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|Table of Contents
|Vol. 123, No. 1-4, 2008
Issue release date: March 2009
|Section title: Copy Number Variations in the Genome of Model Organisms
Copy number variation in the mouse genome: implications for the mouse as a model organism for human diseaseCutler G. · Kassner P.D.
Lead Discovery, Amgen, South San Francisco, CA (USA) Corresponding Author
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Individuals within a species have genetic differences which ultimately result in the spectrum of phenotypic variation that we observe. Genetic variation exists at the nucleotide level in the form of single nucleotide polymorphisms (SNPs), and at a structural level as inversions, deletions and amplifications of larger stretches of nucleotides. Profiling of human and mouse genomes has identified numerous genomic segmental copy number variations (CNVs) throughout these genomes. Since inbred mice are widely used laboratory models for the study of both normal and disease biology, it is crucial that we understand the full scope of genetic variation, including CNVs, within these animals. These genetic differences can inform us about the history of a population or species, enlighten us on gene function, and guide our selection of a model system for the study of human disease.
© 2009 S. Karger AG, Basel
The use of mice as research animals dates back to at least the 1800s. Their utility increased greatly at the turn of the 20th century with the development of the first characterized mouse strains by William Castle. This was soon followed by the creation of the first inbred mouse strain by Castle's student Clarence Little in 1909 (Paigen, 2003; Eisen, 2005). Contemporaneously, one of the first uses of mice to study human disease was the crossing of 'fancy mice' with 'common' laboratory mice to understand the heritability of resistance to transplanted tumor growth by E.E. Tyzzer (Paigen, 2003). Tyzzer's experiments were greatly complicated, however, by the genetic heterogeneity of his subjects. Using his new inbred mouse strain, Little solved this problem and conclusively demonstrated the Mendelian inheritance of transplanted tumor resistance loci.
This first strain, named DBA for the presence of the d (dilute), b (brown), and a (agouti) coat color alleles, was produced in the same manner as other inbred mouse strains that would come after, through repeated brother-sister crosses (and sometimes offspring-parent crosses) for at least 20 generations (Beck et al., 2000). This laboratory manipulation created a unique organism, a diploid mammal which simulated monoploidy by being homozygous at every (or nearly every) allele in the genome. These two characteristics, the phenotypic complexity of a mammal and a relatively static homozygous genetic background, made inbred mouse strains uniquely powerful models in which genetic studies of complex phenotypes could be carried out. In the years since the DBA strain was first generated, over 450 other mouse lines have been bred, although many of the most commonly used strains trace their history back almost as far as DBA (Beck et al., 2000).
Researchers have found proxies for various human diseases among this large number of disparate mouse strains, each with its own unique assortment of alleles and the phenotypes to which they give rise. While first used for studying cancer and immunology, the use of inbred mouse strains has become common in most, if not all, disease fields including the fields of study of developmental disorders, metabolic diseases, and neurologic and behavioral disorders (Bedell et al., 1997). In metabolic disease research, for example, strains have been identified with varying degrees of resistance or sensitivity to spontaneous and diet-induced diabetes, obesity, and atherosclerosis (Clee and Attie, 2007). These mice have long been used for Quantitative Trait Locus (QTL) mapping through the crossing of susceptible and resistant strains followed by serial back-crossing to identify the responsible genetic loci. More recently, rather than having to perform laborious back-crosses, the in silico technique of Whole Genome Analysis (WGA) has been developed (Grupe et al., 2001; Smith et al., 2003; Liao et al., 2004; Pletcher et al., 2004; Wang et al., 2005). In WGA, known sequence polymorphisms in existing inbred mouse strains are the foundation for the analysis. This technique opens the door to a much more time-efficient way to identify gene variants that affect phenotype, but it depends on prior characterization of a very large number of sequence polymorphisms. Fortunately, considerable work has already gone into cataloging large numbers of single-nucleotide polymorphisms (SNPs; Sherry et al., 1999) and this data served as the foundation for the studies referenced.
Despite the focus on SNPs for genetic mapping, they are not the only type of sequence polymorphism known. Although less common than SNPs, there exist amplifications and deletions of segments of the genome ranging from 1 kb to many megabases, known as segmental copy number variations (CNVs). CNVs may play an important role in both the range of normal mammalian phenotypic differences (Freeman et al., 2006) as well as in the etiology of disease (Gonzalez et al., 2005; Aitman et al., 2006). The potentially dramatic effects of large scale CNVs on human health have long been understood based on the identification of gross karyotype alterations (Emanuel and Shaikh, 2001; Shaw and Lupski, 2004), but it is only recently that a fuller picture of CNV content in mammalian genomes has started to emerge. With the advent of modern genomic technology, particularly array-based Comparative Genomic Hybridization (aCGH), it has become possible to do large scale surveys of the CNV content of genomes (Pinkel et al., 1998; Barrett et al., 2004). A number of researchers have done this work in humans (Sebat et al., 2004; Sharp et al., 2005; Redon et al., 2006; Zogopoulos et al., 2007; Jakobsson et al., 2008), identifying many hundreds of CNVs across diverse sets of individuals. Likewise, aCGH analysis has been brought to bear on the murine genome, exploring the CNV content of various sets of inbred mouse strains (Li et al., 2004; Snijders et al., 2005; Cutler et al., 2007; Egan et al., 2007; Graubert et al., 2007; She et al., 2008; Watkins-Chow and Pavan, 2008).
The earlier work on identifying mouse CNVs performed by Li et al. (2004; hereafter referred to as Li) and Snijders et al. (2005; hereafter Snijders) used BAC-based CGH arrays containing 19,200 BAC probes and 2,069 BAC probes, respectively. Li identified 347 BACs that probed genomic regions with apparent copy-number variations in 14 inbred mouse strains, using C57BL/6 as the reference strain. Snijders identified 79 BACs that probed apparent CNVs in seven inbred and one outbred mouse strain, using FVB as the reference strain. Unfortunately, due to the use of BAC-based arrays with low to medium genomic resolution, from 0.16 Mb per probe to 1.5 Mb per probe for these two studies, the actual borders of the CNVs were ill-defined as was the basic fact of whether or not adjacent altered probes represented one contiguous CNV. More worrisome, however, was the observation that comparing the results of these two studies revealed 'no obvious overlap' between the CNV-defining BAC probes (Snijders et al., 2005).
The most recent large-scale mouse CNV surveys moved away from using BAC-based aCGH to oligo-based aCGH (Cutler et al., 2007; Graubert et al., 2007; She et al., 2008; hereafter referred to as Cutler, Graubert, and She, respectively). Graubert compared 20 mouse strains to C57BL/6 using high-density NimbleGen oligonucleotide arrays. These arrays had over 388,000 probes with a density of more than one probe per 8 kb. Cutler, which was work done by our group, used high-density Agilent oligonucleotide arrays with approximately 244,000 probes to survey 41 strains of inbred mouse compared to C57BL/6 – the most comprehensive analysis done to date. She used a sequencing method along with an aCGH analysis of 15 strains on NimbleGen arrays. There was broad agreement between the Graubert and Cutler oligo-aCGH-based studies, with 67% of 72 CNV loci identified by Graubert also identified by Cutler (Cutler et al., 2007). However, neither of these two studies identified much, if any, overlap with the two BAC-aCGH-based analysis. For example, the Graubert CNVs matched only 7% (17 out of 238) and 4% (3 out of 74) of the CNV loci from Li and Snijders, respectively (Graubert et al., 2007). The lack of concordance between either of the BAC-based analyses and any of the other studies probably derives from a combination of factors: the lower density of probes on those arrays makes it more likely that they will completely miss a large number of CNVs; BAC-identified CNVs are often identified from altered hybridization intensity of just a single probe; the greater length of the individual probes increases the chance that results will be confounded by cross-hybridization to multiple genomic regions; BACs are subject to recombination, misidentification, and mismapping to the genome at any of the many steps between the creation of the BAC library in bacteria to the spotting of the BAC probes onto arrays. In addition, we have observed that 'home made' non-commercial microarrays rarely, if ever, approach the quality and reproducibility found in commercially produced arrays (data not shown). Regardless of the ultimate source of the lack of reproducibility of the BAC-aCGH-based CNV results, it is clear that those initial findings must be treated with caution.
Although most of the CNVs identified by Graubert are also identified by Cutler, the reverse is not true. Only 26% of the 2,094 individual CNVs mapped by Cutler fall within CNV loci as defined in Graubert (Cutler et al., 2007). We believe that this is due to a more conservative CNV-calling algorithm used by Graubert coupled with a compressed range of fold-change amplitudes that we have previously observed with the NimbleGen platform (data not shown and see below). It remains to be seen how the concordance would change if a more sensitive algorithm is used to analyze the data from Graubert.
Since Cutler is the most comprehensive analysis of mouse CNVs to date in terms of number of strains analyzed and has revealed the greatest number of CNVs, that body of data will be used for further discussion here, except where otherwise noted. That study identified 2,096 CNVs across 41 inbred mouse strains. This set of CNVs can be simplified by collapsing all overlapping CNVs between all strains into a set of 591 CNV loci, of which 224 loci contain CNVs from multiple mouse strains. The distribution of the remaining 367 singleton CNVs in terms of both length and amplitude is broadly similar to the 224 overlapping CNVs, though showing a statistically significant excess of lengths shorter than 10 kb (data not shown). Examining the full set of CNVs shows a mean length of 197 kb, close to the 323 kb mean length of CNVs identified in Graubert once those CNVs are mapped to the NCBI Build 36 of the mouse genome. The maximum size of detected CNVs on the autosomal chromosomes is in the 3 to 4 Mb range, although one CNV was found to extend over 8.6 Mb across the XMR locus on the X chromosome of Spret/EiJ. While there is no natural minimum limit to the length of CNVs, the density of probes on the microarrays used and the requirement of generally three or more adjacent perturbed probes imposes a technical minimum limit of approximately 36 bp. The smallest CNV actually detected is 62 bp long, although less than 0.3% of CNVs found were under 1 kb in length. This distribution of CNV lengths leads to an average of 10.1 Mb, or 0.38%, of the mouse genome being found within CNVs, with a maximum CNV content of 36.2 Mb, or 1.38%, found in Spret/EiJ.
The detected mouse CNVs are expected to have fold-change amplitudes that correspond to whole-number ratios. For example, a segmental duplication should have a fold change of 2 (a ratio of 2:1) and a segmental triplication should have a fold change of 3 (3:1), while a segmental deletion of a non-repeated sequence should have a negative-infinite fold change (0:1). This data should therefore be slightly easier to interpret than human CNV data where hemizygosity is an issue, and much easier to interpret than tumor CNV data where the DNA is isolated from samples that are generally contaminated with normal tissue resulting in a mixture of analyzed genomes. In fact, a histogram of the amplitudes of the identified CNVs matches closely what one would expect based on ratios of 1 to 3 diploid copies of genome segments, with the most prominent peaks corresponding to ratios of 0:1 (or –20-fold in this data set where extremely large and small fold-change values were truncated at 20-fold and –20-fold, respectively), 2:1, and 1:2 (Fig. 1). A similar analysis of the CNVs from Graubert shows a greatly compressed range of fold-change values, similar to what we have personally noted when working with NimbleGen arrays (data not shown), with fold-change amplitudes ranging only from –3.2-fold to 2.4-fold.
The amplitude histogram also reveals peaks that correspond to CNV amplitudes best explained as the whole number ratios 3:2, 2:3, 1:2, and 1:3. An example of a 1:2 'partial amplitude' deletion – as opposed to a 'full' deletion where there are no remaining copies of the locus – is found on chromosome 14 between positions 68206900 and 68418187 (genomic coordinates based on the Feb. 2006 version of the NCBI mouse genome assembly; Fig. 2). Plotting the fold-change values across this region shows a deletion with a mean fold-change of –2.2 across all strains except for C57BL/6J. As expected, probes for C57BL/6J are unchanged when compared to the reference data – other C57BL/6J samples – with a mean fold-change of 1.0. The most parsimonious explanation for this phenomenon is the presence of two diploid copies of this genomic region in C57BL/6J and only one copy in all the other strains tested. However, a sequence search using a sampling of 29 of the microarray probes in this region reveals only one genomic match per probe in the C57BL/6-based mouse genome sequence (data not shown). Therefore, this and the other 'partial amplitude' deletions likely reveal regions in the published mouse genome sequence where duplicated loci – including gene-rich, euchromatic regions such as the chromosome 14 locus shown here – have been misassembled as unique loci. The sequencing-based analysis in She supports this conclusion by also identifying a large amount of duplicated sequence in the C57BL/6J genome (She et al., 2008). Likely, both aCGH data and more sophisticated sequence analysis will be important information sources to help guide and correct sequence assemblies.
It is important to understand the nature of that portion of the genome which is found in CNVs, both to understand the processes that give rise to CNVs as well as their potential impacts on biology. Short tandem repeats that make up microsatellite sequences as well as repeated sequences on the order of several 10s of kbs in length have been observed to expand or contract in number through processes such as non-reciprocal recombination and gene conversion (Richard and Pâques, 2000; Read et al., 2004). Graubert explored whether segmental duplications, repeated regions of at least 90% identity, were associated with mouse CNVs and did, in fact, find such a correlation with 47.5% of 80 CNVs overlapping known regions of segmental duplication. Since our analysis of CNV amplitudes suggests that there exist previously unknown regions of high identity repeats, a true characterization of the association between CNVs and tandem segmental repeats is yet to be determined. An analysis of the association between repeats, including both simple (e.g. microsatellites) and complex (e.g. LINES, SINES) repeats, and the larger CNV set from Cutler, in contrast, shows significant enrichment of low-repeat-content DNA in deletions, 54% greater than would be expected if there were no association, coupled with a highly significant enrichment of high-repeat-content DNA outside of CNVs (Cutler et al., 2007). The reasons for this are unclear and warrant further analysis.
A potential source of false-positive deletion CNV calls from aCGH data may be the presence of SNPs in the sequences probed by the microarray oligos. However, the overall distribution of CNV amplitudes (Fig. 1) argues against this hypothesis, as does the lack of correlation between SNP content and CNV content in mouse strains (Cutler et al., 2007). This can also be tested by comparing the genomic locations of SNPs with the locations of CNVs. Interestingly, genomic regions of low SNP content (less than or equal to 0.5 known SNPs per kb) were very strongly enriched in both deletion CNVs (305% of the amount expected with no association) and amplification CNVs (239% of expected). As with the association with low-repeat content, the reasons for this SNP association are unclear and may be due to both technical and biological causes.
Perhaps of greatest interest is the association between genes and CNVs. Both intergenic regions and CpG-island-poor genomic regions are enriched in deletion CNVs with strong statistical significance (136% and 105% of expected, respectively) while genes and CpG islands are both enriched outside of CNVs. Amplification CNVs show enrichment of neither genes nor intergenic sequence and a modest enrichment of CpG-island-poor regions, while pseudogenes are strongly enriched in deletion CNVs. These data together suggest that the rate of formation of deletion CNVs is decreased in gene-rich regions and/or mice experience decreased viability or fitness when such events occur, unlike the situation with gene amplifications or pseudogene deletions which would generally be expected to be phenotypically neutral. Additionally, genes associated with known heritable diseases in humans can be found in amplification CNVs, but rarely in deletion CNVs (discussed in more detail below). Genes with up to two known paralogs in mice are significantly enriched (103%) outside of CNVs while genes with many paralogs (>5), and therefore with a biology or genomic structure that increases their propensity to have altered copy numbers, are indeed strongly enhanced in both deletion CNVs (353%) and amplification CNVs (230%).
The gene annotation provided by the Gene Ontology (GO) consortium (Ashburner et al., 2000) allows us to look for enrichment or exclusion of gene functions in CNVs. Both Cutler and Graubert find broadly similar results, with pheromone receptor and olfactory receptor-related functions, respectively, being strongly enriched in CNVs. These genes are found in large families marked by rapid evolution, as evidenced by the presence of many pseudogenes and a highly variable gene content between species, and are likely under reduced evolutionary pressure. Another gene function which is identified as CNV-enriched in both studies is defense response, including genes such as chemokine receptors, histocompatibility receptors, interferons, defensins, and killer cell-like receptors. As with pheromone and olfactory receptor genes, many of these defense response genes are found in large, variable families which, in the aseptic laboratory environment, are likely under reduced selective pressure. Cutler also identifies the related antigen binding, processing, and presentation functions enriched in both amplifications and deletions. These particular results must be viewed with caution, however, since they include genes involved in somatic-level rearrangements in lymphoid cells and thus may not represent germ-line CNVs. In contrast to the types of genes enriched in CNVs, many essential gene functions are found enriched outside of CNVs, including transcription, cell cycle, and protein folding functions.
We have already noted the inverse relationship between the distribution of CNVs and SNPs. One possible contributor to this phenomenon is a different phylogenic history of SNP-containing vs. CNV-containing genomic regions. For example, if two types of genetic changes arise with differing frequency, the one which arises less frequently may probe more deeply into history while the one which arises more frequently may be a shorter-term marker of a strain's lineage. Additionally, the randomness inherent in the chromosomal contributions of an organism's two parents coupled with the smaller numbers of CNVs as compared to SNPs makes it more likely that CNV content will fluctuate more across generations. To test whether there is a difference in the phylogenic history of SNPS and CNVs, we used the SplitsTree program (Huson, 1998) to generate a 'hybridization tree' based on both SNP and CNV data (Fig. 3). The SNP-based tree appears to be a good representation of the known history of these strains (Beck et al., 2000). The tree based on CNV data is broadly similar to the SNP tree, as can be seen by noting the colors of the major sections of the SNP tree carried over to the CNV tree. The main difference is that the common, interwoven sections of the trees which correspond to shared history among strains are much shorter in comparison to the strain-unique sections. This suggests a younger history for CNVs compared to SNPs.
These results are in line with recent studies which reported 'high rates of large-scale DNA copy number change' (Egan et al., 2007) and even CNV heterogeneity within C57BL/6J, which should be a completely inbred strain (Watkins-Chow and Pavan, 2008). In an attempt to calculate the rate of CNV formation in mice from the Cutler data, we selected 11 diverse strain pairs for which breeding history supported a common ancestor strain and with a good estimate for when the strains were created (Beck et al., 2000), including strains from the C57 group, the 129 group, the DBA group, and the FVB/SJL/SWB group. The ratio of CNV differences between the strains versus the number of years since the strain pairs have been separate was calculated for each strain pair (Fig. 4). These values fall within a reasonably narrow range of 0.17 to 0.56 CNVs/year of separation, with a mean of 0.37 CNVs/year. Using, as a very rough estimate, three generations per year (Egan et al., 2007), and taking into account that CNVs accumulate in both branches being compared (thus doubling the effective separation time), we can estimate approximately 0.6 new CNVs per mouse. This should be treated as a lower bound for several reasons, including the fact that we can only capture CNVs which are mappable on the C57BL/6 assembly, which are of sufficient size to detect above background given the aCGH array probe coverage, and which are not missed due to cycles of gain and loss or independent recurrence (Egan et al., 2007).
While it is expected that no essential genes would be found in homozygous deletion CNVs, genes which have significant effects on mouse phenotypes can and are found in both amplification and deletion CNVs. When murine genes overlapping CNVs were mapped to human homologs and compared to the OMIM database (OMIM 2007), 53 genes associated with human disease were identified (Supplementary Table 1, www.karger.com?doi=10.1159/000184721). Of these, only orthologs to three human disease genes were found in deletion CNVs (Cyp2c66, Abca4, and Serpina1 homologs). The human ortholog of the first of these, CYP2C66, which is deleted in CAST/EiJ, is one of the major enzymes responsible for the metabolism of cerivastatin, and was mutated in a patient that had rhabdomyolysis following treatment of hypercholesterolemia with cerivastatin (Ishikawa et al., 2004). The second gene we find deleted is Abca4 in the JF1/Ms strain, with most of its open reading frame absent. This CNV should lead to abnormal rod morphology in the eye as is seen with the laboratory knockout of this gene in 129S4/SvJae as well as in Stargardt disease, a human disease caused by mutations in ABCA4 (Wang et al., 2007). Finally, we observe deletions of members of the Serpina1 family in AKR/J, LP/J, I/LnJ, RIIIS/J, SPRET/EiJ and Balb/cJ mice. SERPINA1, also known as alpha-1 antitrypsin, is a serine proteinase inhibitor. One major physiological function of alpha-1 antitrypsin is the protection of the lower respiratory tract against proteolytic destruction by human leukocyte elastase (HLE). In humans, a hereditary deficiency of alpha-1 antitrypsin is associated with an increased risk of developing emphysema and chronic obstructive pulmonary disease (Crystal, 1990). AKR/J has been shown to be 'supersusceptible' to cigarette smoke- induced emphysema as compared to NZWLac/J, C57BL6/J, A/J, and SJ/L strains (Guerassimov et al., 2004). It is of note that these four strains have a full complement of Serpina1 genes. The AKR/J mouse has been found to have slightly lower lung elastance and slightly higher inflammation than the pallid mouse, a C57BL6/J variant strain with very low level of alpha-1 anti-trypsin (de Santi et al., 1995).
Other deletion CNVs which have potential phenotypic impacts are also seen. For example, RIIIS/J, PWK/PhJ, and WSB/EiJ contain deletions in the Ifi204 gene. This gene or its human homolog IFI16 have been implicated in the development of mononuclear phagocytes (Bourette and Mouchiroud, 2008), modulation of ras activity (Ding and Lengyel, 2008), DNA damage and checkpoint control (Ouchi and Ouchi, 2008), as well as cancer and autoimmune disease (Choubey et al., 2008). The deletion CNVs identified cover a region annotated as intronic, but which overlaps sequenced mRNAs as well as spliced ESTs and therefore likely does contain exons.
The remaining 40 genesassociated with human genetic disorders were all found in regions of copy-number increase as compared to C57BL6/J. Although these CNVs may have phenotypic effects, their presence is less likely to result in overt disease as compared to missing genes. Since this summary only covers CNVs that overlap with protein transcriptional reading frames and ignores those which potentially span transcriptional regulatory regions and non protein-coding transcripts, it is only a starting point in understanding the involvement of CNVs in disease etiology in the mouse.
As discussed earlier, one notable bias in the overlap between genes and CNVs is the preponderance of CNVs covering genes related to defense against pathogens. Gene copy number in killer cell lectin-like receptor (Klr) genes is highly variable (Anderson et al., 2005; Cutler et al., 2007; Graubert et al., 2007) and can explain differences in susceptibility to cytomegalovirus infection (Lee et al., 2001) as well as response to tumor (Nakamura et al., 1999). A number of mouse strains have complete deletions of multiple members of this gene family (Supplementary Table 1), which are concentrated in a locus on chromosome 6 (Fig. 5). Because this variation may affect the ability of these strains to respond to challenges to the innate immune system, researchers studying these processes should consider evaluating their data in light of this CNV data. Although informative, we know that the aCGH analysis is underestimating the CNV diversity of these genes. When directly sequenced, the Balb/cJ mouse was found to not only have the deletion CNVs noted here, but also the presence of additional genes absent from the C57BL6/J genome and therefore not probed for by the arrays (Anderson et al., 2005). This suggests that sequencing of additional strains could lead to the discovery of additional members of this family.
In many strains (SPRET/EiJ, MOLF/EiJ, RIIIS/J, MSM/Ms, and JF1/Ms), we observe high-level increases in copy numbers of the Itln1 gene compared to C57BL6/J. The copy number is likely to be greater than 20 in the RIIIS/J strain. The protein product of this gene (known by various names including intelectin, lactoferrin receptor, and omentin) has been implicated in binding lactoferrin and iron absorption (Suzuki and Lönnerdal, 2004), associated with obesity (Yang et al., 2006), and involved in response to pathogenic infection in the gut (Wrackmeyer et al., 2006). This gene has a paralog (Itln2) in some mouse strains (Pemberton et al., 2004), but which is not present in C57BL6/J so therefore not on the CGH microarrays used by Cutler and Graubert. Orthologs to both genes, Itln1 and Itln2, are present in humans. Interpretations of murine in vivo studies of either protein, or phenotypes related to known functions of these proteins, must take into account this copy-number information.
A locus containing three Raet1 genes (retinoic acid induced early transcript) shows copy number changes in many strains. These major histocompatibility complex (MHC) class I-related gene products have been shown to bind to the C-type lectin receptor, NKG2D (also known as KLRK1), present on natural killer cells as well as certain subsets of T-cells (Lanier, 2005). Because this interaction is involved in immunosurveillance of tumor cells (Strid et al., 2008), models which involve de novo tumor formation (either spontaneous or induced) can be affected by the absence of some or all of these genes.
The Tlr7 gene (as well as nearby genes) on the most distal part of the X chromosome has been shown to be duplicated in SB/Le mice and confers increased severity of systemic lupus erythematosus (SLE) in male mice. By fluorescence in situ hybridization (FISH) mapping, this duplication has been shown to exist on the Y chromosome explaining the nature of this Y-linked autoimmune accelerator (Yaa) locus (Pisitkun et al., 2006). Analysis of the Cutler CNV data suggests that the FVB/Ntac strain also contains a duplication of this locus. While it is unknown whether this duplication exists on the Y chromosome in FVB/Ntac, it is likely to have the same phenotypic consequences.
Another crucial component of the innate immune response is the defensin family (Selsted and Ouellette, 2005). This family of cysteine-rich peptides is expressed in multiple different tissue types including leukocytes and intestinal crypt cells. In mice, α-defensin expression is absent in leukocytes, but several α-defensins can be expressed in the Paneth cells of intestinal crypts. Additionally, more than 50 β-defensin genes have been found in mice. The host response to microbes in the intestinal tract appears to be a vital aspect of the pathogenesis of inflammatory bowel disease (IBD, reviewed in Wang et al., 2007). It has been shown that copy number polymorphisms are prevalent in the β-defensin genes, and reduced copy numbers predispose people to Crohn's disease (Fellermann et al., 2006). In nearly half of the strains we examined, copy number variations in several of the β-defensins as well as complete loss of the defensin-related cryptdins were found. Unless similar genes exist in these strains to compensate for the missing functions of these defensin-related cryptdins, one would expect that the lack of antimicrobial peptides expressed in the intestinal crypts would predispose these strains to microbial invasion of the mucosa and IBD. Although it is clear that IBD is a multigenic disease with multiple environmental influences, study of these pathologies should consider the copy number changes seen in our mouse models.
The high degree of variation in genes involved in the innate immune response could be due to the way in which laboratory strains are bred and maintained. Over time, gene duplication coupled with diversification leads to the development and expansion of gene families, such as the killer cell lectin-like receptors and defensins discussed above. Each family member may serve overlapping as well as unique functions compared to the other family members. As laboratory strains are largely protected from immune challenges that they might face in the wild, the selective pressure to maintain the diversity of these families could decrease and individual genes can be lost from populations and then that loss will be fixed through inbreeding.
Inbred mouse strains have been an important model for studying complex biological phenotypes since the beginning of the 20th century. The power of this disease model was greatly enhanced with the sequencing of the mouse genome in 2002 (Mouse Genome Sequencing Consortium, 2002). However, a static genome sequence belies the actual dynamic state of the genome as revealed by recent CNV analyses (Li et al., 2004; Snijders et al., 2005; Cutler et al., 2007; Egan et al., 2007; Graubert et al., 2007; Watkins-Chow and Pavan, 2008). Furthermore, this CNV data points to deficiencies in the published genome sequence, specifically the misassembly of some repeated genomic regions and the likelihood that sequences present in other mouse strains are absent from the sequence based on the canonical C57BL/6 strain. Researchers performing reverse genetics in mice, such as creating knockout or mutant strains, need to be aware of the copy-number status of their gene target. The phenotype of a knockout mouse may be very misleading, for example, when a wild-type repeated copy of the targeted gene still exists. Researchers engaged in forward genetics analyses, such as WGA, can also put this knowledge of the mouse CNV landscape to work. This has been demonstrated recently, where an amplification at the glucagon-like peptide-1 receptor locus was found to be linked to food intake levels across an assortment of mouse strains (Cutler et al., 2007). Finally, since inbred strains, in large part, reflect the genomic diversity of the individual mice that served as founders for the strains, their genomes reflect a snapshot in time of genome evolution. This developing picture of the mouse CNV landscape provides insight into the forces that shape genomes in both mice and humans.
The data used in the analyses shown here all comes from Cutler et al. (2007), unless otherwise noted. Numbers of disparate CNV loci were calculated by 'collapsing' CNVs from all strains which either overlapped by at least 8 kb or where one CNV was less than 8 kb and was entirely contained within a larger CNV. Smoothed fold-change histograms were generated using the R function smooth.spline with 38 degrees of freedom on a histogram with 70 breaks. Mouse phylogenetic trees were generated using the SplitsTree4 program (Huson, 1998). For the CNV-based tree, the collapsed CNV set was used. The CNVs were converted into a pseudo-nucleotide-sequence format based on this conversion key: CNV amplitude ≤–10x → 'AAA', ≤–2.5x → 'AAC', <–1x → 'ACC', ≥10x → 'TTT', ≥2.5x → 'TTC', >1x → 'TCC', no CNV → 'CCC'. SNPs were also converted into a pseudo-sequence file by extracting the 8,315 SNPs with the greatest strain coverage from the SNP database found at http://phenome.jax.org/pub-cgi/phenome/mpdcgi?rtn=snps/download. Ungenotyped SNPs were replaced with 'N' and conflicting SNP genotypes in the downloaded data set were replaced with the appropriate ambiguity codes. These pseudo-sequences were then imported into SplitsTree4 as FASTA files. The settings used were, for the distance metric: 'Uncorrected_P' with 'AverageStates' for ambiguous positions and 'Normalize' set to On; for the network type, 'HybridizationNetwork' was selected.
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Accepted: June 17, 2008
Published online: March 11, 2009
Issue release date: March 2009
Number of Print Pages: 10
Number of Figures: 5
Number of Tables: 0
ISSN: 1424-8581 (Print)
eISSN: 1424-859X (Online)
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Eli Young Band gets up to speed for Kenny Chesney opener
Opening up stadiums for Kenny Chesney's No Shoes Nation tour was a bit of an adjustment, said James Young, guitarist for the Texas-based Eli Young Band.
"It would have been nice if we could have actually warmed up with an arena or amphitheater," Young said of playing the tour kick-off at Raymond James Stadium in Tampa, Fla., in March. »Read Full Article
Smart staging makes good 'Company' at Theatre Unchained
Putting aside the juicy tax breaks, is marriage worth it?
That's the question on the table in Stephen Sondheim's "Company," and to paraphrase from another Sondheim show, the answers in this great master's work are never easy. »Read Full Article
Top of the Charts: Pharrell Williams celebrates seventh week on top
Seven is the lucky number all around this week, as Pharrell Williams celebrates his seventh consecutive week on top of the Billboard Hot 100 and all of the top seven positions remained locked in position this week. Watch the video for Williams' big hit single below and then let's reset our decades and travel way back in time to see the songs that were the biggest in the land this week in 2000, 1990, 1980 and 1970.
No. 1 hit April 14, 2014 »Read Full Blog Post
On Tap Today: Milwaukee Day, Morgan Freeman and the Brewers
Hey, it's Milwaukee Day. And on this fine 4/14, local band Decibully reunites.
Time flies when you're watching movies. Marcus Theatres celebrate the 20th anniversary of "Shawshank Redemption" with showings beginning today. »Read Full Blog Post

subset: commoncrawl


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Actor Joaquin Cordero died at his home in the Mexican capital, the secretary of the ANDA actors guild told Efe. He was 89.
Cordero was "one of the most beloved people in ANDA," Amparo Garrido said.
While Garrido did not specify a cause of death, Mexican media suggested the actor fell into a depression after the death seven months ago of his wife, Alma.
Cordero was also recovering from an embolism.
Born Aug. 16, 1923, in the central city of Puebla, Cordero was among the last surviving figures from the Golden Age of Mexican cinema, when he appeared alongside artists such as Mario Moreno "Cantinflas," Pedro Infante and Carmen Montejo.
Cordero made more than 200 films over seven decades and was later a featured performer in telenovelas, including "Destilando amor," "Fuego en la Sangre" and "La Madrastra." EFE


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Please upload a photo that has a resolution of at least 0 dpi at its print size and is in one of the following file formats: . The file must not exceed 10MB. Beau-coup will not crop or retouch your photo and is not responsible for the quality of uploaded files. By uploading an image, you agree to our terms and conditions.
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Comments about Beau-coup Acrylic Heart Shaped Photo Cake Topper:
I cannot thank the staff enough. They were helpful all the way.
Bottom Line Yes, I would recommend this to a friend
Acrylic heart shaped topper
By Danielle
from Ontario, Canada
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Comments about Beau-coup Acrylic Heart Shaped Photo Cake Topper:
I purchased this cake topper as I really liked the idea of having the both of us represented on our wedding cake. When I took it to my baker, she asked how it was going to hold on top of the cake - I would suggest adding a base or give us ideas on how to make it stay put :-)
I have had a great service with my previous and first order with Beau-coup - very professional service - thank you so much!
Bottom Line Yes, I would recommend this to a friend
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in-laws 40th anniversary
By Glo727
from Mira Loma, CA
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Comments about Beau-coup Acrylic Heart Shaped Photo Cake Topper:
My in-laws are renewing their vows. My sister-in-law and I got this for them as a gift for their cake that I will be making. We will put on one side a picture of their wedding day and on the other side a picture of them now. I love the frame. The only thing is that I wish it was a little bit bigger... That is why I gave it 4 stars.
Bottom Line Yes, I would recommend this to a friend
(0 of 1 customers found this review helpful)
Simple yet totally swoon-worthy, this acrylic heart-shaped photo cake topper is an easy way to add a perfectly personal touch to your wedding cake.
Display two photos back-to-back inside this clear, heart-shaped topper and slip atop your cake for maximum impact. Measures 4"L x 4"W x 0.5"H.
Want more one-of-a-kind topper options? Take a look at our personalized round wedding cake topper, comical wedding cake toppers and brushed metal monogram cake toppers.
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Christine D. Long pleaded not guilty after her indictment last year on 10 counts of cruelty to children and five counts of failure to provide an education for her school-age children. She is expected to change her plea to guilty in Burke County Superior Court and receive her sentence today.
Her husband, Jeremy Long, pleaded guilty in June. He was sentenced to three years in prison followed by 10 years on probation.
Mrs. Long blamed the children's condition on her husband and further accused him of being an abusive man who used violence, sexual abuse and starvation to keep the family prisoners in the hovel where they were found, according to her petition for divorce.
Read more from Thursday's report


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DATS provides door-to-door specialized transit for registered pre-booked passengers 16 years of age or older who have a severe physical or cognitive disability and are unable to take conventional public transit for all or some trips. DATS takes reservation bookings three days in advance of travel, two days in advance of travel, and the day before until noon. Subscription bookings are available for trips required on a regular basis.
Over the summer of 2015, DATS led collaborative workshops, exercises and surveys with customers, caregivers and staff to determine a new vision, mission and values.
Vision: To provide industry leading and sustainable specialized transportation solutions that connect people, resources, and communities.
Mission: To provide customer focused, safe, and reliable door-to-door shared ride services for those who are unable to use conventional transit.
Values: Safety, Respect, Customer Service, Compassion, Teamwork
All passengers must be registered with DATS before booking trips. For more information, call the DATS Customer Care Centre.
- Trip Booking Request and Cancellation Forms
- DATS Hours of Service and Booking Procedures
- DATS Fares
- DATS Advisory Group
- 360° Tour of a DATS Van


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Home > Indexes > OKI > 43381717
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UPC Code: 051851353623
Provantage Code: OKIC04H
Condition: Factory New
Printing > Printer / Copier Consumables > Printing Drums
Yellow Image Drum for C6100 Series Printers
You may qualify for additional discounts if you purchase at least 10 units of OKI hardware products.
Shop with confidence! Provantage is an authorized OKI dealer.


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Gamer Tonight
Every episode features a different video game stereotype!
Neo Bender
The very popular Futurama / Matrix crossover series, with amazing 3D Flash!
Minecraft: TNA
Minecraft N00BS go on hilarious adventures.
congrats mrs head its....a giraph!!
going to make more?
going to make more balloon head freds?
do you fuck a balloon?
One of your best
I really like this one and think it is among the most memorable you have made. As stated somewhere else, you don't have many women in these cartoons, but you do draw them well when you can. While it may have been relatively short, it got its delivery off quite well. It was also funny to see the woman give birth to the balloon toy. What's really nice is how earnestly the characters look at each other, especially the balloon guy. May I wish you a very happy new year and many more to come!
balloon, damn you balloon
He hate his baby and his wife, he wants only sexy and scat bysexual orgyes lolololololololololololololololololol olololololoolloololollolololololololo lolololroflroflroflroflroflroflroflor lfroflforlfoflofffr


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What we're up to
- Living your possibilities
- Volunteer trip to South America inspired by mentoring
- more...
- Big Brothers Big Sisters Goes Back to School
- After a long summer break, mentoring matches reunite in the academic setting
- more...
- Big Brothers Big Sisters Has Volunteer Big Sisters Waiting
- Big Brothers Big Sisters of Middle Tennessee is a free mentoring program serving children throughout the Nashville and Middle Tennessee area. Currently, the nonprofit organization has several volunteer Big Sisters waiting to help mentor a child - with a shortage of girls requesting an additional positive adult role model in their lives.
- more...
Our Mission
Big Brothers Big Sisters is providing children facing adversity with strong and enduring, professionally supported one-to-one relationships that change their lives for the better, forever.
Stay in the Know
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Have some fun and help start something today for children throughout Middle Tennessee.Read on >


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Environmentalists see a fight looming in City Council over a bit of business left over from last year's zoning code reform - a bill that would determine how close something can be built to the city's rivers and streams.
Legislation introduced in September would create a 50-foot buffer, or "setback," around those bodies of water - less than the 100 feet environmental advocates preferred, but a number they saw as a compromise with builders.
Now environmentalists fear Council will try to reduce the setback on the city's streams - basically, everything except the Schuylkill and Delaware Rivers - to 25 feet, a distance they say could harm already polluted waterways.


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Personaly, i love sonic. He's fast, agile, and not to mention, fast. however, i heard that people think sonic is a horrible character. I think he's just too fast for some people. yes, he is pretty weak, but he can rack up combos fast. and his final smash is pretty awesome too.
you should post this in the smash character disscussion section.
i like sonic too but his b, fowardb and dash fowardA are all becoming a ball.
he needs bigger hits.


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Singapore PM
Nov 29, 2015
Singapore Prime Minister Lee pays tribute to Sikh community
Prime Minister Lee Hsien Loong has paid tribute to Singapore's Sikh community, saying the government will continue to support them in many ways. #India #NewsTracker #Singapore #Indian diaspora #Sikhs #Singapore PM #SIngapore turns 50
Jul 13, 2012
India can grow 8-9% with reforms: Singapore PM
Hailing India's efforts since economic liberalisation in 1991, he said the country needs to capitalise on its large human capital and build on the success in the information technology sector. #India #TheySaidIt #Lee Hsien Loong #Singapore PM
Jul 12, 2012
Lee invites India to join ASEAN's new economic partnership
Singapore Prime Minister Lee Hsien Loong today pitched for India's active participation in ASEAN's new Regional Comprehensive Economic Partnership to lend stability in the region. #NewsTracker #Asean #Singapore PM
Jul 10, 2012
Singapore PM Lee's India visit to focus on investment, trade
Singapore Prime Minister Lee Hsien Loong will make a two-day state visit to India from tomorrow, seeking to boost the already strong and multifaceted bilateral ties which has seen growing trade and investments. #India #NewsTracker #Lee Hsien Loong #Singapore PM


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This book ranks easily among my top five audiobooks of all time. It was simply a fantastic listen.
Doc was my favorite because he proved just how complex yet simple friendship and life can be viewed al at the same time through the time he spent with the lead character.
No this is my first listen to one of his performances.
It would have to be Doc, for the same reasons he is my favorite character.
I laughed, cried and at times was outraged while listening to this book. It is truly an experience. It is definitely one of the best performances that I have had the pleasure to use a few hours of my life to listen to.
I most enjoy listening to contemporary fiction with well-developed characters.
"My first Courtney but certainly won't be my last"
A story of hardship and struggle and how a young boy navigates through it with the help of a very special mentor.
I bought this audiobook based on the reviews and knowing nothing of the author. I am not a fan of boxing so I was surprised at how entirely this book drew me in. It was engaging and uplifting... just what I love in a book.
The narrator did a great job.
"interesting and especially well read"
the reader was/is fantastic
I have listened and liked very much the other books of the same "couple"
Funny that actually one does not know what finally happened to the main character
Traveler. Artist. Dreamer.
"Am I the only one that thinks this was a let down?"
Peekay is a remarkable boy who is so remarkable in fact that it is practically unbelievable. Mr Courtenay focuses on how remarkable this boy is the whole time and then ends it with a very unremarkable moment. It was such a let down and it just does't make sense... What about being the tadpole angel or being the welterweight champion of the world? Why did he put so much emphasis on these two aspects and then end it with a silly confrontation? It's as if there was a build up for nothing. The end came straight out of left field and seems inconsistent with the direction the story was headed.
As much as I think the story was a let down, there are 3 things that I really liked about The Power of One:
1. The character development was "wunderbar, absolutal"
2. It had important messages about life scattered throughout
3. Humphrey Bower was an excellent narrator
Loved this book! I can't believe I just now have discovered Bryce Courtenay. Wonderful plot, characters, descriptions. Listened practically non-stop. Mr. Bower has a great voice and adds to the enjoyment of this terrific story.
Try this!
I was not aware of Bryce Courtenay until recently. This is a really good story made great by the narrator Bower. The story, the voices and the historical perspective reminded me of Shantaram. Bower really is terrific.
"Brilliant partnership of author and narrator"
Definitely one of my top ten
I was born in South Africa, and lived there for 35 years, so I could appreciate the language, the humor, and sadly, the immense cruelty of the time.
I first heard Humphrey Bower after listening to Shantaram, and he was brilliant. I had read The Power of One many years ago, and decided to download the audible version. I was not disappointed. Bower brought Peekay and the Professor to life, and the story had a depth that I did not experience when reading it. I immediately downloaded Tandia which is a continuation of Peekay's story, and can't wait to start that.
You may want to consider acquiring another book by the same author called "Whitethorn". If is is read by Humphrey Bower, I know it will make a great listen
It's the perfect title
"A must read for all"
The yarn, the characters, the pace of action...nothing I didn't love....
Grandpa Chook - loyal, clever, just wonderful way to tell of hope and willpower.
Humphrey characterises everyone brilliantly. Not fair to choose.
Gideon - then he could stay over!
"Courtenay is a Master Storyteller!"
Sometimes you come across a gifted writer. Sometimes you come across a gifted storyteller. When you come across both of these attributes in one person, then you have a legend. I am just sorry that I was introduced to Courtenay by way of his obituary. The Power of One was his first book; one of many that I will read.
A bright young boy sent off to boarding school at the age of 5, after the death of his father {and his mother subsequently committed to a "nervous breakdown" institution}, has to find his way in a very socially complicated and racially divided society. He soon discovers his dream is to become a champion boxer, but only after first conquering many humiliations and fears. The bullies that nearly break him in boarding school leave open an emotional sore, which is soon mended by people who, by happenstance, enter his life and become integral forces in giving birth to a tenacious champion of both body and soul. I am listening to this book rather than reading it. The narrator, Humphrey Bower, does an outstanding portrayal of every character in the book. I highly recommend the audible, if you want to be truly entertained.
Best Listen Ever! I so thoroughly enjoyed this book. Not only is the story amazing, the characters are brought to life by the work of Humphrey Bower. Sometimes I find it difficult to remember that only one person is narrating the book! And yes, it's about boxing. I don't care for boxing, but it's such a vital part of the story that I'm enjoying every part of it!
Other than PK - Doc! Such a quiet source of wisdom and encouragement!
He manages to become every character in the book seamlessly, including the variety of accents. I always believe in the character who has the voice at any one time!


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Items Tagged with 'best'
What's on your personal list?
Everybody loves a list. Over the past several days, grbj.com has reported on West Michigan cities appearing on several complimentary lists for business and lifestyle. Read More
It's not really failure — it's failure to do your best
I'm often asked, "Why do salespeople fail?" The answer is: They don't fail. Read More


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KIBBUTZ KETURA, Israel - A $15 million investment bought Siemens a 40 percent interest in the Arava Power Company, Israel's leading developer of solar power.
"This investment is another consequential step in further strengthening our green and sustainable technologies," Siemens AG President and CEO Peter Löscher said.
"Thanks to its intensive sunshine and steadily growing demand for energy, Israel is an ideal location for further developing our solar business," Löscher said.
Siemens' investment - the largest ever foreign investment in an Israeli solar power company - will allow Arava to build the nation's first commercial solar plant, in the desert region south of the Dead Sea.
Johannes Schmidt, CEO of Siemens Financial Services, said it was "the most comprehensive investment in an Israeli solar power company so far."
"Because of its early entry and wide activities in the field, Arava Power has become the leading solar power company in Israel," Schmidt said.
"Siemens will support the local solar projects with our variety of technologies, professional expertise and funding capabilities," he said.
Israel hopes that renewable sources will provide 10 percent of the nation's energy needs by 2020.
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Are you seeking answers in life? Are you hurting? Are you facing a difficult situation?
Find peace with God, discover more about God or send us your prayer request.
Call The 700 Club Prayer Center at 1 (800) 823-6053, 24 hours a day.
A caring friend will be there to pray with you in your time of need.
CBN News is a national/international, nonprofit news organization that provides programming by cable, satellite, and the Internet, 24-hours a day. Follow us on Facebook, Twitter, and Google+.
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The Terrorists Funded by the WestBassam Tawil
The New Israel Fund and those who support it are helping terroristsRonn Torossian
Column one: Fighting fire with fireCaroline B. Glick
Op-Ed: There is no 'radical Islam' and there is no 'moderate Islam'Dr. Mordechai Kedar
'Narrow' escapesRuthie Blum
Why Palestinians Do Not Want Cameras on the Temple MountKhaled Abu Toameh
Social Media Killed My FatherYoni Kempinski and Ari Soffer
Iran's New Palestinian Terror Group: Al SabireenKhaled Abu Toameh


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Travel Advice
Lay-by your airfare: latest price war strategy
- From: The Courier-Mail
- July 31, 2007
QANTAS has thrown the latest punch in Australia's air fare wars, not by slashing prices but by offering deferred payment on a range of its flights.
The deal applies to most international fares and fully flexible or business class domestic fares.
It will allow those booking online to virtually lay-by their fares, secure in the knowledge their preferred flight is reserved.
Qantas Group general sales manager Rob Gurney said passengers would be charged a holding fee of $25 a passenger.
But how long you have to pay the flight depends on the fare type.
If you want to pay $14,840 for a business class return fare from Brisbane to Los Angeles in May next year, returning a month later, you have until midnight May 29, 2008 to make full payment.
Opt for a $2560 Red e-Deal on the same dates and you only have until Sunday to pay the balance.
A $860 fully-flexible return fare from Brisbane to Sydney on August 13, returning a week later, would entitle a customer to delay payment until August 12.
Qantas's low-cost carrier Jetstar also has been aggressive in the face of cheap fares from Tiger Airways, offering a lowest price guarantee.
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Use our girl baby shower tablecovers to make your baby girl shower tableware really stand out. Use solid or patterned tablecovers to bring all your table decorations together and to make clean up a snap! Choose a style that matches your tableware and decor.
For your shower, our girl baby shower table covers set the tone for your tableware. Choose a tableware ensemble from our baby girl shower tableware themes. Cover your table with a coordinating solid or print table cover. Add a baby shower centerpiece for a beautiful focal point. Finish it all off with sweet baby girl confetti!


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A wall has a negative charge distribution producing a uniformhorizontal electric field. A small plastic ball of mass 0.01 kg,carrying a charge of -80.0 C, is suspended by an uncharged, nonconductingthread 0.30 m long. The thread is attached to the wall and the ballhangs in equilibrium, as shown above, in the electric andgravitational fields. The electric force on the ball has amagnitude of 0.032 N.
(a) On the diagram below, draw and label the forces acting on theball.
(b) Calculate the magnitude of the electric field at the ball'slocation due to the charged wall, and state its direction relativeto the coordinate axes shown.
(c) Determine the perpendicular distance from the wall to thecenter of the ball.
(d) The string is now cut.
i. Calculate the magnitude of the resultingacceleration of the ball, and state its direction relative to thecoordinate axes shown.
ii. Describe the resulting path of theball.


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Photographing the Eli Young Band
Lesson 1: Introductionwith Jeremy Cowart
Duration: 1 hour 29 minutes
Photographers and country music fans alike are in for a special treat when Jeremy Cowart invites Matt Kloskowski and KelbyTraining.com to join him on a commercial shoot, photographing country recording artists from Universal's Republic Nashville label - The Eli Young Band. You'll get an up-close and personal look at the guys behind the band as Jeremy takes the photos for their latest album, Life at Best, which just debuted at #6 on the Billboard 200 chart. You'll also learn some killer commercial lighting setups and see what it takes to produce a shoot for a major-label release as Jeremy sets up in a super cool (and freezing cold) Tennessee prop warehouse. Then, back in the KelbyOne.com studios, Jeremy performs the actual editing the client asks for so you can see exactly what he did before these pictures were put to use. See the final shots the label used and learn more about the Eli Young Band at http://www.eliyoungband.com
No course objectives yet
No materials available for this course
Jeremy Cowart
Jeremy Cowart is a professional photographer from Nashville, Tennessee. Beginning his photography career in 2005, Jeremy quickly became a respected artistic voice in the industry. Having shot numerous musicians, entertainers and celebrities, Jeremy is also the founder of Help-Portrait Movement, a worldwide movement of photographers giving free portraits to those less fortunate. As his list of clients continues to grow, so does Jeremy's desire to improve, share, teach, and give back to those around him.


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Very important Apremilast discussion @ Piper conf discussing how Apremilast efficacy continues to improve the longer patients on drug vs the plateau seen with biologics.....
: ...or continued benefit versus a plateau effect that's typically seen with the biologics.
: Yeah.
: Is that something that's continuing? Does that really point to potential benefit that at some point gets to a biologic or potentially better than a biologic?
: Yeah. That – it's a very interesting point, Ian. Thanks for bringing that up. So, the ACR20s that we saw at 16 weeks continued to improve. There was actually a 50% improvement from week 16 to week 24 in the ACR20s. And you don't see that in the biologics. Usually with biologics, the patient will plateau and they stay at that efficacy level continuously. We did see this increase.
Now we're continuing to file these patients out, and the studies will continue to run through 52 weeks and then we're actually going to follow for a couple of years for safety. So, we will see if these – if this benefit continues to increase. But it does seem to be a very unique feature of apremilast that the patients as they're on drug longer continue to improve. And we'll see where they ultimately do plateau or how long they continue to improve. But that's a very positive aspect of the drug, especially when you have a continuous chronic therapy twice a day. Patients are much more likely to adhere to that therapy if they continue to improve which is what we did see.


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-Victoria Jones- Unisex black background cotton ramie blend pullover longsleeve Ugly Christmas Sweater, oversized turtleneck collar featuring three holly leaves with red bow and red, green, white, and silver beads. Plain on back. Sleeves have scattered white and silver beads, the bottom is lined with green holly leaves and red, white, and silver beads. (Made for a woman but would work on a guy, just scrunch up sleeves if they are too short)
Fits 38Chest (Mens Small) or 38Bust (Womens Large), 29Sleeve, 28Inches from Shoulder to Hem
Good Condition
1.59 lbs
Price for both: $47.00
Buy Both


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Originally Posted by Kriblover
Nice tank. What's that yellow thing in the first pic on the third set of pics?
Its a yellow goby he loves his home haha, everything growing great looking even better now will post more pics soon thanks for the comments everyone
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Two stacked aquariums available for local pick up
Fully equipped with lights, covers, penguin ...
I am in the label printing business and I have a
salt water aquarium. I cam up with these labels
and ...
Welcome to Aquarium Home Farm Center, a
professional breeder of the Dragon Fish, the Asian
Red Arowa ...


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WAREHOUSE 13. 9 tonight, Syfy.
EUREKA. 9 p.m. Friday, Syfy.
BEFORE WE make too much fun of the Russians for going to such trouble to infiltrate American suburbia - could another "Real Housewives" edition be far behind? - consider what they may have learned just for the price of their sleeper agents' cable bills.
For one thing, that U.S. tax dollars are spent for things we'll never hear discussed on C-SPAN.
Take Syfy's "Warehouse 13," which returns tonight for Season 2.
The rebranded NBC Universal network's most successful series ever - its first season averaged 4.1 million viewers - it focuses on a couple of U.S. Secret Service agents, Myka Bering (Joanne Kelly) and Pete Lattimer (Eddie McClintock), whose job isn't guarding the president or the U.S. Treasury but rounding up supernatural "artifacts" for a storage facility in South Dakota whose eccentric caretaker, Artie, is played by Saul Rubinek.


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INNSIDE Bremen4
Price FromView Rooms
This hotel doesn't have any reviews yet.
About the hotel
This hotel is located near Bremen city centre and is only a few minutes by public transport from the many amenities of the city such as restaurants, shops, bars and further entertainment venues. Bremen Airport is about 7 km from the hotel. The hotel comprises a total of 177 rooms set over three floors, of which 15 are suites.
Facilities on offer in this city hotel include an elegant foyer with 24-hour reception, express checkout, a hotel safe, a cloakroom, a currency exchange counter, a café and lifts. A cosy bistro-bar and an air-conditioned restaurant (with non-smoking area and highchairs for children) cater to the culinary desires of guests. A selection of rooms are available for use by business guests, which are suitable for conferences, congresses and banquets. Room and laundry services complete the exclusive offerings (charges apply). Guests may make use of the WLAN access point and there are on-site parking spaces for those arriving by car (charges apply). The pleasantly fitted rooms all come with an en suite bathroom with hairdryer, a direct dial telephone, satellite/ cable TV and a radio. Further features include a minibar/ fridge, central heating, an Internet connection, a double or king-size bed and carpeted floors. All rooms and bathrooms are disability-friendly. There is a spa centre with sauna, Jacuzzi, sun terrace and solarium available to guests. Sports enthusiasts may like to try a workout in the gym. Guests may select their breakfast from a buffet. Lunch and evening meals can be taken à la carte. The kitchen also accepts suggestions for particular dishes. Tram No 3 goes from the city center pedestrian zone to the Hotel in 15 minutes. Autobahn A27 is 3kms away, good access to A1 and A28
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Facilities Show/hide
- Main
- 24 Hour Reception , Safety Deposit Box , 24 Hour Room Service , Currency Exchange , Lift to all floors , Public Air Conditioning
- Leisure
- Gymnasium
- Dining
- Breakfast to go
Useful Info Show/hide
- From 14:00 hours
- Until 11:00 hours
- 09/02/2012 to 31/12/2021 - new city tax for leisure travellers only to follow in 2013 to be paid in addition to room rate at the Hotel directly The price shown is based on the occupancy and dates you have searched for. Prices and availability are subject to change based on the arrival date, number of nights and occupancy searched for. All prices are inclusive of all local taxes.
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Dwight King picked up his second goal in four games during Thursday's 3-2 win over Montreal.
Dwight King tipped home the eventual winner in Thursday's 2-1 win over Edmonton.
Dwight King picked a goal and an assist in 4-3 OT loss to Colorado on Wednesday.
Dwight King (foot) was activated from the IL on Tuesday December 29.
Dwight King (foot) practiced with the Kings for a second straight day Tuesday, but he is still on injured reserve.
Dwight King (foot) has started skating on his own.
Dwight King (foot) isn't expected to resume skating for another two weeks.
Dwight King is still a couple of weeks away from returning to action from a broken bone in his foot.
Dwight King (foot) is still on crutches.
Dwight King, who was hurt during the preseason, broke a bone in his foot.
Dwight King (lower body) has been placed on injured reserve.
Dwight King was able to tally his 13th goal of the season Monday night versus the Vancouver Canucks.
Dwight King picked up a goal for the Kings in their 4-1 loss to the Wild on Saturday night.
Dwight King practiced on a line with Marian Gaborik and Anze Kopitar on Monday.


text:
Total # Posts: 21,982
You too! Sra
November 26, 2014
14. recogido is a past participle and must follow a form of haber OR it is an adjective, following either ser or estar. = el jefe recogió dinero 16. El tono es un poco triste?...pero recibió granizo en vez de la lluvia OR in lugar de la lluvia. 17. better! Sra (...
November 26, 2014
Part I looks fine although el almuerzo de ayer might just be el almuerzo ayer? Part II #4 quisieron has a special meaning in the Preterit = they intended, so perhaps querían is better. #7 tuve also has a special meaning in the Preterit = I got/grabbed, etc. so ten&...
November 26, 2014
¡excelente! sin problema Sra I won't even flag this!
November 20, 2014
First sentence is perfect! No, no article with été here. Handy hint: all words ending in -tion are feminine. J'aime visiter des endoits différents pendant (during) les jours de fête. (in = dans, if you prefer) Sra (aka Mme) I'll "flag&...
November 19, 2014
1. sí 2. Después de escribir una carta, la pondría en el buzón. (After a preposition like "después de" the only form of a verb you can use is an infinitive. Because the verb in the question is in the Conditional, your answer should ...
November 14, 2014
You need "clause relater que" = Un sport que la fille n'aime pas c'est nager. (A sport THAT the girl doesn't like is swimming.) The c'est refers back to something prior that you have said. Yes, if you are in 8th Grade, USA. Sra (aka Mme)
November 14, 2014
ma mère Sra (aka Mme)
November 10, 2014
Spanish--1 question
The reason is that the ending of the conjugated verb "tells" the subject. o - yo s - tú vowel (a, e) = él, ella, Usted -mos = nosotros/nosotras -is = vosotros/vosotras -n = ellos, ellas, ustedes Above are the ending signals just for the Present Tense, ...
November 8, 2014
2. hace makes it a run-on sentence. The "ha" you see is half of the Present Perfect = hecho 3. this structure is hace + a period of time to say ...years ago (literally "it makes 20 years that...") The instructions do not say to use each word only once; in ...
November 7, 2014
Without seeing the English of what you are trying to say, I can't always be sure. 1. les magasins = to do or make stores? 2. The accents in préférer go in different directions. Why? Because there are 3 "e" sounds. e = uh / é = a (as first ...
November 6, 2014
Oh yes, I do! Things pile up so quickly and I lose them in the shuffle! Sra
October 31, 2014
Part I: 1. OK 2. best word order for a question is: Interrogative word (if there is one) + verb + subject = ¿Había comido usted algo diferente anoche? 3. pedido instead of ordenado (straightened/ordered) No creo uses the Subjunctive = No creo que haya pedido ...
October 31, 2014
OOPS, Sorry I didn't notice they were negative sentences 2. lire des romans 3. manger de chocolats Sra (aka Mme)
October 31, 2014
OOPS, Sorry I didn't notice they were negative sentences 2. lire de romans 3. manger de chocolats Sra (aka Mme)
October 31, 2014
Only one error here, both in #2 and #3. Don't forget to elide the following: de + le = du, de + les = des, à + le = au à + les = aux. I'll flag this in case you have further questions, so I'll come back later. Sra (aka Mme)
October 31, 2014
When using the Preterit, as well as other words in Spanish , please don't forget the necessary accent marks. If the word has an accent and i t is missing, it is wrong. If you don't know how to do accent marks on the computer, I need to know 2 things: 1) Do you have a ...
October 26, 2014
¡perfecto! Sra
October 23, 2014
If you think this through in Engish, it would be much easier to tell you! This way, I can't think of much! 1. My name is... = Je m'appelle... 2. Let me introduct myself = je me présente 3. I am = Je suis... 4. My first name is = Mon prénom est... 5. I ...
October 20, 2014
The minute you say "I used to..." you are using the Imperfect. I used to roller skate = (yo) patinaba sobre ruedas (one way ) I used to jump rope = (yo) saltaba a la comba I used to ride my tricycle = (yo) montaba mi tricicleta Sra I'll flag this and return later...
October 20, 2014
The difference in French is: photo = la photo photograph = la photographie photography = also la photographie photographer = la photographe For #2 & #3, often you can tell by the context, or exactly how the word is used. Sra (aka Mme) I'll flag this so I can return later ...
October 20, 2014
Olé = well done #4. Often the 2nd negative is after the verb, but it IS flexible! Sra
October 19, 2014
Well done! For #3 se sientan (sentar is to seat someone else but sentarse is to seat oneself!) #4 You could also use the reciprocal "se intercambian" they exchange with each other. How clever of you to catch the y = e>i. Yes. I'll flag this and come back later...
October 19, 2014
To: SraJMcGin
Because this is a Reflexive Verb, it will use être in the passé composé and because the subject is maman (feminine, singular) the past participle will also reflect that gender and number s'est fâchée Sra (aka Mme)
October 19, 2014
French: ms sue or any French expert
Hopefully you saw that I answered that one separately, because it was sent to me separately! Sra (aka Mme)
October 18, 2014
French: ms sue or any French expert
Let's begin with the difference between l'imparfait and le passé composé and how to conjugate each. L'imparfait has these endings for an -er verb: parler = je parlais, tu parlais, il/elle/on parlait, nous parlions, vous parliez, ils/elles parlaient. ...
October 18, 2014
Spanish 3
1. He gives the apple TO HIS BROTHER = TO HIM. This is an indirect object pronoun (whenever either TO, FOR is stated or understood) The Indirect Object Pronouns are: me, te, le, nos, os, les. In this case it is LE. (Le becomes SE ONLY when lo, la, los, las follows it. ...
October 18, 2014
#9 is correct! Sra
October 17, 2014
Part II 9. The verb is not preguntar but esperar = espérenme ***I believe you typed #10 twice, once as #9. You need to retype #9 and it probably IS preguntar! Part III 16. The subject is yo, not tú = diga Everything else looks fine! It is a pleasure to help you, ...
October 17, 2014
Spanish 1
Good Grief! I answered this but it is on a preceding French one that this was attached to. Let me do it over here. Because you began this in the Past Tense (pretérito) I'd change "Cuestan $20" to "Costaron." There is a huge difference between &...
October 16, 2014
Yes, that IS the Subjunctive! :) Sra
October 15, 2014
Hopefully you will get this in time. I didn't see it until this morning (Thursday.) The "si quieren vestirse is only to make the verb "to get dressed/to dress oneself? reflexive. considerar is an -ar verb and you are using the Subjunctive "may consider"...
October 15, 2014
Perhaps "la vestidura" for clothing is better? (feminine) Because the sentence is long, do y ou think "y no debe interrumpir" is a bit clearer Ser is stronger than estar with "es prohibido" or "se prohibe" How about "el ropaje"...
October 15, 2014
Est-ce que vous avez une réponse? Si cet exercice est un devoir, nous ne faisons pas les réponses mais nous aidons avec VOTRE réponse! Sra (aka Mme) (I'll flag this and come back later to see what you write.)
October 15, 2014
Spanish Work Check
2. gender error = la casa 3. some textbooks use the article with Perú = el Perú 8. also with Brasil = al Brasil en el verano The rest looks OK. Sra I'll flag this to come back later in case you have further questions.
October 13, 2014
Spanish Help
With regular formal commands, be sure to use "the opposite vowel" for the formal command. hablar = hable / comer = coma / escribir = escriba 1. You are looking at the tú or informal command, because the formal command is Coma 2. You are now looking at the ...
October 13, 2014
Spanish Check
You are entirely welcome! Sra
October 12, 2014
Spanish Check
1. correct 2. Be sure to study my note at the bottom, with the uses of para and por. In this one is "during a period of time." por la tarde 3. This is "in exchange for something" por $13. 4. When you can substitute "porque = because" it will be ...
October 12, 2014
Part I Sí, por favor, dirá que Anna la gustaría hablar con él. This should be dígale, not dirá, que a Anna le gustaría hablar con él. (you are giving a command to the person on the other end of the telephone) Sr. Costa se...
October 10, 2014
Social Studies
Tahn - kah - DARE The French a is sounded like ah. The spelling "dere" with no accent mark, is pronunced like the American word "dare." Sra (aka Mme)
October 7, 2014
You are entirely welcome! Sra
October 2, 2014
1. OK 2. The verb "gustar" is best learned as meaning "to be pleasing to" because it is usually used 3rd person singular (1 thing is pleasing) and 3rd person plural (2 + things are pleasing to) = The subject is actually "a good grade" would be ...
October 2, 2014
Urgent--Spanish help! :(
First of all, I do not see YOUR answers. It looks like you would like someone else to do your homework? Not fair! Please select the answers YOU believe correct and then I will help you. I will flag this so I can come back later to see your answers. Sra
September 30, 2014
French: TO MS SUE OR ANY FRENCH HELPER
First of all, capitalize a proper name, regardless of the language. Have end punctuation where necessary. French words that require accent marks are incorrect if they are missing those accent marks. You seem unaware of the difference between the Past Tense and the Present ...
September 28, 2014
Part I 1. either cuando (with the Subjunctive) or si (which does not) = cuando se case (whenever she marries) 2 - 4 = OK 5. la familia is singular (a collective unit) = llegue Part II 6. cómelas (drop that extraneous "s") 7. same error = ayúdala 8 & 9...
September 26, 2014
Spanish--check my answers please?
It is always a good idea to number your posts so we refer to the same one. I agree with your answers ***** and then it seems you have not selected an answer. conversations with neighbors & friends = la plaza mayor gaucho games showcase the skills of the gauchos (la Argentina) ...
September 20, 2014
Since I don't know how far you have gotten in studying the Subjunctive, let me know where you are. There are specific reasons to use it. If it IS happening, HAS happened, WILL (for sure) happen, it is the Indicative. If there is doubt, emotion, imposition of will, etc., it...
September 18, 2014
The Subjunctive is the "thinking person's tense." Happy to help! Part I 1. Correct in that it IS the Subjunctive, BUT the subject is "el dependiente" = él = tenga 2. OK 3. Es posible will require the Present Subjunctive & the subject is "la ...
September 18, 2014
You never answered the first post, which I flagged so I could answer any questions you had. I still don't know if the question is asked of YOU about someone else or if it is about you. I need to know if the question asks: 1) Who recommended that dish to him, or her. or 2) ...
September 18, 2014
First of all, you should post the directions. What have you been asked to do? Without knowing that, I'm not sure what to do with this. Also, numbering them is helpful. 1. I assume there is a question, which you are to answer? If so: ¿Quién le recomendó...
September 16, 2014
Urgent: Spanish, Check Answers Please
"de nada" = You are entirely welcome! Sra
September 9, 2014
Urgent: Spanish, Check Answers Please
6. Your answer is incorrect and here is why. The subject is plural = Claudia y María = Ellas, or 3rd person plural. The verb must also be 3rd person plural. Here are the forms of ser: lst person singular = soy (yo) 2nd person singular = eres (tú) 3rd person ...
September 9, 2014
spanish please check!
It's best to always begin with the instructions of what you are to do. If you are studying the Preterit, learn the endings for regular verbs and then the irregular verbs. 1. The verb "ir" is irregular in the Preterit and looks exactly like the verb ser except the...
August 25, 2014
Spanish use of Imperfect Verbs
OK. Thanks and ¡Buena Suerte! Sra
July 26, 2014
Spanish use of Imperfect Verbs
1. But before I finish = Pero, antes de terminar... 2. You are using the vosotros,-as form? Many textbooks do not include it. If you are, here is the choice for "And I would like to give you some advice" = Y me gustaría daros algunos consejos... Párrafo...
July 26, 2014
Spanish use of Imperfect Verbs
First word is spelled "Hola,.....familias; me llamo Nick y estoy en.......de la Argentina En la mañana is better grammar with "Por la mañana..."íbamos (acento)..de las partes para las políticas = for female politicians OR para la pol&...
July 26, 2014
Spanish 3
This is a "si" clause, when "si" means "if/whether" and the English translation could be variable. If I knew (were to know, would knowo) a film star, I would ask for his/her autograph and I would put it on the wall of my room. d. the "si"...
July 25, 2014
Spanish 3
In this one (with the "si clause") the main clause (Independent) comes first and the "si clause" (in the Dependent clause) comes second. Here's the English for Mihir: We would have finished all the household chores if our mother had not returned/would ...
July 25, 2014
I'm glad! I also have lists of verbs imposition of will, emotion, doubt, etc. and any examples you might want. Sra
July 17, 2014
If you need the list of verbs for #1, etc., just ask. Sra
July 17, 2014
#3. Here is another example of "Imposition of Will" which is one of the main reasons to have the Subjunctive. Have you had a good review of WHY the Subjunctive exists? It is rapidly dying out in English but still alive and well in Spanish and French. NOUN CLAUSES: ...
July 17, 2014
Spanish help
2. I suggest that YOU prepare = A / Sugiero requires the Subjunctive (Imposition of Will) 3. restaurante is misspelled. The rest is correct (compare #3 with #2) 4. salsa is not made of cheese but stuffing is = B 6. Es necesario is also Imposition of Will and requires the ...
July 16, 2014
3. A. a double room / B has a spelling error 4. D. is the correct answer because "ve" is other the tú command for "go" or "see" The rest are correct. Again, I'll flag this so I can come back later to see if you have any other questions...
July 14, 2014
Spanish help
Good. I'm glad! Sra
July 13, 2014
Spanish help
P.S. I flagged this, so in case you have any questions, I'll come back later to check. Sra
July 13, 2014
Spanish help
2. no habrá = there will not be 3. A is correct but be sure you spell it correctly = será 4. The question asks "what WAS" (past tense) so the answer must also be in the past tense = A. 5. You selected the correct answer = will be able. B said he will (...
July 13, 2014
There is nothing wrong with A. B could be incorrect if you are NOT referring to 2 people.' Sra
July 7, 2014
First of all, it would be very helpful if you would number the items = 1., 2., 3., etc. The first one has 2 errors, both B & C. B. ¿Dónde está Maria? (location) C. Ella es de México. (origin) The rest are OK in the first set. In the 2nd set, the ...
July 7, 2014
Thank you for providing the English of what you wish to say in Spanish. There are, however, some grammar problems there! 1. double negative = not doing anything...2. How is it possible = implies a question so one would expect a question mark at the end = ? 3. The number of ...
July 6, 2014
literally with the Present Perfect Tense Decide if you want familiar (tú/vosotros/vosotras), singular or plural, or formal (usted/ustedes) I'll give you the tú form: ¿Has estado estudiando tu manual de conductor? It would be more likely to use the ...
July 1, 2014
11. More likely to be: Ellos tienen tantos hijos como nosotros. (because we are comparing "ellos" to "nosotros." 13. Learn the verb "gustar" as the verb "to be pleasing" and you might have less difficulty, because it is usually used in ...
June 21, 2014
1. used to sing OR was singing OR even sang is translated by the Imperfect Tense. = El rey que cantaba está tocando el tambor. There are two verbs for "to play." "To play a game" is "jugar" BUT "to play an instrument" is "...
June 20, 2014
Spanish Preterite/Imperfect Sentences
This is a difficult one to explain. I can onl give you examples. If you are standing in one place and what to say "something is seen FROM that spot where you are standing = desde. "From the beach I can see the hotel." "the view is wonderful from the church...
June 12, 2014
Spanish Preterite/Imperfect Sentences
3. although both "desde" and "de" are prepositions, there is a difference in meaning. "desde" means since, from, after while "de" means of, from, about. In the expression "the view was great FROM the hotel - I would then use "...
June 12, 2014
Spanish Grammar
You are entirely welcome! Any time! Sra
June 11, 2014
Spanish Grammar
A bit better with "por todo el mundo" and "Me construiría" means you don't really need "para mí" = Me construiría unas casas por toeo el mundo" = the rest seems a bit redundant. If you are building houses for yourself...
June 11, 2014
Spanish Grammar
1. Perhaps, instead of "durante" you might like "por" = for 7. Non modified nouns often use the article (el, la, los, las) = me gusta el tocino arar el desayuno) 8. In Spanish, always verify the gender (masculine/feminine) and number (singular/plural) of ...
June 11, 2014
Yes! You have the idea. We don't worry about gender in English, but you do in other languages. Good job! Sra
June 4, 2014
Because the questions are in English, they should be very clear to you. 1. Always check number (singular/plural) and gender (masculine/feminine) of all adjectives. "Esta muchacha" is feminine and singular BUT the adjective "bonito" is singular and masculine...
June 4, 2014
8. I prefer it with the accent mark, but you may see it without, because, according to the rule of pronunciation, if the word ends in a vowel (a, e, i, o, u) the stress is on the next-to-last syllable. habla / hable / dame / deme ¡perfecto! Sra
May 29, 2014
They are not the only 2 tenses used in the Past. There is the Conditional (would) = the past of the future! There are also 2 forms of the Past Subjunctive (-ra/-se) and the Perfect Tenses (have/has + past participle, would have + past participle, + the Past Perfect Subjunctive...
May 23, 2014
First of all, you need to correct the question. The Preterit is used when the event has a beginning and an end. But "estuve" is the "yo" form and not the "it = el fin de la semana" which is "estuvo" in this case. However, the Imperfect ...
May 23, 2014
Yes, initials would be good to know who is speaking to whom. Don't forget the accent marks, for if you are turning this on, all the words requiring accent marks will be incorrect. A ¡Hola! (2 exclamation marks) ¿Qué pasa? (2 question marks plus accent) ...
May 23, 2014
¿En vez de "aquí' considera "allí"? (línea 4) En la frase penúltima es necesario tener "que" o: La característica más singular es que el mihrab se enfrenta...... o La característica más...
May 21, 2014
Did you do this backwards? The numbers come first and then th e corresponding letter! However, it looks like you got them all! Here's what I have: 1. d 2. g 3. a 4. e 5. f 6. b 7. c Sra
May 21, 2014
Please allow me to explain the difference between "help" and "do." You are NOT asking for help but asking me to DO the work for you. That I will NOT do. First of all, get a good dictionary. Look up each word trying to find one of the choices below (a, b, ...
May 21, 2014
Spanish 2
1. You can also use the Imperfect = Compraba (unos/los) libros para los estudiantes. IF you must use a direct ot indirect object pronoun, and those were the instructions = Les compraba libros para (a) los estudiantes. 2. correct but with pronoun = El desayuno, lo hemos comido...
May 20, 2014
Spanish 2
#1. There is a difference between "el cine" = the movie theater and "la película" = the movie/film. #2. Again, you might use the imperfect = Ibamos al parque. #3. Hicieron la tarea = just another way. Sra
May 19, 2014
Spanish 2
Since you do not give more explicit directions, merely saying "Rewrite in Spanish" #1 is one way. Another way is with the imperfect which translates I used to eat, I was eating = comía... #2. This is merely the masculine singular form of the adjective OR the ...
May 19, 2014
Spanish (Please help me)
The answers are on the first one I so, so please go there. Sra
May 16, 2014
Yes, of course you could! Sra
May 16, 2014
First of all you need to learn how to do tú (or familiar) commands, both affirmative and negative. Regular verbs look like the 3rd person singular in the affirmative (hablar = habla)(comer = come) (vivir = vive) BUT in the negative they use the Subjunctive, which is ...
May 16, 2014
13. Sorry, I misread that and didn't realize that "estudiar" was there twice. You were correct then, as this is the entire sentence: Nosotros tenemos que estudiar español cada día pero anoche estudiamos biología también. My error! ...
May 15, 2014
1 - 6 = perfect 7 = por, because it means "in a period of time" 8 = perfect 9 = por = "for the sake of" / NOTE: often, in place of "para" you can substitute "a" and in place of "por" you can substitute "porque" 10 = ...
May 15, 2014
los toros son espantosos y de mal genio. How to use a dictionary: 1. take the Engtlish word and look up all the Spanish possibilities and write those down. 2. mean = medio, humilde, obscuro, andrajoso, raído, mezquino, tacaño, inferior, pobre, vil, ruin, innoble...
May 8, 2014
Paragraph 2 = Son cinco compañeras might better be Eran? #3 los toros me dan miedo. (son de miedo y media? = not sure what you were saying there) Sra Again, I'll flag this and come back later, in case y ou have any questions.
May 8, 2014
Spanish Proofreading
That looks so much better! The first should have been "teníamos mucha diversión" but I like what you did with it better! Sra
May 4, 2014
Spanish Proofreading
la nieve = feminine/singular so the adjective must be also = rara esperé a mis padres = personal a" only, no clause relater "que" La Señora Andrews = when speaking about her, the honorific article is used (for politeness) por = on behalf of her ...
May 4, 2014
retourner = to go back return, because "rendre" is to return an unwanted item! tous les sites que (not qui) and what is "sur le mien" supposed to mean (on mine) D'abord, irais (je is the subject, not il/elle)...to learn? = pour apprendre plus = to learn...
May 4, 2014
Sra is the abbreviation for Señora! I am not a "sir!: Yes, of course, I'll help you. I have flagged your 3 questions so I can come back to see if you have any further questions. Sra
May 3, 2014


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APARTMENTS FOR RENT: MIAMI -2/2 - 1st fl, spacious, W/D walking closet,2 park,close to FIU $1,500 - 305-389-4741More info Email a friend
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Browsing by Subject "Impressions"
Now showing items 1-1 of 1
A House For Stories (Virginia Tech, 2013-02-18)A house can be considered a place made from the stories that make it. In order to design the house I recalled places I experienced during my childhood. Digging back through the memories of those places I became intrigued ...