temporary setback in the Seventies, when the enzyme powders caused allergic reactions in a few factory workers, was soon overcome by encapsulating the enzymes in a tough coating.
The discovery announced in Florence arose from studies at the Agricultural Research Service in Peoria, Illinois. Richard Greene and two colleagues at the US Department of Agriculture, Harold Griffin and Michael Cotta, were investigating the digestive system of the marine shipworm Psiloteredo healdi - not a true worm, but an elongated mollusc that can bore into woodwork, often of ships, by rotating its two shells. The broken-down wood and anything else encountered passes into the shipworm, where it is digested in the Gland of Deshayes (named after the French geologist Gerard Paul Deshayes, who in the 19th century first observed the gland in fossil molluscs).
It was no surprise some years ago when other researchers found that the Gland of Deshayes contained enzymes capable of breaking down the cellulose fibres in wood, yielding glucose which the shipworm uses as a source of energy. The enzymes are formed not by the mollusc itself, but from bacteria living in the gland; just as bacteria inside ruminants, such as cows and sheep, form enzymes that digest the cellulose in grass and other plants.
But the shipworm does not live on cellulose alone. It requires enzymes to break down other materials, including proteins, ingested during its boring activities. Mr Greene and his co-workers discovered that these include a protease secreted into the gland by an as yet unnamed bacterium that lives there. This protease is active and stable at temperatures between 25C and 50C, and is equally effective in attacking proteinaceous stains when used as a pre-soak or for the wash itself.
One of the most attractive properties of the newly discovered enzyme stems from its high activity in the sort of alkaline conditions found in the Gland of Deshayes.
For many years, sodium tripolyphosphate has been used in washing powders to prevent various forms of dirt from adhering to fabrics. But as it has come under increasing suspicion as a cause of the deadening of lakes and rivers by eutrophication, this chemical has been removed from commercial laundry detergents, or at least drastically reduced in concentration.
The new formulations tend to be relatively alkaline, a condition in which existing proteases are less effective. But the activity of the new shipworm proteases is not impaired in this way. The enzyme is also expected to work well in the energy-conserving cold-water detergents that are being developed.
The Peoria researchers even have a second application for their new enzyme, which they have shown can be produced relatively cheaply by growing the shipworm bacterium in nutrient broth. It is highly effective in removing the protein that accumulates on contact lenses. And, unlike existing proteases used for this purpose, it remains active in the presence of the disinfectant hydrogen peroxide. A single solution can thus be used in place of the conventional two solutions for cleansing and sterilising.
Can you imagine any funding agency in Britain these days backing a scientist who wanted to help contact-lens wearers by studying the digestive system of a so-called worm that isn't even a worm anyway?
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Rich and historic past connected to the national museum of the motorcar and to its founders, the Schlumpf family, a prestigious hotel was built during the roaring twenties and the entirely decorated in its original style. The hotel Du PARC is in Alsace the reference of the luxury hostelry where cro...
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Terri Proud's Issue Positions (Political Courage Test)
What is the Political Courage Test?
Arizona State Legislative Election 2010 Political Courage Test
|a) Do you consider yourself pro-choice or pro-life?
|b) Should abortion be legal only within the first trimester of pregnancy?
|c) Should abortion be legal when the pregnancy resulted from incest or rape?
|d) Should abortion be legal when the life of the woman is endangered?
|e) Do you support requiring parental notification before an abortion is performed on a minor?
|f) Do you support requiring parental consent before an abortion is performed on a minor?
|g) Do you support sexual education programs that include information on abstinence, contraceptives, and HIV/STD prevention methods?
|h) Do you support abstinence-only sexual education programs?
|i) Other or expanded principles
I believe parents have the responsibility to teach their children about sex related issues, not the schools. Schools should focus on educating our children with the basics of education. As for the unanswered abortion issues, I believe women need to be educated more on the decision and have ALL the options.
1) State Spending
Indicate what state funding levels (#1-6) you support for the following general categories. Select one level per category; you may use a number more than once.
|Slightly Increase
|a) Education (higher)
|Slightly Increase
|b) Education (K-12)
|c) Environment
|Maintain Status
|d) Health care
|Greatly Increase
|e) Law enforcement
|Slightly Decrease
|f) Transportation and highway infrastructure
|Slightly Decrease
|g) Welfare
|h) Other or expanded categories
In Arizona we need to look at why schools are receiving over 50% of the state's budget including the taxes from direct sources that do not go through the Legislation and not able to push even .40 cents into the classrooms or pay our teachers. The problem isn't funding, the problem is that it stops at the administration level.
2) State Taxes
Indicate what state tax levels (#1-6) you support for the following general categories. Select one level per category; you may use a number more than once.
|Slightly Increase
|a) Alcohol taxes
|Maintain Status
|b) Cigarette taxes
|Slightly Decrease
|c) Corporate taxes
|Slightly Decrease
|d) Gasoline taxes
|Slightly Decrease
|e) Property taxes
|Slightly Decrease
|f) Sales taxes
|g) Income taxes (low-income families)
|h) Income taxes (middle-income families)
|i) Income taxes (high-income families)
|j) Other or expanded categories
In the state of Arizona we need to implement a flat tax and reduce our Corporate taxes. We should not be looking for ways to raise our taxes in a time when families are losing jobs, and losing their homes. We need to focus on bringing more tax payers in Arizona by bringing business to our State.
3) Budget Stabilization
Indicate which proposals you support (if any) for balancing Arizona's budget.
|a) Issuing the early release of certain non-violent offenders
|b) Increasing tuition rates at public universities
|c) Instituting mandatory furloughs and layoffs for state employees
|d) Reducing benefits for Medicaid recipients
|e) Privatizing certain government services
|f) Other or expanded principles
End wasteful spending. For example: We should end the tax revenue sharing. There is nothing in our Constitution that government should be collecting taxes on behalf of the cities and towns. Giving this responsibility back to the cities and towns would save the state $550 million.
|a) Do you support limits on the number of terms for Arizona governors?
|b) Do you support limits on the number of terms for Arizona state legislators?
c) Do you support limits on the following types of contributions to candidates for state government?
|1) Individual
|2) Political Action Committee
|3) Corporate
|4) Political Party
|d) Should candidates for state office be encouraged to meet voluntary spending limits?
|e) Do you support requiring full and timely disclosure of campaign finance information?
|f) Other or expanded principles
Arizona has Term Limits for governors and State Legislators now. On question (c) I believe any candidate that runs for office should be able to receive donations from any tax payer and taxpaying entity.
|a) Do you support capital punishment for certain crimes?
|b) Do you support alternatives to incarceration for certain non-violent offenders, such as mandatory counseling or substance abuse treatment?
|c) Should the possession of small amounts of marijuana be decriminalized?
|d) Should a minor accused of a violent crime be prosecuted as an adult?
|e) Should a minor who sends sexually-explicit or nude photos by cell phone face criminal charges?
|f) Do you support a comprehensive ban on text messaging while driving?
|g) Do you support the enforcement of federal immigration laws by state and local police?
|h) Other or expanded principles
|a) Do you support reducing government regulations on the private sector?
|b) Do you support the reauthorization of the payday lending industry?
|c) Do you support increased state funding for job-training programs that re-train displaced workers?
|d) Do you support expanding access to unemployment benefits?
|e) Do you support providing financial incentives to the private sector for the purpose of job creation?
|f) Do you support increased spending on infrastructure projects for the purpose of job creation?
|g) Do you support providing direct financial assistance to homeowners facing foreclosure?
|h) Other or expanded principles
I personally believe that the pay lending industry should be illegal in the state of Arizona. I believe that if a tax payer loses a job, faces foreclosure then the state should offer assistance by reforming welfare to lending help to taxpayers who are in need of it.
|a) Do you support national education standards?
|b) Do you support requiring public schools to administer high school exit exams?
|c) Do you support using a merit pay system for teachers?
|d) Do you support state funding for charter schools?
|e) Do you support the state government providing college students with financial aid?
|f) Should illegal immigrants who graduate from Arizona high schools be eligible for in-state tuition at public universities?
|g) Other or expanded principles
F. I need to way the pro's and con's to this prior to giving an answer
|a) Do you support state funding for the development of alternative energy?
|b) Do you support state funding for the development of traditional domestic energy sources (e.g. coal, natural gas, oil)?
|c) Do you support providing financial incentives to farms that produce biofuel crops?
|d) Do you support state funding for improvements to Arizona's energy infrastructure?
|e) Do you support state funding for open space preservation?
|f) Do you support enacting environmental regulations aimed at reducing the effects of climate change?
|g) Other or expanded principles
|a) Do you support restrictions on the purchase and possession of guns?
|b) Do you support requiring background checks on gun sales between private citizens at gun shows?
|c) Do you support allowing individuals to carry concealed guns?
|d) Do you support requiring a license for gun possession?
|e) Other or expanded principles
B. I believe many vendors at the gun shows already participate in background checks.
|a) Do you support a universally-accessible, publicly-administered health insurance option?
|b) Do you support expanding access to health care through commercial health insurance reform?
|c) Do you support interstate health insurance compacts?
|d) Should individuals be required to purchase health care insurance?
|e) Do you support monetary limits on damages that can be collected in malpractice lawsuits?
|f) Do you support legalizing physician-assisted suicide in Arizona?
|g) Do you support allowing doctors to prescribe marijuana to their patients for medicinal purposes?
|h) Other or expanded principles
B and G would need more research.
|a) Should marriage only be between one man and one woman?
|b) Should Arizona allow same-sex couples to form civil unions?
|c) Do you support state funding for stem cell research?
|d) Do you support state funding for embryonic stem cell research?
|e) Do you support the state's use of affirmative action?
|f) Do you support the inclusion of sexual orientation in Arizona's anti-discrimination laws?
|g) Do you support the inclusion of gender identity in Arizona's anti-discrimination laws?
|h) Other or expanded principles
I believe individuals should be employed based on their level of performance rather than race, sex, etc. That to me is discrimination.
|Please explain in a total of 100 words or less, your top two or three priorities if elected. If they require additional funding for implementation, please explain how you would obtain this funding.
1. Jobs. We HAVE to focus on bringing jobs into this state if we are going to have a future. In order to do that we MUST create a business friendly environment.


text:
ED Magedson – Founder
Ripoff Report
BFC PRODUCE INC.P.O.Box 740 Lenoir, North Carolina USA
BFC PRODUCE INC. Bernhardt company I took a servey for mistery shopper and recieved a check to do shopping and that check was returned to my bank cousing an over drafted Internet
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Fix it the right way.
Corporate Advocacy Program™
i filled out a survey for mistrey shopper on line to shp in stores like wal-mart, home depot, k-mart. they sent me a check for 1970.00 dollars i put it in my bank. the bank said that check was sent back to them. now i am over drifted that amount.
This report was posted on Ripoff Report on 08/01/2013 11:02 AM and is a permanent record located here: http://www.ripoffreport.com/reports/bfc-produce-inc/lenoir-north-carolina/bfc-produce-inc-bernhardt-company-i-took-a-servey-for-mistery-shopper-and-recieved-a-che-1072099. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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Weingut Franz Schindler
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- Breakfast Available
- Air-Conditioning
- Free parking
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TripAdvisor Reviews Weingut Franz Schindler Mörbisch
Travel Blogs from Mörbisch
1st Day in Bratislava
... told me a bit about the city and told me how to reach to the tourist information center. There they gave me a map, even though not very necessary as the old town is quite small and it's better to get lost around.
So it was about 17:00 and I left the apartment. First thing I saw was the blue church, as it was very close. A nice church painted in blue, originally related to the school next to it. There was a mise, so I could ...
Cycling round Vienna
... for €12. Lucy was taken aback when she was charged €62, but was glad to hear €50 of that was a deposit and would therefore be returned. Having bikes has been really great. We've seen more of the city than we would have by simply walking and even had the occasional race to be second in line. Katie was our 'cycle director', as she named herself, so she automatically took Alpha position. Massive shout out to our leader for leading ...
Venice here we come!
... Continued into Italy and as before not too far into a new country you see the changing style of houses industries etc. After leaving bus taken to hotel on a water taxi. A real character hotel small Old furniture very cute but good bed and air conditioning! Very hot-35deg' A quick gondola ride complete with music serenading ... Sensational ...
Vienna Day 2
... of music (it was filmed in Salzburg for goodness sake). Much to the boys relief we moved away from the Sisi museum to visit the Spanish horse stables. These are famous horses that are trained in dressage and reside in the castle. After the stables we arrived at an enormous building from which a very famous photograph of Hitler addressing the Austrian people was taken (it is taken from behind Hitler and looks out at the massive crowd he is addressing). It is ...
... might imagine. It took us about 20 minutes to find the center. We saw this maze on the Amazing Race this past season, which made it even cooler to run through. After finding our way out, we trekked up the hill and viewed Vienna from above (see the pictures). Vienna is gorgeous! Most of the people we have come across are friendly and, again, the language barrier hasn't been a problem (yay)! We take off for Budapest tomorrow via a 2.5 hour train ride and plan to spend about 3 days ...


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BJ'S Brewery and Restaurant - Fresno, CA
in Brewpubs
BJ's Brewery is a small brew pub at the Fashion Fair Mall in Fresno, CA.
posted by: saopaulo1
location: California
date approved: 1/6/2011
last visited: never
Patronsky pivovar - Bratislava, SVK
"Patrón svetlý 11,5°" non-pasteurized beer from minibrewery
Patronsky pivovar.
"Patrón svetlý 11,5°" nepasterizovane pivko z Patronskeho minipivovaru.
posted by: walden2
location: Bratislavský kraj, Slovakia
date approved: 1/4/2011
last visited: 7/31/2013
San Tan Brewing Co - Chandler Arizona
A fine brewpub in historic downtown Chandler Arizona
posted by: sundevil1994
location: Arizona
date approved: 1/3/2011
last visited: 9/23/2015
Blackbeck Brewery, Egremont, Cumbria
Blackbeck Brewery is a family run small brewery in a purpose built simple design in the pub car park. Behind the Blackbeck Inn on A595.
posted by: flipflopnick
location: Northwest England, United Kingdom
Husumer Brauhaus, Husum
Theodor-Storm-Hotel und Husumer Brauhaus.
posted by: Grizzly-NF
location: Schleswig-Holstein, Germany
last visited: 1/25/2017
Pivovarský dvur Purkmistr, PM, CZ, EU
Brewpubs in brewery Purkmistr in Pilsen Cernice.
posted by: Arogant
location: Plzensky kraj, Czech Republic
last visited: 3/29/2014
Heartland Brewery, New York City, New York
HEARTLAND BREWERY AND ROTISSERIE: New York City's most famous landmark, the Empire State Building, now houses New York's number one brewery, a two-story 450-seat Heartland equipped with two large bars and four giant flat-screen TV's.
posted by: SUp3rFM & Cruella
location: New York
date approved: 12/7/2010
last visited: 11/10/2016
The Strands Inn, Nether Wasdale, Cumbria
The Strands Inn has brewed its own beer since 2007, producing two ales sold on the premises. Real fires and real ale, a winning combination.
date approved: 11/30/2010
last visited: 4/19/2013
Brewpub Joyce -Caldas da Rainha – Portugal
Brewpub Joyce -Caldas da Rainha – Portugal Nossa Senhora do Pópulo R Engenheiro Cancela Abreu 11, Caldas Rª 2500-197 CALDAS DA RAINHA Telefone: 262 824 294
posted by: h_raky
location: Leiria, Portugal
date approved: 11/20/2010
last visited: 1/5/2014
Dogfish Brewpub - Rehoboth Beach, Delaware
Dogfish Head Brewings & Eats in downtown Rehoboth Beach, Delaware....
posted by: muffy53
location: Delaware
date approved: 11/8/2010
last visited: 7/5/2014
TOMO BRÄU - RT-Betzingen, Germany
Kleines Restaurant mit Brauerei / Small restaurant with brewery
posted by: Family Fog
location: Baden-Württemberg, Germany
date approved: 10/16/2010
last visited: 4/22/2017
Cannery Row Brewing Company - Monterey, California
The Cannery Row Brewing Company is located in the Cannery Row area of Monterey, California.
posted by: GT.US
last visited: 11/8/2014
Lone Peak Brewery - Big Sky, MT
Located in the "Brewski" building at the base of Big Sky Ski Resort, Lone Peak Brewery offers 6 regular year-round beers, and 5 seasonal beers.
posted by: CerealBoxMonsters
location: Montana
Pepperwood Bistro, Brewery & Catering
A great brewpub in Burlington, Ontario.
posted by: MrPirateCat
location: Ontario, Canada
date approved: 9/27/2010
last visited: 11/27/2013
Goldfinch Brewery - Tom Browns - High East Street, Dorchester, Dorset, UK
Tom Brown's public house brew their own beer in the Goldfinch Brewery behind the pub.
posted by: Dragontree
location: South West England, United Kingdom
Granite City Brewpub
Welcome to Granite City!
posted by: Lord Elwood
location: Indiana
date approved: 9/19/2010
last visited: 8/30/2012
Ram Restaurant and Brewery - Salem, Oregon
Popular watering hole close to Oregon's capitol.
posted by: shadow101
location: Oregon
date approved: 8/31/2010
last visited: 4/6/2013
Blackfoot River Brewing Company - Helena, MT
Brewers of IPA, OPA, Stout, Steamboat Lager, Amber Ale, Wolly Bugger, Cream Ale, Scottish Ale, and Bitter.
date approved: 8/11/2010
last visited: 6/15/2013
Cock & Bull English Pub & Brewery - Lynfield, Auckland, New Zealand
A Traditional English Pub & Brewery providing award winning ale and delicious food plus entertainment.
posted by: No Fixed Aboder
location: North Island, New Zealand
date approved: 8/9/2010
Zámecký pivovar Chyše
A minibrewery with pub and restaurant at Chyse chateau.
posted by: haggaeus
location: Karlovarsky kraj, Czech Republic
date approved: 7/9/2010
last visited: 11/9/2015
Elliot Glacier Public House - Parkdale, OR
The Elliot Glacier Public House lies in the shadow of the beautiful Mt. Hood in Parkdale, Oregon, USA.
posted by: Marine Biologist
date approved: 7/3/2010
last visited: 2/20/2012
Brandy Cask Pub
Brandy Cask and Brewary
posted by: Deadjesters
location: West Midlands, United Kingdom
date approved: 6/16/2010
last visited: 8/9/2015
Piolho D'Ouro
posted by: K!nder
location: Porto, Portugal
date approved: 6/6/2010
last visited: 3/5/2011
Golden Valley Brewery and Pub - McMinnville, Oregon
A popular brewpub in downtown McMinnville.
date approved: 6/4/2010
last visited: 6/2/2010
Greenodd Brewery at The Ship Inn, Greenodd, Cumbria
Greenodd Brewery is the Ship Inn's own brewery producing two barrels each brew.
date approved: 5/31/2010
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Refresh your look for spring with bold lippies, fun nail art and major brows.
The Best "It" Braid For Your Hair Length!
Sorry, winter, we're way over you. It's time to refresh your look for spring with bold lippies, fun nail art and major brows—here's how to rock the hottest new trends, just like these stylish celebs!More
Big, full brows are still having a major moment! This season, it's more about emphasizing your natural shape than overly filling them in, so just use a little brow powder to amp up sparse spots, then brush through.
Try: Eyebrow Kit, $10.80, forever21.com
MORE: 8 Celebs With Amazing Eyebrows
Stark white nails feel totally fresh, just like the crisp spring weather! Try a matte or textured finish to rock two of our favorite polish trends at once.
Try: Deborah Lippmann Nail Lacquer "Pseudo Silk Kimono," $20, sephora.com
Vanessa Hudgens's Amazing Beauty Makeover!


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Basketball Wallpaper
by: Minions • 0
All Android applications categories
by: Minions • 0
Basketball Wallpaper
A good start and a dunk shot in a ball ! Best basketball wallpaper ! Your favorite sport equip your cell phone screen. Choose your favorite basketball wallpapers and feel to replace it when you want to use the new. Take your time !
- Easy-to -navigate interface that allows you to find the required !
- Custom wallpaper that suits your taste and screen !
- UN zoom, zoom, rotate, move , zoom, adjust brightness, add various effects , and save your wallpaper design !
Basketball is a popular sport . This game is the most common in the United States . Each team has five players, each team trying to get a basketball, on the other side of the net. You play this game a ball, which is a sphere orange with black stripes. Have a professional team , and university teams. High school basketball is very common , and even some teams in some primary form . The main thing is to get the ball into the opposing team's hoop , with your hands dribbling and shooting .
There are several different ways that you can score points. The first is a large basket extends past the foul line . In this range any shooting is worth 2 points. Anything other than a large arc shot is worth three points . If you're shooting foul, free throw line you can go to earn your point of view . Each throws a minute. There are many rules, including some of the rules are added to each new year . It is played with indoor and outdoor tennis courts.
Courts and the size of the basket height will vary . The length of time the game is played also varies according to age and skill level of players.
This is a fast moving game involving many species, including shooting and dribbling . Adoption or defense involves a lot of fast movement.
It involves a lot of starts and stops . Although not renowned as an aerobic exercise , it is still a good exercise that can help you burn calories ( the game one hour can burn 630-750 calories ) .
The great thing about the game is that you need very little additional equipment to play . However, because it involves a lot of running and jumping , it can be wise to have basketball or sports shoes, provides good ankle support. Protective knee and elbow pads are very useful.
Play this game is a great way for kids active and have fun. " Basketball Australia " is a primary school child, started by Basketball Australia and the Australian Sports Commission National referral program . It provides elementary school girls and boys ages , abilities and skill levels to play in a safe environment and develop social skills. Download wallpapers and enjoy your favorite sport !
Tags: basketball wallpaper
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Arich dance legacy was assured in Cuba many years ago by the three mythical figures who founded the Cuban National Ballet: Fernando Alonso, his younger brother Alberto, and Fernando's wife, Alicia Alonso.
Fernando Alonso was born in Havana on 17 December 1914 and died there, at the Cardiovascular Hospital, on 27 July. Fernando and Alberto were among the few and first men to train as classical ballet dancers under the guidance of their mother, Laura Raineri. The news of his death was broken to us during the final performance of A Tribute to Rudolf Nureyev by the distinguished Cuban ballerina and teacher Loipa Araujo – associate artistic director, with Tamara Rojo, of the English National Ballet.
Fernando Alonso Raineri, the elder of two sons of Matías Alonso and Laura Raineri, was brought up in a house filled, like his country, with music and dance. His mother was a pianist and later president of Havana's Pro Arte Musical, which brought the young boy into contact with some of Cuba's – and the world's – greatest musicians. He also loved science and nature.
It was Alberto who first started ballet classes, to help make him a better footballer. Soon Fernando joined him, and stayed – especially since there were 126 girls to six boys, and one of those young girls was Alicia Martínez del Hoyo. The two soon fell in love. In 1937, Fernando, seven years older than Alicia, went to the US looking for work, and Alicia, only 16 at the time, joined him. They got married and a year later their only daughter, Laura, was born.
In New York the couple danced continuously, first in musicals, then with some of the biggest names in the international dance world of that era, all the while taking classes with famous teachers. Fernando would study the different teaching practices of the different teachers, and took the best parts of each system to develop them, from a scientific foundation, into the basis of his own Cuban-flavoured pedagogical studies.
The couple danced all over the world together and were members of Ballet Theatre (as the present American Ballet Theatre used to be known then) for eight years. They never intended to stay in New York and returned home permanently in 1948 when Ballet Theatre found itself facing a financial crisis.
The desire to form their own company was financially challenging. When the money ran out, they advertised in a newspaper for a sponsor and La Polar brewery agreed to pay for one performance, providing there was some advertising on stage! "We told them we'd put bears on the stage, beer bottles, whatever they wanted," reported a member of the University Student Federation of Havana, which had pledged to support the company.
The Ballet Alicia Alonso persevered and, filled with dancers from Ballet Theatre, won acclaim on its tours of Central and South America. It was at this point that Fernando, while still performing, decided to change direction and dedicate himself to the needs of the company by becoming its director. However, dark days lay ahead for them all. During the Batista government, which Fernando, Alberto and Alicia opposed, the company was disbanded for three and a half years and not reinstated until the revolution of 1959.
In 1950, Fernando had claimed another milestone – that of founding the Alicia Alonso Academy of Ballet with the understanding that here, native Cubans could be trained in the art of classical ballet, and be a permanent source for the company. So it was on this project that all attention was focused during the years when the company did not function. With Fernando as teacher-director, all three Alonsos worked tirelessly in establishing an academy, which would – and still does – rival any of the top quality international ballet schools of today.
With the support of the Cuban Revolution, and under the aegis of Fidel Castro, Alicia, Fernando and Alberto Alonso created the foundations of the Cuban classical ballet company we now know as the Ballet Nacional de Cuba.
Fernando, the father of Cuban ballet, trained, among many others, the "Four Jewels" of Cuban ballet: Josefina Méndez, Mirta Pla, Aurora Bosch and Loipa Araujo – the latter considered one of the best ballet teachers in the world today. Famous dancers such as Rudolf Nureyev and Mikhail Baryshnikov sought to benefit from Fernando's classes whenever they could.
In 1975, after he and Alicia divorced, Fernando married the Cuban ballerina Aida Villoch and became the director of the Ballet School and Ballet Company of Camagüey, where he taught until very recently. He was awarded Cuba's National Dance Prize for Lifetime Achievement in 2000 and in 2008 he won the Prix Benois de la Danse, given to him at the Bolshoi Theatre in Moscow.
In her excellent biography, Fernando Alonso: The Father of Cuban Ballet, Toba Singer gives a full account of the life of an extraordinary, well-loved Cuban ballet figure.
He is survived by his daughter, Laura, his grandson, Iván, three great granddaughters, and Yolanda, his third and last wife.
Fernando Alonso, dancer and dance teacher: born Havana, Cuba 17 December 1914; married three times (one daughter); died Havana 27 July 2013.
- More about:
- Dance
- Fernando Alonso
- Music
- Performing Arts
- Pianists
- Theatre


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Park Basics
When available, place an Information Kiosk near the entrance of the park. Information Kiosks will sell maps, which will help keep some guests from getting lost. Don't forget that you can also sell umbrellas at those Kiosks; the guests don't have The Weather Channel everywhere they go.
You can now start building amusement rides. Build a good mix of rides, some gentle, some exciting, and of course a roller coaster. Place one, and only one, rollercoaster by the entrance to get the guests' hearts pounding early. It also increases their happiness. The rest of your roller coasters you will build deeper into your park (you build only one by the entrance since they will have to travel to the others, and will sometimes see other things on the way). This is how real amusement parks design their parks in case you didn't notice. You want at least one indoor type ride, like the Haunted House.
Now that you have some rides going, hire one of each type of employee. Uncheck the handyman's option to mow the grass. You don't need that yet. (mown grass looks nicer, and helps guests happiness ratings, and park value ratings) When you do want someone to mow the grass, hire a handyman specifically for that task, and uncheck all but the mow grass option.
You can now raise the entrance price to the park to around $15-$20. You want it cheap enough to keep your guests happy, but high enough to get you some money. Generally, for every 4 new rides that you build, add $5 to admission. $10 if one of those rides is a roller coaster. I'm not saying that the only money you get should be from the entrance fee, I still think you should also charge them on rides. This way you can maximize your chances of getting all your guests money.
Finally you are going to want to build some amenities, such as Bathrooms, and shops. Build these a little further away from the entrance, since the guests who enter don't need to use them yet. Avoid placing food shops near the exits of vomit intensive rides--although you can, and possibly should, place Drinks stands there.
Happiness - Happy people stay in your park and will be more willing to part with their money. Happiness is increased by fun rides, like roller coasters, and maintained by normal rides. The guests become unhappy if they are: Tired, Sick, Hungry, Thirsty, or have to go to the bathroom. Other things that contribute to unhappiness are: Long waits in line, and inability to find good rides, and how clean/pleasant your park is in appearance. Finally, don't forget that you do have Entertainers, and although it seems like they are worthless, they do help your guests happiness ratings.
Energy - A tired guest won't go on anything but the most exciting rides to "wake up", but once he has done that will go on other rides. Tired guests also walk slower than others, and are less happy. Place a good roller coaster by the entrance to wake up these sorry fellas.
Hunger, Thirst - These two stats start at a random value when the guest enters the park, usually under 1/4 full. These values increase mostly over time. Thirst is also affected by what the guest is doing. Eating Fries increases thirst, as does a high nausea rating.
Nausea, Nausea Tolerance - Guests always underestimate their chances of getting sick off of a ride. Nothing you can do about it except provide numerous benches, and clean any vomit up. Also note that sick guests become very unhappy and walk very slowly (to the point of slowing down the launching of rides).</dir>
As you go along, you will likely get bombarded with complaints from your guests. The guests want a nice clean park to visit, good rides to go on, and short lines. Also pricing things affordably gives the guest a good feeling that translates into happiness. When the guest likes something, they will give you the "good thought" (check the people button, and look under thoughts). Happy patrons, when they leave, promote your park to their friends.
Every once in awhile you will want to check the People button and look at a summary of their thoughts (the second "Group" tab). Checking here will give you a good idea of areas to improve in your park. If a lot of people complain of hunger, build some food shops; if people complain of having to go to the bathroom, build a bathroom; if lots of people complain about waiting in line forever, consider cutting the line down.
The best way to prevent Vandalism is to keep your guests happy. However, that doesn't always work, so here's some suggestions on vandalism. Whenever a guest goes red in the face, he is about to commit vandalism. So what do you do with a guest that you catch before he does anything? Put him in "Jail". That is, a one square path with a bench on it for him to smash. Let him smash it and get him outta there. Consider it "Destructive Therapy". If you are wondering how Security Guards affect vandals, Guests do not commit vandalism if there is a guard present. They don't actually get "caught".
Mechanics - As your rides age, their reliability will go down tremendously. To help alleviate this problem, swap the Inspection rate on the rides down to "every 10 minutes". At each inspection the reliability will go up a little. Also if your rides are REALLY old (5+ years) you need to take drastic measures to insure that they don't crash. Consider replacing the ride. Or, just reduce the number of trains on a rollercoaster down to 1. (this will avoid a crash) Another way to tell if it is time to replace a ride is "Down Time".
Handymen - For every roller coaster or shop that you build you should hire a new Handyman to help with the added trash and vomit. Make sure to "specialize" your handymen, that is, don't have them clean trash AND mow lawns, in that that is very inefficient. Have most your handymen clean trash, water gardens, and empty trash cans. Then have a few mow the lawns.
Entertainers - Entertainers are just that, entertainers. You put them in a costume of your choice, and they wander the park, and try to entertain the peeps in your park. One useful place for them is in the Queue. Some rides either have very long lines or the track is very long and needs more time to make it's way around. Entertainers can be used to cheer the waiting guests up.
Security Guards - You need security guards to stop Vandalism. But since you can't always stop it, you need to know what to do afterwords. Vandals strike only 3 things, benches, garbage cans, and lights. The vandalized bench will be broken in half, the garbage cans will be tipped over, and lamps will have their lights broken. Simply replace the old items with new ones to fix. (Don't right click to remove! That may get rid of the path as well) So who are all these vandals? If you check the people screen, the vandals are the ones with the red snarling faces. If you manage to catch one of these perps, you can try to get his anger out by placing him on a one square path with a bench. He'll smash the bench and become happy again (well... he won't be so violent anyway), then you can move him back into the park.</dir>
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R.I.P. David Brenner ..
David Brenner was one of the first comedians to whom I remember laughing out loud as a kid/teen. Jerry Seinfeld grew to much bigger fame, but Brenner set the standard Seinfeld copied. Rarely a curse word, always funny. He was also one of Johnny Carson's favorites .. appearing on the Tonight Show well over 100 times. This is one of them, from 1983 ..


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Anyone who operates a GMC knows that when it comes to maintaining their automobile only the most superior replacement parts will do. GMC car, truck, or SUV owners have become used to a certain level of reliability when driving their Yukon XL 2500 around town. When your GMC Yukon XL 2500 left the factory, it was destined for a driver who prioritizes trusted quality in their automobile. It's not all the time that you realize that you have to buy a replacement Brake Master Cylinder to repair your car or truck.
Using hydraulic pressure to accomplish a task is a millennia old feat of engineering. Your GMC Yukon XL 2500 has a brake master cylinder that creates a large amount of pressure for your braking system inside a very small space. This part uses the force from your brake pedal to compress brake fluid and distribute it to each caliper or wheel cylinder. The calipers and wheel cylinders are a form of slave cylinder; using the hydraulic pressure from the GMC Yukon XL 2500 brake master cylinder to pinch your brake pads against the rotor or the brake shoes against the drum so your vehicle can slow down if needed.
When your GMC needs a top quality replacement Brake Master Cylinder, there's no need to waste time looking anywhere other than the authority, Car Parts Discount. Given all the trouble that can be involved with performing maintenance on your car or truck, let www.carpartsdiscount.com be the number one store you go to make purchasing it easy. Make all your future car parts purchases at carpartsdiscount.com and realize incredible savings on all your upcoming repairs. When it comes to GMC Yukon XL 2500 Brake Master Cylinder parts, you can find them for 2013, 2012, 2011, 2010, 2009, 2008, 2007, 2006, 2005, 2004, 2003, 2002, 2001, 2000 models by clicking the appropriate year below.


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Click here to view sale items.
Can't find the item you are looking for using the search and browse tools above? View our special order catalog.
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- Inspired by old Italian Cafe Racers and clubman bars, adds interest to any bike
- CrMo-plated Tange steel
- Clamp diamater: 25.4mm
- Rise: 5 deg
- Width: 22.8" (579mm)
- Up/back sweep: 15 deg
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View other listings in Berkeley Vale, Bateau Bay, Long Jetty, The Entrance, Killarney Vale.
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I'm a young hard working female I am a apprentice boiler maker I stick to myself I don't smoke pretty quiet person easy going, currently looking to rent a room long term.
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Need Allergy Testing in Ozark, AL? Simply order online 24/7 and get fast, affordable Allergy Testing at a lab near you (Find More Locations):
Why would I get Allergy Testing?
Allergy Testing measures antibodies related to allergy in the blood. People in Ozark, AL order Allergy Testing for general screening or to guide therapy.
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Our Allergy Testing prices include the doctor's order that you need to get tested in Ozark, AL, all lab fees and taxes, and a pdf copy of your results. For current prices, please CLICK HERE.
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Kate Bosworth in Vogue
Designers Play Dress Up With Kate Bosworth in Vogue
Photo 1 of 15
Designers Play Dress Up With Kate Bosworth in Vogue
From simple and fresh to done up and sophisticated, Kate Boworth's Vogue spread is varied, but always gorgeous. Instead of a standard photo spread, Kate was styled by seven top designers including Proenza Schouler, Phillip Lim 3.1, Derek Lam, Rag & Bone and more. Lucky girl, right? She may be an actress, but she's certainly got that model quality. Which look is your favorite?
To see more of Kate in Vogue just


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Aviva Life - I-Life
About The Plan
In today's fast-paced life, rarely do we stop and take stock of the life cover we provide for our families in case something unfortunate was to happen to us. To ensure your families are secure and lead a comfortable life, in case you are not around, we bring you Aviva i-Life - a pure life insurance cover for your loved ones at a nominal cost.
To offer you convenience at your fingertips, you can buy this term plan online to save the time and effort that would have been otherwise required.
Key Features
Min. Sum Assured Rs. 25 lacs, Max. No Limit (subject to underwriting).
Entry Age: Min. 18 years, Max. 55 years.
Maximum maturity age 70 years.
Premium payment frequency-yearly and half-yearly.
Option to choose policy term from 10 to 35 years.
An additional rebate for female lives of 5% on the tabular premium rates for male lives.
Tax benefit u/s 80C & 10(10D) of the Income Tax Act, 1961.
Eligibility Criteria
- Minimum sum assured is 25 lakhs & maximum sum assured is No Limit (subject to underwriting)
- Minimum entry age is 18 years & maximum entry age is 55 years
- Minimum policy term is 10 years & maximum policy term is 35 years
- Premium payment mode is Half-yearly, Yearly
- Maturity benefits are Nil
- Grace period is 30 days
- Yes, under Section 80C & 10(10D) of the Income Tax Act, 1961
Note:- Taxes as applicable from time to time are payable on the premium, minimum premium is Rs. 25 Lacs Minumum age is 18 years and maximum age is 55 years.