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Plants are sessile and therefore have developed mechanisms to adapt to their environment, including the soil mineral nutrient composition. Ionomics is a developing functional genomic strategy designed to rapidly identify the genes and gene networks involved in regulating how plants acquire and accumulate these mineral nutrients from the soil. Here, we report on the coupling of high-throughput elemental profiling of shoot tissue from various Arabidopsis accessions with DNA microarray-based bulk segregant analysis and reverse genetics, for the rapid identification of genes from wild populations of Arabidopsis that are involved in regulating how plants acquire and accumulate Na+ from the soil. Elemental profiling of shoot tissue from 12 different Arabidopsis accessions revealed that two coastal populations of Arabidopsis collected from Tossa del Mar, Spain, and Tsu, Japan (Ts-1 and Tsu-1, respectively), accumulate higher shoot levels of Na+ than do Col-0 and other accessions. We identify AtHKT1, known to encode a Na+ transporter, as being the causal locus driving elevated shoot Na+ in both Ts-1 and Tsu-1. Furthermore, we establish that a deletion in a tandem repeat sequence approximately 5 kb upstream of AtHKT1 is responsible for the reduced root expression of AtHKT1 observed in these accessions. Reciprocal grafting experiments establish that this loss of AtHKT1 expression in roots is responsible for elevated shoot Na+. Interestingly, and in contrast to the hkt1–1 null mutant, under NaCl stress conditions, this novel AtHKT1 allele not only does not confer NaCl sensitivity but also cosegregates with elevated NaCl tolerance. We also present all our elemental profiling data in a new open access ionomics database, the Purdue Ionomics Information Management System (PiiMS; http://www.purdue.edu/dp/ionomics). Using DNA microarray-based genotyping has allowed us to rapidly identify AtHKT1 as the casual locus driving the natural variation in shoot Na+ accumulation we observed in Ts-1 and Tsu-1. Such an approach overcomes the limitations imposed by a lack of established genetic markers in most Arabidopsis accessions and opens up a vast and tractable source of natural variation for the identification of gene function not only in ionomics but also in many other biological processes.
Unlike most animals, plants are sessile and cannot leave a poor-quality environment after germinating. They therefore need to tolerate the particular conditions they encounter to survive. This makes plants an ideal system for the study of adaptive variation, and this is particularly true of Arabidopsis thaliana (Arabidopsis), which shows substantial natural variation and for which numerous genetic tools exist. Using a combination of analytical chemistry, genetics, and genomics, the authors were able to identify the specific genetic alteration that drive the natural variation in shoot sodium (Na+) accumulation capacity observed in Arabidopsis populations from coastal regions of Spain and Japan (Tossa del Mar and Tsu, respectively). They observed that a deletion in the DNA responsible for regulating the expression of HKT1, a gene known to encode for a Na+ transporter, causes reduced expression of AtHKT1 in roots of both the Spanish and Japanese populations. Such altered expression results in the elevated shoot Na+ observed in these two populations. Interestingly, this novel version of the HKT1 genes is also associated genetically with the enhanced NaCl resistance they observe in the Japanese population.
Plants are sessile and therefore have developed mechanisms to adapt to their environment, including the soil mineral nutrient composition. High-throughput elemental profiling of Arabidopsis thaliana (Arabidopsis) has been used in an effort to identify the genes and gene networks involved in regulating how plants acquire and accumulate mineral nutrients and trace elements from the soil [1]. In 2003, Lahner et al. [1], in a screening of 6,000 fast-neutron–mutagenized Arabidopsis plants grown under unstressed conditions, identified 51 mutants with altered shoot elemental profiles, and they estimated that about 2% to 4% of the Arabidopsis genome is involved in regulating the elemental composition or "ionome" of Arabidopsis (for review, see [2]), including accumulation of macronutrients, micronutrients, and nonessential elements such as Na+. Recently, one of these ionomic mutants was shown to harbor a deletion in AtHKT1 that is responsible for the elevated shoot Na+ phenotype of this mutant [3].
As an alternative to induced mutations (fast-neutron, ethylmethane sulfonate, etc.), the large reservoir of natural variation that exists in Arabidopsis is also a potentially powerful resource for the investigation of ionomic gene function [4–6]. Such natural variation has the advantage over induced mutations in that uncovering the adaptive significance of such variation provides tools for the integration of gene function in the context of whole plant physiology. However, this genetic resource is still underexploited, mainly because natural phenotypic variation is usually the result of genotypic variation at multiple loci. Even when dealing with monogenic traits, it is a major challenge to identify a particular gene controlling a phenotype of interest. Currently, fewer than ten genes have been identified in Arabidopsis using the natural variation approach [7,8], whereas variation in multiple traits such as floral and meristem development, resistance, and defense against pathogens as well as metabolic enzymes ([9] and references therein) have been documented. Genetic differences between local populations are presumably associated with adaptation to the prevailing environmental conditions, although well-established examples of this in Arabidopsis are limited. Such investigations are impeded by the fact that very little information exists about the environmental conditions and habitat of the Arabidopsis accessions that have been collected and are curated at the Arabidopsis Biological Resource Center. The term "accession" is used to refer to natural genetic variants of Arabidopsis to avoid the term "ecotype," which would allude to a variant genetically adapted to a particular habitat [10,11].
Here, we have coupled high-throughput elemental profiling of shoot tissue [1] from various Arabidopsis accessions with DNA microarray-based bulk segregant analysis (BSA) [12,13] and reverse genetics, for the rapid identification of genes involved in regulating how plants acquire and accumulate Na+ from the soil. Such an approach using DNA microarray-based genotyping overcomes the time constraints imposed on using natural variation by the rapid identification of genetic markers, and opens up a vast and tractable source of natural variation for the identification of gene function not only in ionomics but also in many other biological processes. In a system with multiple quantitative trait loci, an alternative method, eXtreme Array Mapping, was developed to identify major quantitative trait loci using pools of lines with extreme phenotypes [14].
Using this approach, we have identified a novel AtHKT1 allele from two different Arabidopsis accessions collected from Tossa del Mar (Ts-1) and Tsu (Tsu-1), sites located on the coastal regions of Spain and Japan, respectively. This novel AtHKT1 allele, in both Ts-1 and Tsu-1, is responsible for the higher shoot Na+ accumulation observed in these accessions compared with the reference accession Col-0, when grown under background NaCl levels. Previously, expression of AtHKT1 has been shown to be localized in the vascular tissues throughout the plant, with expression being highest in the roots [15–17]. The hkt1 null mutation results in an alteration of the Na+ distribution in the plant, with higher levels of Na+ accumulation in shoots and lower accumulation in roots compared to wild-type plants. Loss of AtHKT1 expression in the null hkt1 mutant also confers sensitivity to high levels of NaCl [17–20]. All these studies are consistent with the current model that HKT1 functions to tightly regulate Na+ accumulation in shoots by unloading Na+ from the xylem in both roots and shoots and recycling Na+ to the roots via the phloem.
Results reported here confirm that this new naturally evolved AtHKT1 allele is responsible for the elevated shoot Na+ accumulation we observe in Ts-1 and Tsu-1, via the specific downregulation of AtHKT1 expression in roots. Reciprocal grafting also confirms that the primary site for HKT1 function in regulating shoot Na+ is in the root, with HKT1 playing little role in the shoot. Further, we suggest a deletion in the promoter region as the factor responsible for driving the differential expression of AtHKT1 in both Ts-1 and Tsu-1. Interestingly, and contrarily to the loss-of-function mutants hkt1, this novel allele appears to also be associated with enhanced NaCl tolerance.
Elemental profiling of shoot tissue collected from 12 different Arabidopsis accessions, including Col-0, Cvi-0, Est-1, Kas-2, Mrk-0, Mt-0, Se-0, Ts-1, Van-0, Nd-1, Tsu-1, and Ler-2, revealed that Ts-1 and Tsu-1 (Figure 1A) accumulate higher levels of Na+ than Col-0 and all other accessions tested when propagated in soil with low levels of NaCl under controlled growth conditions (Figure 1B). Interestingly, the Ts-1 and Tsu-1 accessions were collected from the coastal regions of Spain (Tossa del Mar) and Japan (Tsu), respectively. Besides being annotated as having different geographical origins, Ts-1 and Tsu-1 have different physical appearances (Figure 1A). Moreover, based on a genome-scale DNA polymorphism analysis involving 96 Arabidopsis accessions, these two accessions appeared to be genetically distinct [21]. The Ts-1 and Tsu-1 accessions can also be distinguished based on the fact that Ts-1 accumulates elevated concentrations of cobalt (+55%) in shoot tissue compared to Col-0, while Tsu-1 has lower levels of Co. A complete set of the elemental profiling data summarized here can be obtained at the open access Purdue Ionomics Information Management System (PiiMS) at http://www.purdue.edu/dp/ionomics.
To determine the feasibility of using natural variation to identify individual loci regulating ionomic variation, we attempted to map the locus responsible for elevated Na+ in Ts-1 and Tsu-1. Both accessions were crossed to Col-0, and the shoot concentrations of Na+ were measured in the F1 hybrid plants (Figure 1C). The concentration of Na+ was not significantly different from Col-0, suggesting that the loci controlling Na+ accumulation in Ts-1 and Tsu-1 are recessive in both cases. Analysis of the segregating F2 plants from either cross revealed a pattern consistent with 3:1 segregation ratio of Col-0 × Ts-1 (Figure 2A) and Col-0 × Tsu-1 (Figure 2B) for shoot Na+ concentrations. Therefore, we conclude that the high Na+ phenotype observed in both Ts-1 and Tsu-1 segregates as a monogenic recessive trait. We further used the complementation test to demonstrate that the loci driving elevated shoot Na+ in Ts-1 and Tsu-1 are allelic to each other, showing that the F1 plants from Tsu-1 × Ts-1 have shoot Na+ levels that are similar to each parent (Figure 1D). Based on this evidence, DNA microarray-based BSA [12,13] was used to map the locus responsible for elevated Na+ in both Ts-1 and Tsu-1. From the Ts-1 × Col-0 F2 population represented in Figure 2A, two pools of DNA were prepared, one containing genomic DNA from 30 plants with Na+ levels similar to Col-0, constituting the "control" pool, and another with DNA from 30 plants with the highest Na+ levels, similar to those of the Ts-1 parent, constituting the "high Na+" pool. Single feature polymorphisms (SFPs) were identified for Col-0 and Ts-1 by comparing the hybridization of genomic DNA isolated from the parental Col-0 and Ts-1 accessions to the Affymetrix Arabidopsis ATH1 microarrays. The frequency of these SFPs in the DNA from the pool of F2 progeny plants with high Na+ levels, and in the pool with control Na+ levels, was scored by hybridization of genomic DNA to ATH1 microarrays. The DNA microarray-based BSA revealed a strong peak and therefore enrichment in Ts-1 SFPs linked to the locus causing elevated shoot Na+ in Ts-1 at 6.6 Mbp on Chromosome IV (Figure 2C). Similarly, a strong peak at 6.4 Mbp in Chromosome IV was obtained when the hybridization was performed using DNA pools from the Tsu-1 × Col-0 F2 population (Figure 2C). Approximately 513 genes (corresponding to 23 BAC clones) are contained within the confidence interval of 5.525 to 7.475 Mbp on Chromosome IV. The confidence interval was derived using previously published simulations [12]. One of the genes in BAC clone F24G24 (at 6.39 Mb), At4g10310, named AtHKT1, stands out as a very strong candidate because it has been previously identified as a Na+ transporter [15], which when disrupted causes elevated shoot Na+ [3,17,18]. The shoot Na+ concentrations in the hkt1–1 null mutant are significantly higher than Col-0 in leaves at all stages of development, with Na+ accumulation generally increasing with leaf age (Figure S1A). When grown under the same conditions, the Ts-1 and Tsu-1 accessions also show increased Na+ accumulation compared to Col-0 in leaves at all stages of development. However, Na+ accumulation in Ts-1 and Tsu-1 is lower than that observed for hkt1–1 (Figure S1A).
To establish if a different allele of AtHKT1 is present in Ts-1 and Tsu-1, the nucleotide sequence was determined for the AtHKT1 locus in both accessions. The sequenced region comprises 5,456 bp upstream (363 bp downstream of the previous gene At4g10300) of AtHKT1 translational start codon that includes the promoter to 386 bp downstream of the AtHKT1 stop codon. Several polymorphisms were identified both in the coding region and in the upstream and downstream regions of the AtHKT1 gene. In the coding region, no nonsense mutation was recognized and 19 single nucleotide polymorphisms (SNPs) were identified in both Ts-1 and Tsu-1 (Figure S2A), seven of which result in the amino acid changes R3I, K10N, V66L, C134Y, E385G, V453L, and F477L. None of the amino acid changes are located in the three relatively highly conserved regions identified by Uozumi et al. [15], and none of them affect the first serine residue at position 68 of P-loop A that has been implicated in the Na+ specificity of the transporter [22]. One of the amino acid changes, valine to leucine in position 453 (V453L), was also found in the Ler AtHKT1 allele published by Uozumi et al. [15].
In the upstream region, which includes the promoter of AtHKT1, four major polymorphisms were identified between Ts-1, Tsu-1, and Col-0 (Figure S2B). The most upstream polymorphism affects two repeated units of 681 and 673 bp, linked by a 34-bp sequence (Figure S2B). These two repeated units are localized 5,320 to 3,933 bp upstream of the AtHKT1 start codon and are 97% identical to each other in Col-0. In Ts-1 and Tsu-1, we identified a 725- and 687-bp deletion, respectively, which removes the end of the first of these repeated units, the beginning of the second unit, and the 34-bp junction sequence. Consequently, only one of the repeated units is present in Ts-1 and Tsu-1. The second major polymorphism identified in this region, about 3,100 bp upstream of the start codon, is an insertion of 13 bp in the sequence of Ts-1 as well as Tsu-1, resulting in the duplication of a 10-bp sequence (AATGTGTTAT) in Ts-1 and Tsu-1 that is present only once in Col-0. Similarly, 1,210 bp upstream of the start codon, a 14-bp insertion results in a duplicated 10-bp sequence (TCATTGCAAA) in Ts-1 and Tsu-1. The last major polymorphism in the promoter region is localized 148 to 98 bp upstream of the translational start codon. Compared to Col-0, these sequence polymorphisms found in both Ts-1 and Tsu-1 abolish the first of the two putative CAAT boxes suggested by Uozumi et al. [15].
Polymorphisms have also been found in the two introns of AtHKT1, mainly SNPs. We have been unable to sequence the last 157 bp of the second intron of the Ts-1 and Tsu-1 alleles, which according to the published Col-0 sequence is enriched in [TA] repeats. Finally, major differences in the sequence between Col-0 and Ts-1 and Tsu-1 occur in the sequence downstream of the stop codon, including the 3′-untranslated region. Several SNPs, as well as deletions in Ts-1 and Tsu-1, result in a sequence of 246 and 140 bp shorter than the 386-bp sequence found in Col-0.
Because the Ts-1 and Tsu-1 HKT1 alleles are recessive (Figure 1C) and the hkt1–1 null mutant, like Ts-1 and Tsu-1, shows elevated shoot Na+ accumulation (Figure S1), we used the deficiency complementation test to establish that AtHKT1 is the gene responsible for elevated Na+ in Ts-1 and Tsu-1 [23]. Both accessions, Ts-1 and Tsu-1, were crossed to the hkt1–1 T-DNA insertional mutant in the Col-0 gl1 genetic background [19]. The hkt1 null mutant accumulates higher levels of Na+ in the shoot and lower levels in the root than the wild-type, implicating HKT1 in the regulation of Na+ distribution throughout the plant [17–19]. F1 plants from the cross of hkt1–1 to Col-0 present Na+ levels similar to the those of the wild-type, as expected from a recessive mutation (Figure 3). However, F1 hybrid plants from the crosses of hkt1–1 (Col-0 gl1) with Ts-1 or Tsu-1 accumulate levels of Na+ that are intermediate between each parent and significantly higher than Col-0 (Figure 3), indicating that the Ts-1 and Tsu-1 HKT1 allele is deficient in complementing the hkt1–1 null mutation. This helps establish that elevated Na+ in Ts-1 and Tsu-1 is due to alterations in the AtHKT1 gene. However, the lower levels of Na+ in the F1 plants compared with hkt1–1 suggest that the Ts-1 and Tsu-1 alleles of AtHKT1 are not nulls but rather weak alleles, although a role of the genetic background cannot be excluded (Figure 3).
Several polymorphisms in the promoter region suggested that HKT1 in Ts-1 and Tsu-1 might have a different expression level or expression pattern compared to Col-0, which could be responsible for the high Na+ phenotype in Ts-1 and Tsu-1. Plants displaying elevated shoot Na+ accumulation, compared to Col-0, were used to determine expression levels of AtHKT1 in shoot and root tissue, using quantitative real-time PCR. As previously described [15–17], expression of AtHKT1 in Col-0 is significantly higher in roots compared to shoots (Figure 4). In the shoot, no significant differences in expression levels were found between Col-0 and Ts-1 or Tsu-1. In the root, however, the level of AtHKT1 expression in Ts-1 and Tsu-1 is approximately 10-fold lower than in Col-0 (Figure 4).
An Arabidopsis mutant (Line 1425), in the Col-0 gl1 sos3–1 background, previously identified as a weak suppressor of sos3–1 Na+ sensitivity (A. Rus and P. M. Hasegawa, unpublished data), was found to contain a T-DNA insertion within the tandem repeat region localized 5,320 to 3,933 bp upstream of the AtHKT1 start codon (Figure 5A), where Ts-1 and Tsu-1 also have a 725- and 687-bp deletion, respectively (Figure S2B). We observed that in Line 1425, expression of AtHKT1 in both roots and shoots is drastically reduced to levels comparable to hkt1–1 (Figure 5B). Furthermore, disruption of this tandem repeat, and elimination of AtHKT1 expression in Line 1425, leads to elevated shoot Na+ compared to both Col-0 and sos3–1 (Figure 5C), and this phenocopies the elevated shoot Na+ levels observed in both hkt1–1 and the sos3–1 hkt1–1 double mutant (Figure 5C). Such evidence strongly implicates the approximately 700-bp deletion in the tandem repeat upstream of AtHKT1 in both Ts-1 and Tsu-1 as being involved in the reduced root expression of AtHKT1 in these accessions.
The elevated shoot Na+ of Ts-1 and Tsu-1 that maps to the AtHKT1 locus is associated with the loss of AtHKT1 expression in roots. To address the contribution of HKT1 expression in the root to the control of Na+ accumulation in the shoot, we performed reciprocal grafting experiments (Figure 6). Shoots from hkt1–1 grafted onto Col-0 roots show no significant difference in shoot Na+ compared to Col-0 self-grafted controls, while Col-0 shoots grafted onto hkt1–1 roots accumulated Na+ levels similar to those observed in self-grafted hkt1–1 plants (Figure 6). This is further evidence that AtHKT1 expression in the root is the primary site for HKT1′s role in regulating shoot Na+.
AtHKT1 has been implicated in Arabidopsis tolerance to NaCl stress. The hkt1 null mutant is sensitive to elevated NaCl as a result of Na+ overaccumulation in the shoot [Figure 7A; 17–19]. Under unstressed conditions, we have established that the weaker AtHKT1 allele in Ts-1 and Tsu-1 is responsible for the elevated Na+ content in the shoot of these two accessions, similar to that observed in the hkt1–1 null mutant. Under high NaCl concentrations (50 and 100 mM NaCl), both Ts-1 and Tsu-1 have higher levels of Na+ in the shoot compared to Col-0, in all leaves at different stages of development, including the youngest ones not fully developed and localized close to the meristem (stage 1; Figure S1B and S1C). However, this elevated shoot Na does not cause increased NaCl sensitivity in Tsu-1 (Figure 7A). Given the connection between AtHKT1 expression and NaCl sensitivity, we decided to evaluate the involvement of this novel allele of AtHKT1 in NaCl tolerance by screening F2 plants from a cross between Tsu-1 × Col-0 for their capacity to survive high levels of NaCl (Figure 7). The genotyping of the F2 population from Tsu-1 × Col-0 identified 25 plants homozygous for Tsu-1-HKT1 and 50 plants heterozygous and 20 plants homozygous for the Col-0-HKT1 allele, following the expected 1:2:1 segregation ratio. The plants homozygous or heterozygous for the Tsu-1-HKT1 allele were able to survive longer in the presence of 100 mM NaCl compared to plants that were homozygous for the Col-0-HKT1 allele (Figure 7). Therefore, although this new AtHKT1 allele contributes to an increased Na+ accumulation in the shoot, it does not confer NaCl sensitivity but, on the contrary, appears to co-segregate with enhanced NaCl tolerance. Although currently we cannot eliminate the possibility that a second unknown gene is conferring the enhanced NaCl tolerance observed in Tsu-1, and due to a small genetic distance between AtHKT1 and this second gene they segregate together in the F2 population studied. Further transgenic studies are required to discern this.
We have used natural variation in Arabidopsis to successfully identify a new AtHKT1 allele responsible for elevated Na+ accumulation in the shoots, compared to Col-0. We propose the reduced expression of AtHKT1 in roots as the cause of this enhanced Na+ accumulation and provide evidence which points to a deletion in a tandem repeat upstream of AtHKT1 as responsible for this differential regulation of AtHKT1 expression. Identification of AtHKT1 as a candidate gene was possible based on the mapping position obtained from our DNA microarray-based BSA analysis, combined with the known phenotype of hkt1. However, the difficulty arises in providing significant proof that AtHKT1 is the causal gene driving elevated shoot Na+. To achieve this, we have combined several lines of evidence as suggested by Weigel and Nordborg [23]. First, DNA polymorphisms allow us to distinguish between the AtHKT1 allele in Col-0 and the AtHKT1 allele found in both Ts-1 and Tsu-1. Second, the deficiency complementation test indicates that the new AtHKT1 allele found in Ts-1 and Tsu-1 is responsible for the Na+ accumulation in the shoot. Third, AtHKT1 shows a different expression pattern in Ts-1 and Tsu-1 compared to Col-0. Fourth, the characterization of this new AtHKT1 allele was performed in two independent genetic backgrounds, Ts-1 and Tsu-1, with the same results.
High-throughput elemental profiling of Arabidopsis mutagenized lines has proved to be very successful in identifying mutants with altered ionomes, some of which are caused by variation at a single locus [1,3]. As presented here, we have also found, using high-throughput elemental profiling coupled with genetics and DNA microarray-based mapping techniques, that the higher Na+ levels found in the shoots of Ts-1 and Tsu-1, two different Arabidopsis accessions, are due to natural variation at the AtHKT1 locus. AtHKT1 has been shown to transport Na+ and, based on studies involving the hkt1 null mutants, has been implicated in Na+ distribution throughout the plant [15,17,18].
The concentrations of Na+ found in the shoots of Ts-1 and Tsu-1 are intermediate between the hkt1–1 null mutant and Col-0 (Figures 3 and S1). This, coupled with the moderate complementation of the high Na+ phenotype of hkt1–1 observed in the F1 plants from an hkt1–1 × Ts-1 or Tsu-1 cross, suggests that the AtHKT1 allele in these accessions is likely not a null, but rather a weak, allele (Figure 3), although a genetic background effect cannot be excluded. Several amino acid changes occur in HKT1 from Ts-1 and Tsu-1; however, no nonsense mutation or frameshifts were found in the coding region (Figure S2A). Interestingly, deletions upstream of AtHKT1 are present in both Ts-1 and Tsu-1, and analysis of AtHKT1 expression revealed it to be dramatically reduced in roots of both Ts-1 and Tsu-1 but unaltered in shoots, compared to Col-0 (Figure 4). Such changes in expression levels are likely to be responsible for the accumulation of Na+ in the shoots of Ts-1 and Tsu-1.
Previously, a suppressor of the NaCl sensitivity of sos3–1 (Col-0 gl1) called Line 1425 (A. Rus and P. M. Hasegawa, unpublished data) was identified as having a T-DNA insertion within the tandem repeat region upstream of AtHKT1 (Figure 5A), the same region that contains an approximately 700-bp deletion in both Ts-1 and Tsu-1 (Figure S2B). The reduction in AtHKT1 expression in Line 1425, which is associated with elevated shoot Na+ (Figure 5C), supports our hypothesis that the deletion in the tandem repeat region between 5,320 to 3,933 bp upstream of AtHKT1 is responsible for both the reduced expression of AtHKT1 in roots and elevated shoot Na+ observed in both Ts-1 and Tsu-1. Based on our sequence analysis and previous promoter-GUS reporter gene analysis [20], this tandem repeat region is well upstream of the promoter of AtHKT1. However, further analysis of that region in the Arabidopsis mpss database ([24]; http://mpss.dbi.udel.edu/at) revealed the presence of a dense cluster of small RNAs from the unannotated tandem repeat upstream of AtHKT1. We hypothesize that because this tandem repeat region appears to be involved in positively regulating expression of AtHKT1, siRNAs or miRNAs produced from this region may be involved in interfering with a system that negatively regulates AtHKT1 expression. However, confirmation of this model requires further experimentation.
The availability of the natural variant of AtHKT1 in Ts-1 and Tsu-1, in which AtHKT1 expression is altered specifically in roots, supports a model for HKT1 function in which the transporter plays a role in controlling Na+ accumulation in shoots by regulating, at the root level, the transport of Na+ in the xylem transpiration stream to the shoot. We propose that lower expression of AtHKT1 in the roots results in reduced Na+ retrieval from the root xylem, leading to the elevated Na+ observed in Ts-1 and Tsu-1 shoots. Such a model is strongly supported by our grafting experiments which clearly demonstrate that activity of HKT1 in the roots is the major site of HKT1 function in regulating Na+ accumulation in shoots (Figure 6). These grafting experiments also demonstrate that the specific loss of HKT1 function in shoots has no significant effect on shoot accumulation of Na+ (Figure 6). This evidence refines the existing model which proposes that HKT1 also plays a significant role in retrieving Na+ from shoots and exporting Na+ from the shoot to root via the phloem [17,20], for which we find no evidence.
The Arabidopsis Na+ transporter HKT1 plays an important role in the shoot Na+ avoidance strategy of Arabidopsis, and because the new AtHKT1 allele reported here results in higher accumulation of Na+ in shoots (Figures 1, ,3,3, and S1), we would expect plants containing this allele of AtHKT1 to be more sensitive to NaCl stress, as compared to Col-0. However, we observed this not to be the case. A comparison of the Col-0 and Tsu-1 ability to survive treatment with Na+ clearly shows that Tsu-1 survives longer than Col-0 (Figure 7). Furthermore, F2 plants from a cross between Col-1 × Tsu-1, either homozygous or heterozygous for the AtHKT1 allele from Tsu-1, were more NaCl tolerant than plants homozygous for the Col-0 AtHKT1 allele (Figure 7D). Such cosegregation of the Tsu-1 AtHKT1 allele with increased NaCl tolerance is consistent with this new allele of AtHKT1 being involved in the elevated tolerate to NaCl observed in Tsu-1. This evidence also confirms that in Arabidopsis, reduced shoot accumulation of Na+ is not necessary for enhanced salinity tolerance [25].
Both the Ts-1 and Tsu-1 accessions were collected from geographically (coastal regions of Spain and Japan) and genetically [21] distinct populations. However, both populations contain this new AtHKT1 allele, which makes it tempting to speculate that this allele allows these two populations to thrive in coastal regions where there might be enrichment of NaCl in the soil due to exposure to seawater. However, an extensive haplotype analyses will be required to identify if this new allele of AtHKT1 has been under recent directional selection before we can make any conclusive statements about its adaptive significance.
A. thaliana Col-0, Ts-1, Tsu-1, and other accessions reported in this paper were obtained from the Arabidopsis Stock Center at the Arabidopsis Biological Resource Center. The hkt1–1 mutant [19] was from our own seed stock. For the genetic analysis, F1 and F2 plants from different crosses are as specified throughout the paper.
Plants used for elemental profiling by ICP-MS analysis were grown under unstressed conditions in a controlled environment, 8 h light:16 h dark (90 μmol • m−2 • s−1 light intensity) and 19 to 22 °C [1]. Briefly, seeds were sown onto moist soil (Sunshine Mix LB2; Carl Brehob & Son, Indianapolis, Indiana, United States) with various elements added at subtoxic concentrations (As, Cd, Co, Li, Ni, Pb, and Se [1]) and stratified at 4 °C for 3 d. Plants were bottom-watered twice per week with 0.25× Hoagland solution in which iron was replaced with 10 μM Fe-HBED [N,N′-di(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid monohydrochloride hydrate; Strem Chemicals, Inc., http://www.strem.com). For sodium analysis after 42 d, plants were nondestructively sampled by removing one or two leaves. The plant material was rinsed with 18 MΩ water and placed into Pyrex digestion tubes.
The evaluation of the F2 population from the Tsu-1 × Col-0 cross in response to salt stress was realized in a growth chamber under controlled environmental conditions, 8 h light:16 h dark (150 μmol • m−2 • s−1 light intensity), 21 °C:19 °C (day/night) and 60% relative humidity. After stratification, F2 seeds as well as seeds from each one of the parents were sown onto moist soil (Scotts Potting Medium; Scotts-Sierra Horticultural Products Company, Marysville, Ohio, United States). About 2 wk after germination, seedlings were thinned to leave one plant per individual cell. Plants were bottom-watered twice per week with 0.5× Murashige and Skoog (MS) Macro- and Micronutrients (Caisson Laboratories, Inc., Rexburg, Idaho, United States). One month after the seeds were sewn, NaCl was added to the watering solution increasingly from 50 to 100 mM NaCl and maintained at that level until all the plants were recorded dead.
Plants from Col-0, hkt1–1, Ts-1, and Tsu-1 used to evaluate Na+ accumulation in the shoot in response to NaCl stress were grown under controlled growth conditions (as described above in "General plant growth conditions") and treated with NaCl for 2 wk. NaCl treatment was started 1 mo after the seeds were sewn. Plants were watered twice weekly without or with NaCl. The NaCl treatments included plants watered with 50 mM NaCl and plants watered once with 50 mM, once with 75 mM, and twice with 100 mM NaCl. Two weeks after the beginning of the salt treatments, plants were harvested and analyzed by ICP-MS. Na+ was analyzed in leaves of different age throughout the shoot. The leaves were numbered from the meristem toward the outer leaf ring of the rosette. Four samples (indicated as Nos. 1, 2, 3, and 4 in the graph) were harvested per plant, with No. 1 containing leaves Nos. 4 and 5 and therefore being the youngest leaves, No. 2 containing leaf No. 8, No. 3 containing leaf No. 11, and No. 4 containing leaf No. 14 and being the oldest.
Approximately 3 mg of dry weight of each plant was sampled into Pyrex tubes (16 × 100 mm) after drying at 92 °C for 20 h. After cooling, seven of approximately 100 samples from each sample set were weighed. All samples were digested with 0.7 ml of concentrated nitric acid (OmniTrace; VWR Scientific Products; http://www.vwr.com) and diluted to 6.0 ml with 18 MΩ water. Elemental analysis was performed with an ICP-MS (Elan DRCe; PerkinElmer, http://www.perkinelmer.com) for Li, B, Na, Mg, P, K, Ca, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, and Cd. All samples were normalized to calculated weights, as determined with an iterative algorithm using the best-measured elements, the weights of the seven weighed samples, and the solution concentrations, implemented in Microsoft Excel (http://www.microsoft.com) [1].
DNA microarray-based BSA was realized as previously described [12,13]. Briefly, SFPs were identified between Col-0 and Ts-1 and Tsu-1 by hybridizing labeled DNA from each one of the accessions to Affymetrix ATH1 microarrays and comparing them to Col-0 hybridizations downloaded from http://www.naturalvariation.org/xam. Two genomic DNA pools from an F2 population of a cross between Ts-1 or Tsu-1 and Col-0 were created and hybridized to arrays. Each one of the pools contained plants with either levels of Na+ similar to Col-0 ("control" pool) or high Na+ levels similar to Ts-1 or Tsu-1 ("high Na+" pool). At loci unlinked to the ionomic phenotype of interest, the pools should have equivalent amounts of each genotype, and the hybridization signal at each SFP should be intermediate between the two parent accessions, for an average difference between the two DNA microarrays of zero. At linked loci, the difference between the two DNA microarrays should be approximately two-thirds the difference between the parent accessions. By smoothing the signal across multiple SFPs, the noise is reduced and the peak of the differences in hybridization signal will correspond to the chromosomal region of the loci controlling the mapped trait. Raw hybridization data (.CEL files) for each probe on the ATH1 DNA microarrays used in these experiments have been submitted to both Natrualvariation.org (http://www.naturalvariation.org) and Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo) for public distribution.
To sequence the AtHKT1 gene and corresponding promoter from Ts-1 (CS1552) and Tsu-1 (CS1640), synthetic oligonucleotide primers were designed to produce overlapping amplified PCR products from 5,456 bp upstream of the AtHKT1 start codon to 386 bp downstream of the AtHKT1 stop codon, corresponding to positions 32,115 and 41,615, respectively of the BAC clone F24G24 in Chromosome IV.
Table S1 provides a summary of the pairs of primers used and the size of the PCR products expected for Col-0 (based on published sequence) and obtained for Ts-1 and Tsu-1. The PCR products obtained for each one of the primer pairs were cloned using the pGEM-T Easy Vector System I according to the manufacturer's instructions (Promega, http://www.promega.com). After transformation of electrocompetent Escherichia coli cells strain DH5α and selection of transformed colonies using ampicillin, positive clones were confirmed by PCR. The plasmids containing the inserted PCR product were extracted and used to carry out the sequencing reaction in both directions using Big Dye terminator v 3.0 according to the manufacturer's instructions (Applied Biosystems, http://www.appliedbiosystems.com). The sequencing products were then analyzed at the sequencing center (Purdue University), and sequences were compared to the corresponding Col-0 sequence from the Arabidopsis database.
Plants were first analyzed by ICP-MS and further used to determine the AtHKT1 transcript levels. Shoot and root from plants grown under short-day conditions (as described above in "General plant growth conditions") were separated and rinsed thoroughly with deionized water to remove any soil contamination. The samples were then frozen in liquid nitrogen and stored at −80 °C until extraction. Total RNA was extracted from each one of the samples using the Qiagen RNeasy Plant Mini Kit (http://www.qiagen.com), and DNase digestion was performed during the extraction procedure according to the manufacturer's instructions. Two micrograms of total RNA was used as a template to synthesize first-strand cDNA with random hexamers using SuperScript II Reverse Transcriptase (Invitrogen Life Technologies, http://www.invitrogen.com). Quantitative real-time PCR was performed with the first strand cDNA as a template on a sequence detector system (ABI Prism 7000, Applied Biosystems). For normalization across samples, the expression of the Actin 1 gene (At2g37620) was used with the following primers: CPRD66, 5′-TGG AAC TGG AAT GGT TAA GGC TG-3′ and CPRD67, TCT CCA GAG TCG AGC ACA ATA C-3′. For AtHKT1, the following primers were used: HKT-RTF, 5′-TGG GAT CTT ATA ATT CGG ACA GTT C-3′ and HKT-RTR, 5′-GAT AAG ACC CTC GCG ATA ATC AGT-3′. For each sample, the average value from triplicate real-time PCRs was used to evaluate the transcript abundance, and the relative fold induction relative to AtHKT1 expression in Col-0 shoots was calculated based on the ΔΔCt method.
Seedlings to be grafted were germinated on 100 × 15-mm plates containing 0.5× MS Macro- and Micronutrients, 0.5× MS Vitamins (Caisson Laboratories, Inc.), 3 mg/L Benomyl [methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate; Sigma, http://www.signaaldrich.com), 0.04 mg/L BA (6-benzylaminopurine; Sigma), 0.02 mg/L IAA (indole acetic acid; Sigma), and 12 g/L agar. Hormone treatment was found to greatly improve grafting efficiency due to at least three factors: enhanced callusing at the graft union, retardation of shoot growth which maintained contact at the graft union, and an approximately 90% reduction in adventitious root formation from the graft union. Benomyl virtually eliminated fungal contamination [26]. Plates containing the stratified seeds were placed vertically under controlled environmental conditions (16 h light:8 h dark and 25 °C). Five-day-old seedlings were grafted on the plate by the 90-degree blunt end technique with a 15-degree Stab Knife (Fine Scientific Tools, North Vancouver, British Columbia, Canada) without collars [27]. The grafted seedlings remained on the plate for an addition 5 d to allow the formation of the graft union. Successfully unified seedlings were transplanted directly to soil (as described above in "General plant growth conditions"). Plants were harvested for Na+ analysis 4 wk after transfer to soil. Postharvest analysis of graft unions was performed under the stereoscope to identify any adventitious root formation from grafted individuals. Individuals with adventitious roots emerging at or above the graft union were eliminated from subsequent analyses.
Na+ contents were analyzed in leaves at four different stages of development (as described in Materials and Methods) across the shoot of 44-d-old Col-0 (circles), Ts-1 (triangles), Tsu-1 (squares), and hkt1–1 (diamonds) plants grown in soil under short-day conditions and treated for 17 d with (A) no NaCl addition, (B) 50 mM, or (C) 100 mM NaCl. Presented data are the mean ± SE.
The Arabidopsis Biological Resource Center (ABRC) (http://www.biosci.ohio-state.edu/pcmb/Facilities/abrc/abrchome.htm12) accessions used in this paper were Col-0 (CS6000), Cvi-0 (CS1096), Est-1 (CS1150), Kas-2 (CS1264), Mrk-0 (CS1374), Mt-0 (CS1380), Se-0 (CS1502), Ts-1 (CS1552), Van-0 (CS1584), Nd-1 (CS1636), Tsu-1 (CS1640), and Ler-2 (CS8581).
We would like to thank the Arabidopsis Biological Resource Center for providing the seeds of the Arabidopsis accessions used in this paper.
Competing interests. The authors have declared that no competing interests exist.
A previous version of this article appeared as an Early Online Release on October 26, 2006 (doi:10.1371/journal.pgen.0020210.eor).
Author contributions. AR, IB, and DES conceived and designed the experiments. AR, IB, BM, JG, BL, and EY performed the experiments. AR, IB, BM, JG, BL, and DES analyzed the data. AR and DES wrote the paper.
Funding. This research was supported by the National Science Foundation Plant Genome Research (DBI 0077378) and Arabidopsis 2010 (IOB 0419695) programs and the Indiana 21st Century Research and Technology Fund (912010479).
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


text:
Contact Us
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For hours of business, our telephone and fax numbers, mailing addresses, ATM information and location maps:
Report a lost or stolen Credit Union Mastercard or YNCU Debit Card
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We are dedicated to the highest levels of member service and satisfaction. Like all credit unions, we are subject to regulation by the Credit Union and Caisses Populaires Act and must adhere to accepted practice. If you have a concern about a matter regarding your dealings with Your Neighbourhood Credit Union, and it cannot be settled with your branch, you can contact the Complaint Resolution Officer using the Online Contact Form. If the Complaint Resolution Officer is unable to provide a response that is satisfactory to you, the Officer will provide direction on the next steps available to you.
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Please allow 1-2 business days for an email response. Should your matter be more urgent, or if you need to provide personal information for us to assist you, please contact us by telephone.

subset: commoncrawl


text:
Contact Info
People Helping People Agency - Polet Bet-David
Polet Bet-David
28422 Constellation #118
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Phone: (661) 622-3455
Contact via Email
Visit our Website
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Patrick Bet-David is a man who inspires. He is a successful entrepreneur, emerging author and popular blogger committed to helping people get more out of life. Patrick takes complex ideas and converts them into simple life lessons.
His intriguing perspective on opportunity is colored from his experience as a young immigrant from war-torn Iran in the late '80s. He has a true rags-to-riches story that saw him first serving in the U.S. military before breaking into the financial services industry. Before turning 30, Patrick started a PHP Agency, one of the fastest growing companies in the U.S.
As entrepreneur and author, Patrick is shaping the next generation of leaders with a voice of honesty and sometimes controversial insights. In his first book, "Doing the Impossible: The 25 Laws of Doing the Impossible", Patrick gives practical advice on how anyone can take ownership of his or her own life and rediscover dreams and aspirations. In "The Next Perfect Storm", he shares a look at the current market conditions creating the next financial boom.
A regular blogger for over three years, Patrick recently turned a weekly radio show in Los Angeles into a video blog series called "Valuetainment Weekly" where he shares practical advice on all facets of life. He is also the voice behind "Newton's Bark", a weekly finance lesson from GoAskNewton.com, a website designed to help educate people on all things related to personal finance.
Find out more about Patrick and his other accomplishments at www.patrickbetdavid.com


text:
Radikal Therapeutics (RTX) is developing a novel pharmaceutical therapy for prematurity that targets a developmental deficiency in thioredoxin (Trx), a protein that serves as a central regulator of cellular reductant status. Trx is expressed t levels in the fetal lung too low to adequately counter the redox stress of extrauterine life, and i thus relatively deficient in extreme prematurity. The low level of Trx in alveoli is thought to pla a major role in the susceptibility of very low birthweight (VLBW) premature infants to neonatal respiratory distress syndrome (RDS). Our current understanding of RDS and its evolution into bronchopulmonary dysplasia (BPD) is consistent with a mechanism of injury produced by an excess of superoxide within the lung parenchyma. Compounding this increase in superoxide production is the developmental deficiency of anti-oxidant proteins, such as superoxide dismutase and Trx, that are present in insufficient quantity at an early gestational age. Our innovation is based upon the administration of a novel agent, R-908, a prodrug of a Trx mimetic (R-901) that substitutes for the deficient Trx protein and restores intracellular redox status. R-901 is a thiol-rich tripeptide closely analogous to the native conserved Trx motif and exhibits extraordinary potency: R- 901 is 450-fold more potent than N-acetyl cysteine in the protection of cultured cells from oxidant stress. In a murine ovalbumin model of pulmonary inflammation model, R-901 reduced histologic injury, diminished neutrophil infiltration, attenuated tissue oxidation, blocked pro-inflammatory cytokine expression and nuclear translocation of NF-?B, diminished the degradation of the anti-inflammatory cytoplasmic protein I?B?, and restored the balance of reduced and oxidized forms of glutathione. In a murine LD60 model of redox stress, induced by acute Cl2 inhalation, post-insult administration of R-901 eliminated all mortality. To overcome shelf instability of the free thiol groups of R-901, we have invented a stable dithioester prodrug (R-908) that releases R-901 in vivo. R-908 will now undergo evaluation in a clinically-relevant rat pup model of BPD.
Aim #1 : Define the pharmacokinetics (PK) of R-908 in newborn rats. We will define the plasma and tissue PK profile of R-908, and its metabolite R-901, in 2-day old rat pups.
Aim #2 : Establish that R-908 attenuates changes of pulmonary vascular and alveolar structure in a hyperoxic model of BPD in neonatal rats;2-day old rat pups will be subjected to hyperoxia for 10 days. R-908 will be administered over a broad dose range for 3 weeks, a period characterized in this model system by progressive lung fibrosis, pulmonary arterial hypertension (PAH), and hypoalveolarization. Lung tissue taken at necropsy will be analyzed for pulmonary vascular structure and growth, alveolarization, lipid peroxidation (malondialdehyde), reduced and oxidized glutathione (GSH, GSSG), peroxynitrite formation (3-nitrotyrosine), poly(ADP-ribose) formation, fibrosis (Mason Trichrome staining for collagen), R-908 and R-901 levels, and pro-inflammatory gene expression. The heart will undergo morphometric analysis for evidence of PAH (as shown by right ventricular (RV) hypertrophy).
Premature birth is frequently associated with an acute respiratory disease that may convert to a long-term crippling lung impairment, known as bronchopulmonary dysplasia (BPD). There are no specific existing therapies that can reliably block the development of BPD. We are developing a novel drug that targets the basic mechanisms of this condition and will test this agent in a clinically-relevant animal model of premature lung disease.


text:
Hello there BertyG I noticed your post as you were wanting to buy a gaming PC. I will be buying one too in a few months time and it's also been a few years since I bothered with gaming - we will be spoilt for choice that's for sure!
My last PC was bought from Scan, the company you mention in your post. It sounds like you want to build your machine yourself (something I could never attempt). I unfortunately am no pro when it comes to PC parts and like you, also don't know much about certain bits inside my machine. I do recommend you perhaps base your build on one of the cheaper gaming PCs which they have pre built though, for example, the £630 3XS Custom Gamer 5i & then maybe customise any bits if you need too. Then try to buy the parts as oppossed to the pre built version.
GXF card. £220 for a 660ti seems excessive. Amazon £170, 50 quit to spend on something else. Shop around if you're truely budgeting :)
SSD. I have an 80gb, containing windows 7, photoshop and a few essential programs. MP3s, photography go on either my standard 500gb hard disk or my external 2tb passport drive. I'm not sure I would put games on an external drive...I just don't know about this? I could if I wanted, install a few games on my SSD and then uninstall them once finished. Maybe you could too?
Anyhow, hopefully someone else will pop by and give a bit more help, all the best with your build.


text:
The Baseline Diet 2009: Part 1
Next is a series of questions: How many meals are you eating per day? How many calories? How many grams of protein? Carbs? Fat? When's the last time you ate fruit or vegetables? How much water are you consuming on a daily basis. If you're an average lifter (and want to stay such), your answer is probably 'Umm, I don't know.'
General Philosophies of Muscle Mass Gain
In this article (which will actually form an introduction to a series of articles I'll be doing over the next several weeks and months), I want to talk about some basic concepts related to mass gaining nutrition, primarily looking at some of the different philosophies of mass-gaining that are out there. As usually, I'll look at each in my normal way, looking at the various pros and cons of each approach.
Muscle Gain Mistakes
Although it may seem strange to talk about how to gain weight as we approach the holidays (where people typically gain weight without trying very hard), the simple fact is that, for athletes and bodybuilders, the winter (when it's cold outside and you're covered up) has always been one of the primary times that trainees focus on muscle gain.
Contest Dieting Part 1
Of all athletes in the world, bodybuilders (and other physique oriented folks such as fitness and figure girls) tend to be the most anal compulsive and neurotic about their food intake. Nowhere is this seen more than during contest dieting where folks that are already on the far edge of what most would consider sane turn batshit crazy about their food intake.
Initial Body Fat and Body Composition Changes
In that current data indicates that each pound of muscle might burn an additional 6 calories (as opposed to older values of 25-40 cal/lb or even higher) (1), this argument is no longer tenable; to significantly affect metabolic rate would require a monstrous gain of muscle mass, far more than you could gain in 3-4 weeks.
Periodization for Bodybuilders: Part 3
I finished the second part by giving some volume recommendations for both training and maintaining loads for the different components (4 of them) of training: pure strength, intensive bodybuilding, extensive bodybuilding and really extensive bodybuilding. Without recapping that entire article, I'll simply summarize the loading parameters for each below.
Periodization for Bodybuilders: Part 2
In his original holistic training schema, Dr. Hatfield proposed using three different intensity/rep ranges to optimally stimulate a muscle. This included sets of 4-6 done explosively, sets of 12-15 done rhythmically and sets of 40 done fairly slowly. Different types of workouts were done in a fairly complicated cycling pattern (Hatfield called this ABC training) and, frankly, keeping everything straight was a huge pain in the ass.
Periodization for Bodybuilders: Part 1
Periodization is one of those terms that gets thrown around a lot and can have many many many meanings, depending on who you're talking to. From fairly generic approaches to cycling training to meticulously planned out programs where ever set and rep is set ahead of time, you can find many different intrepretations of periodization and what it means. In this article series, I want to discuss periodization as it applies to bodybuilding specifically.


text: General Dynamics NASSCO, a subsidiary of General Dynamics announced the start of construction on the fifth dry cargo-ammunition ship in the U.S. Navy's T-AKE program. The ship will be named later and is scheduled to be delivered to the Navy in the first quarter of 2008. NASSCO has been awarded contracts to build nine T-AKE ships. Two additional ships are expected to be ordered by the Navy over the next two years for a total class of 11 ships. The first ship of the class, USNS Lewis and Clark , was delivered to the Navy on June 20. The second ship, USNS Sacagawea , was launched on June 24 and is scheduled to be delivered in the first quarter of 2007. The third ship, USNS Alan Shepard , is scheduled to be launched on December 5, 2006, and delivered in the summer of 2007. The keel laying of the yet-to-be-named fourth ship will take place on July 28, 2006. The T-AKE ships are 689 ft. in length and 105.6 ft. in beam, with a design draft of 29.9 ft. and a displacement of 41,000 metric tons. NASSCO has incorporated international marine technologies and commercial ship-design features into the ships, including an integrated electric-drive propulsion system, to minimize operating costs over their projected 40-year service life.


text:
FIFA bans Chuck Blazer from football for life
Johannesburg: Former US football official Chuck Blazer has been banned from all-football related activities for life on Thursday by FIFA over his involvement in corruption and bribery scandals.
FIFA said in a statement that Blazer committed many and various acts of misconduct continuously and repeatedly during his time as an official in different high-ranking and influential positions at FIFA and CONCACAF, Sport24 reported.
The 70-year-old worked undercover with US prosecutors in order to expose illegal practices at FIFA, after pleading guilty to charges of bribery, money laundering and tax evasion.


text:
- Description
- Data Sheet
- Suitable for (14)
- Reviews
DetailsLarger carbs supply larger quantities of fuel-air mixture. As the incoming mixture is considerably cooler than the combustion chamber this means better internal cooling of the entire system. And it also means more power. With a larger carb you increase both, power and reliability. Our carbs are supplied with the correct jetting, though sometimes playing around with the mainjet may be necessary. In most cases the original airfilter may still be used.
Data Sheet
Name Carburettor 17.5 mm Item No. L-DEL1438 Brand Dell'Orto Engine Type 2-stroke Carburator Type Dell'Orto Carburator Diameter 17,5mm Slide type Round
Your product goes with the following manufacturers and their models:
Bali 50cc AC - Honda
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X8R-S 50cc AC - Honda
Bali 50cc AC
SFX 50cc AC
SGX SKY 50cc AC
SH 50cc AC (after 1996)
SXR 50cc AC
X8R-S 50cc AC
X8R-X 50cc AC


text:
37320RE: Lindvay update - more confused
- Nov 15 8:29 PM
In reply to your extended query regarding Lindvay.
What you are finding is somewhat more complicated than many researchers, but not at all unusual. Remember that death certificates, tombstones and census records in the U.S. are often in serious error since an inquiring officer just writes down what someone under stress reports to them. Oftentimes, the person reports from bad memory of just doesn't know and guesses because they feel obligated to answer a question. And on occasion they purposely decieve for reasons we can only guess. Among immigrants there were oftentimes problems of language and understanding as well.
The records that are almost always exacting and correct are the records of birth and baptism with place and the reporting of parentage. You have almost certainly found the correct birth records.
First name changes are frequent upon immigration to the U.S. Norbertus to Albert, and Franciscus, or more likely Frantisek to Frank are common.
The reporting of Frank as brother to Albert in the census is probably just an error. Perhaps the census taker and reporter didn't even speak the same language. Frank and Albert are much more likely both children of fathers who are cousins, but resided in the same family home in Rudnik. Their more distant relationship would not be an impediment to marriage with Albert's daughter Maria.
Exactly, what is the town near Kosice you can't find where you mention the marriage took place?? Earlier you said they were married in Kosice.
Curt B.
---In email@example.com, <nina.smith.m@...> wrote:I posted a few weeks ago about my problems with Lindvay research. I've just
found new info which has actually made me even MORE confused.
My ggg grandparent's came from around Kosice, Hungary (now Slovakia) to Van
Meter, PA in the late 1880s. Their names were Albert Lindway and Maria
(Bernat)... or WERE they?
Albert died in 1907 in the Darr Mine diaster which killed over 200 men. His
headstone and death certificate state that he was 47 years old when he
died. No birth date or year was written anywhere. The census data has
various birth years written for him, from 1861 to 1864. His headstone is
written in Slovak.
His wife Maria supplied the info for Albert's death certificate. Maria came
to America after Albert, and had their baby, also named Maria, about 1 yr
old, with her. I have yet to find any immigration records for either of
The parents listed on Albert's death certificate are "Andy Lindway" and
something that looks like "Kovck Krencrika."
In the 1900 census, there is another person living with Albert and Mary and
their children, Frank Lindway, listed as Albert's brother, but 20 years
younger. This Frank went on to marry Albert and Mary's daughter, Mary, in
1904. If we are to believe the census, this would make Frank young Mary's
own uncle. I am waiting on Frank's death certificate in hope's that
parent's names are listed. Frank died in 1913.
I recently discovered a baptism record in Rudnik, Slovakia (close to
Kosice) for a NORBERTUS Lindvay, born Aug 1860. Parents are GEORGIUS
Lindvay and Franciska Kolvek.
I also found that this Norbert went on to marry a Maria Bernath in Kosice
in 1886 which is when Albert and Mary state they were married in the US
I also found that this Norbert and Maria had a child, Maria, in Kosice in
Feb 1887 which is when I know Mary Lindway was born, based on US records.
So I had strong evidence to believe that Norbert was actually my Albert.
Why would he change his name from Norbert to Albert?? And why is his
father's name written as "Andy" when if it was actually Georgius? It would
also mean that he was 48 years old when he died, and not 47 as it says on
his headstone and death certificate. So I still wasn't sure...
Today I got an email from a priest in PA who sent me a picture of the death
register of the church that Albert and Mary attended in PA. It showed
Albert's death and listed his place of birth as Rudnok (now "Rudnik"). I
finally had a city for him, which matched up again to that Norbert baptism
record. But this record again listed Andreas as Albert's father, not
Georgius. This info was probably provided again by Mary. How could she not
know the father of her husband? But this record listed his mother's name as
Francisca Kolvek, which again matches the Norbert record.
I have looked through the Rudnik baptisms from 1859--1865 and there are no
Albertus Lindvays and it is a very small town.
I have looked at all the children's names that Georgius and Franciska had
(13) and nothing close to "Albert."
Now it gets even more confusing. Frank Lindway, the 20-year-younger brother
of Albert... Was he really a brother? 20 years is a big difference.
Especially since Albert was one of the last of 13 children. I thought maybe
the mother died and the father remarried and Frank is really a
half-brother. I am still waiting on Frank's death record and have also
asked the same priest if he can find anything in that registry.
But I just found a baptism record in Rudnik, IN THE SAME HOUSE that Norbert
was born, for a Franciscus Lindvay in May 1881, which is when I know Frank
Lindway was born, based on US records. His parents however, are NOT
Georgius and Franciska. They are listed as Andreas Lindvay and Maria Kozel.
But they are living in the same house that Norbert was born. Who are these
people? Why does Albert claim this Frank to be his brother if neither of
Frank's parents are Albert's parents? I thought maybe, somehow, Georgius
was Andreas (which would explain why Andreas is written on the death record
for Albert) but I looked up the marriage of Andreas and Mary and they were
married in 1869 with him only being 22 years old. So it is not possible. I
thought maybe Andreas was a sibling of Albert (making Frank a nephew),
which would explain why he is living in the same house, but they did not
have any children names Andreas or anything remotely similar.
Can anyone make sense of this?
Also, does anyone know the city listed in this marriage record for Maria
Bernath? It should be located around Kosice but I am not finding it on a
Thanks for any help!! It is much appreciated.
[Non-text portions of this message have been removed]
- << Previous post in topic Next post in topic >>


text:
So its Saturday early evening and I am listening to music.
Gives me hope for monotone!
COPS is about to come on soon. I love COPS, I watch because I look forward to the possibility of seeing people I know running from the PO-PO. Living in North Carolina, New Mexico and Kentucky, you sure do meet a lot of interesting folks who will do anything to become famous.
My hats off to Sucka for pulling off a brilliant first of April prank. You are one of the only in DC to have an actual sense of humor, The Saviours be damned.
My hats off to Junebug McQueen for her most excellant first of April prank. All I can say is we're expecting around October 31. Happy Halloween!!
Never got into Helium, but Lucy is amazing!
Saturday, April 02, 2005
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text:
The District of West Kelowna is reminding business owners that business licence renewals are now due. Business licences for 2013 expired on December 31 and business owners have until Friday, January 31, 2014 to secure their 2014 licence before late fees are applied.
Any business operating in West Kelowna requires a business licence. The District mailed renewal notices to its 1,621 existing business licence holders in November 2013.
Businesses that did not receive their renewal notices or new businesses requiring a licence can contact the District's Business Licensing Division at 778-797-8820.
More information on the District's business licensing services can be found on the District's website www.districtofwestkelowna.ca under Business/Business Licences.


text:
Written on: 20/08/2012
cat treated with advocate administered by vet Beechwwod vet Woodley 6.8.12. came to me 4 hols 16.8.12. found fleas in house next day. checked cat - ridden with fleas. beechwood vet said do not give another treatment, spray house. what is the point of spraying house if cat still has live fleas? have never had probs with other product such as frontline. will see local vet asap
As rated by our community of reviewers
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text:
Roses Coloring Pages
Roses Coloring Pages
On this website we recommend many images about Roses Coloring Pages that we have collected from various sites of coloring page, and of course what we recommend is the most excellent of design for Roses Coloring Pages. If you like the design on our website, please do not hesitate to visit again and get inspiration from all the Free Printable Coloring Pages Images For Kids of our web design.
And if you want to see more images more we recommend the gallery below, you can see the picture as a reference design from your Roses Coloring Pages.
Thank you for seeing gallery of Roses Coloring Pages, we would be very happy if you come back.
Roses Coloring Pages #1
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text:
Nokia Lumia 638 for Rs 5999 from Amazon
Amazon offering you a deal of nokia lumia 638 smartphone at just Rs 5999 online in India.
- 5MP primary camera, auto focus, 4X zoom, video frame rate 30 fps, video resolution 720p (1280 x 720), video recording format MP4/H.264. Camera Apps: Bing vision, Lumia Camera, Lumia Cinemagraph, Lumia Panorama, Lumia Refocus, Lumia Selfie
- 4.5-inch FWVGA IPS ClearBlack LCD capacitive touchscreen with 480×854 pixels resolution, TrueColor(24-bit/16M)
- Windows 8.1 operating system with Lumia Cyan, 1.2GHz Snapdragon 400 quad core processor, 1GB RAM, 8GB internal memory expandable up to 128GB, Single Micro SIM
- Connectivity: Cat 4 LTE/4G support for bands 3, 7, 38, 39, 40, and 41. Battery: 1830mAH, Talk time GSM: Up to 19.5 hours, 3G: Up to 20.1 hours
Nokia Lumia 638 Specifications:
4.5″ Touchscreen Display, 1.2GHz Quad Core Processor, 1GB RAM, 8GB Internal Memory Expandable upto 128GB, 5MP Camera, 1830Mah Battery & Windows 8.1.
Click here to Buy Nokia Lumia 638: Black | White | More Mobiles


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Friio hangs up boots
By Simon Fudge
Last Updated: 01/01/70 1:00am
Nottingham Forest midfielder David Friio has retired from professional football.
Nottingham Forest midfielder David Friio has decided to hang up his boots as a professional footballer.
The Frenchman, who joined Forest in a £100,000 deal from Plymouth Argyle in February 2005, mutually terminated his contract at The City Ground this week.
It has allowed the 33-year-old to pursue a move home to France in order to set up a football-related business.
Friio ends his playing days after spending less than six years in English football, with five of those years coming at Plymouth between 2000 and 2005.


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Turkish President Recep Tayyip Erdogan says Manbij belongs to Arabs and that the PYD and YPG should leave the area. Separately, Syrian regime forces are closing in on Manbij.
Turkey's next target in its ongoing military operations in Syria will be SDF-held Manbij, President Recep Tayyip Erdogan said on Tuesday.
"Manbij belongs to Arabs, not to the PYD or YPG. This is not their place to stay," Erdogan said.
The YPG is the armed wing of the Kurdish PYD — the Syrian affiliate of the PKK — which is recognised as a terrorist organisation by Turkey, the US, and the EU. The SDF (Syrian Democratic Forces) is dominated by the YPG, which captured Manbij from Daesh last year with US backing.
TRT World's Abubakr al Shamahi has more on the president's statement and what it means.
Erdogan said that Turkey does not plan or aim to stay in Syria.
Regime on Manbij perimeter
Erdogan's remarks came as Syrian regime forces closed in on Manbij after taking several villages from retreating Daesh, reports and a war monitor said on Tuesday.
Syrian regime forces opened a new corridor between two territories held by the YPG, redrawing the map of the conflict near the Turkish border, said the Britain-based Syrian Observatory for Human Rights (SOHR).
The advance brings the regime forces closer to areas held by Turkish-backed opposition rebels.
Turkey and its opposition allies are waging a campaign against Daesh and PKK-affiliated groups in Syria near the Turkish border.
The SDF said the Syrian regime's advance would bring benefits to civilians in the area.
TRT World's Abubakr al Shamahi explains how the regime and SDF forces cooperate in the region.
Elsewhere in Syria
SOHR said six civilians were killed and 21 others injured on Monday in regime air strikes on the rebel-held town of Marat Masrin in Idlib province.
The monitor said that fierce fighting is underway between several rebel factions and Daesh in southern Daraa province for a third straight day.
Last Sunday, Jayish Khalid Bin Walid, a Daesh-linked group in Daraa, launched attacks on opposition-held areas and managed to capture several towns and hills from the rebels who were busy fighting the regime forces in Daraa city.
They said that rebel groups were forced to suspend their offensive against regime forces and redeploy their forces to repel Daesh attacks.
SOHR said more than 65 fighters from both sides have been killed so far.
UNSC vote on draft resolution
The UN Security Council is scheduled to vote on Tuesday on a draft resolution that would impose sanctions on Syria for the use of chemical weapons.
Russia says it will use its veto to block the measure, making it the seventh time that Moscow has resorted to its veto power to shield Syrian regime leader Bashar al Assad.
The proposed resolution drafted by US, Britain and France would slap sanctions on 11 Syrian nationals and 10 entities linked to chemical attacks in the nearly six-year war.
It would also ban the sale, supply or transfer of helicopters and related materiel, including spare parts, to the Syrian regime.
The proposals follow a UN-led investigation which concluded in October that the regime air force had dropped chlorine barrel-bombs from helicopters on three opposition-held villages in 2014 and 2015.
Russian Deputy Ambassador Vladimir Safronkov said on Friday that Moscow would veto the measure because it was "one-sided" and based on "insufficient proof."


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Over 1.8 million nonprofits and charities for donors, volunteers and funders
Causes: Art Museums, Arts & Culture
Mission: Art preservation, presentation & education
Programs: Presentation: during 2012, joslyn art museum had 114,254 people visit the museum to view various exhibitions. The following exhibitions were on display in addition to the permanent collection for the general public during 2012: 25 works of art were acquired by joslyn art museum through gift or purchase. 6 works of art were lent by other institutions to joslyn's permanent collection. 1 special exhibitions were on view at the museum.
education: the mission of joslyn's criss center for education is to equip the general public with skills to enjoy, understand, appreciate, and engage with art of various time periods and cultures. To this end, the center provides interpretive programming in a variety of formats tailored to different learning styles. Specifically, joslyn addresses the needs of a general adult audience university faculty and students, k-12 teachers and students, families, and youth-at-risk. Through guided tours, lectures, gallery talk, outreach programs, classes, camps, programs for teachers, and special events, the center for education strives to provide quality arts education experiences for its diverse constituencies. During 2012, 169,022 people attended tours, lectures, galler talks, special programs for adults, art classes, camps and community events at joslyn.
preservation: in formation since 1931, joslyn art museum's collection now contains more than 11,000 works of art from all over the world antiquity to the present, with a concentration on the 19th and 20th century european and american art. Highlights of the permanent collection include works by lorenzo di credi, titian, el greco, veronese, claude de lorrain, edgar degas, claude monet, pierre august renoir, and camille pissarro. American masters such as albert bierstadt, thomas hart benton, mary cassatt, thomas eakins, winslow homer, john steuart curry, grant wood, jackson pollock, dale chihuly, and george segal are also represented. Joslyn is renowned for its collection of art of the american west and known worldwide for its collection of swiss artist karl bodmer's watercolors and prints documenting his 1832-34 journey to the missouri river frontier with german prince maximilian of wied.


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Thursday, August 29, 2013
Corazón Salvaje: Telenovela Rosa with a Twist
1 comment:
My telenovela also has reversed the gender roles with regard to the "Cinderella" story within the show. In my telenovela, La Tempestad, the female protagonist is actually a high-power executive and the boss of the male protagonist. It is also obvious from the entrada and small intros throughout the show that the two will end up together.ReplyDelete
It seems this might be a trend in recent telenovelas. With the female in a position of higher class, income-level or career position, she might also end up having more power in the relationship. This could be a movement to portray equality among women and men in multiple aspects of culture. This is just speculation, as I have not gotten that far in the telenovela I am watching. We will have to wait and find out what happens in order to analyze this trend more!


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Although traditional SMS still has an edge over new messaging applications, it's undeniable that this almost obsolete way of communicating is losing its following at a steady rate. Its most notable advantage is the fact that it is able to send messages across all networks, regardless of the mobile application used. Just like e-mails, SMS can cross platforms and service providers.
But why then, are customers preferring Internet-based messaging? Why are people these days asking for stuff like kik usernames instead of actual phone numbers?
There are several explanations for this. One is that people don't really mind installing numerous applications on their mobile phones. With ample storage capacity and the ability to switch messaging apps through simple swipes, it's really not a big issue for most people. Though it might cause some sort of unease for people who want their phones to look neat and organized. A flood of notifications coming from different applications is never something appealing to look at.
Another quite obvious advantage of Internet-based messaging apps is the capability to use advanced emoticons. SMS is capable of the most basic (read: boring) smiley faces but can never really provide the versatility of the new generation of emoticons. Case in point, how could one send a crying emoticon using base alphanumeric characters? Sometimes a sad face just won't cut it. To prove this point further: you can't do a roll-eye using SMS.
It's also undeniable that messaging apps have the upper hand when it comes to integration with other mobile applications outside the context of messaging. SMS doesn't have the advanced capabilities of dedicated messaging apps like spontaneously sending pictures by tapping a camera button within the application itself.
However, it goes without saying that SMS is still here to stay. It has its use, especially when it comes to communicating with other people who refuse to download certain messaging apps.…


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All quotes tagged Legendary
(25 quotes found)
"J.C. Penney is one of the legendary names in American retailing, and I am honored to be part of this great company. I have met with members of the board and top management, and we are all enthusiastic about what we can achieve together. The desire to succeed is extremely strong here, and I am pleased to have been given he opportunity to lead the company."
"I would probably say yes to Real Madrid, ... It's a legendary team that stands above all others and is the target of all players."
"It's an honor to be a part of such a legendary group of stars representing Max Factor, such as Rita Hayworth and Marilyn Monroe, ... I'm so excited to start working on the campaign."
"It helps you confidence-wise any time you beat a legendary coach like Coach Ashley with 900-something wins. You know a team like that will be well-prepared. You just have to match their intensity."
"One of legendary architect Pete Dye's most celebrated courses, Teeth of the Dog is named for a local term describing the island's jagged-edged coral reefs,"
"One of legendary architect Pete Dye's most celebrated courses, Teeth of the Dog is named for a local term describing the island's jagged-edged coral reefs. The Caribbean-bordering par 3s are the Dog's main draw and down on the fifth and the seventh holes, you're so close to the water that you can feel the spray."
"This gives these young coaches a chance to ask questions from the legendary coaches. It's a very useful tool. They will get the opportunity to find out how these programs were built and how to juggle multiple sports or to turnaround a program."


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ASE Ep. 52: Beware of Low-Flying Defense Drones
Durandal so appreciates our killing skills (hey, it's great somebody does!) that he wants us to come back to the Marathon and do some more of it. We'll even have some "help" of a sort, in the form of defense drones now armed with (loaded) grenade launchers. Friendly fire indeed!
This podcast uses:
Aleph One, free and open source versions of Marathon for Windows, Mac and Linux at http://source.bungie.org
Remixes of the Marathon soundtrack by Craig Hardgrove at http://themarathonmusic.com
Subscribe to the YouTube channel for Anger, Sadness and Envy video podcasts:
Subscribe to the iTunes channel or RSS feed for video and audio podcasts:
Music used in this episode:
"What About Bob?" by Alexander Seropian, remixed by Craig Hardgrove (intro music)
"Leela" by Alexander Seropian, remixed by Craig Hardgrove (gameplay background music)
"New Pacific (Reprise)" by Alexander Seropian, remixed by Craig Hardgrove (outro music)
For more information on this level, visit the Marathon Spoiler Guide:
For complete Terminal texts, visit the excellent Marathon Story Page:
There you can read faithful and complete reproductions of all of terminals in Marathon and its sequels, plus Facts and Puzzling Things About all sorts of characters, places, and concepts in the Marathon universe, as well as a forum where stalwart fans continue to argue the finer points of interpreting Marathon's story nearly twenty years after the game's release!


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Resident Evil 6 Cheats for Playstation 3Home / Playstation 3 /
This page contains Cheats for Resident Evil 6 organized by sections for Playstation 3. This game has "Action Adventure" as genre, made by Capcom, released on Oct 2, 2012. If you can't find a hint or secret in our list, then please check this page periodically for the latest updates.
- Developer: Capcom
- Publisher: Capcom
- Genre: Action Adventure
- Release: Oct 2, 2012
- ESRB: Mature
Easter Eggs
Game Walkthroughs
If you know cheat codes, secrets, hints, glitches or other level guides for this game that can help others leveling up, then please Submit your Cheats and share your insights and experience with other gamers.
Please how can I escaped seem to be getting smashed by falling debri can't work out when it says press L+X ?...
How do I beat the gas spewing fat thing that comes out from under the cathedral altar after I unlock the secret passage...
How Do I shoot all the church bells...
How use the skill points in the game ? there's no where to do that...
After I beat Ada's campaign the Lone Wolf title said 19/20, even though I played every single chapter by myself. I was playing online the whole t...
Don't know if anyone else has noticed but two of the Predator achievemnets are the same. Get Last Place In Predator. I also have 2 different scor...
I only have 2 agent hunt medals to get and not sure how to get them. How do you do the dynamite punch and which enemy do you use? Which enemy spr...
Stuck in this game? Set a specific, clearly defined question and let other gamers answer your question or view answers to previously asked questions. If you think you are an expert then please try to help others with their questions.
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text:
Animal Review 2: Question Preview (ID: 15110)
Below is a preview of the questions contained within the game titled ANIMAL REVIEW 2: Animal Review 2 .To play games using this data set, follow the directions below. Good luck and have fun. Enjoy! [print these questions]
Play games to reveal the correct answers. Click here to play a game and get the answers.
Which of the following is in order from the least complex level to the most complex level?
a) cell, system, organ, tissue
b) cell, tissue, organ, system
c) tissue, cell, organ, system
d) tissue, system, organ, cell
Which two body systems are needed for movement
a) muscle and circulatory
b) endocrine and lymphatic
c) muscle and nervous
d) endocrine and muscle
Which of the following is best described as a structure consisting of several tissues?
a) an organ system
b) an organ
c) a cell
d) an organism
Which two body systems remove foodstuff waste from our bodies
a) digestive and urinary
b) endocrine and lymphatic
c) lymphatic and digestive
d) circulatory and digestive
Which of the following is not a characteristic of arthropods?
a) compound eye
b) eight legs
c) exoskeletons
d) segmentation
The word arthropoda means
a) "jointed leg"
b) "most diverse"
c) "fused body"
d) "exoskeleton"
Of all the animals, what percentage of species are invertebrates?
a) 95%
b) 75%
c) 15%
d) 5%
All of the following are arthropods except
a) spiders
b) crabs
c) clam worms
d) centipedes
The most diverse group of animals on Earth are the
a) reptiles
b) insects
c) mammals
d) amphibians
Millipedes feed mainly on
a) decayed plants
b) insects
c) crustaceans
d) soil
Play Games with the Questions above at ReviewGameZone.com
To play games using the questions from the data set above, visit ReviewGameZone.com and enter game ID number: 15110 in the upper right hand corner at ReviewGameZone.com or simply click on the link above this text.
To play games using the questions from the data set above, visit ReviewGameZone.com and enter game ID number: 15110 in the upper right hand corner at ReviewGameZone.com or simply click on the link above this text.
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text:
A4 Folders Collection Cabinet - Chinaprice: contact company for price
Carbinet Size :278W*340D*878H(mm)
Packing Measurement:41W 34D 95H(CM)
I.Office Product Series
Great Variety:File cabinets with lock, file cabinets
without lock, and transparent and milky white file drawers in
various specification, sizes and types are available, featured
by elaborate design and simple line to build a practical,
efficient, and ideal office environment for you.
Convenient Management:Classification by four-color tag
paper makes management colorful; the dustproof tag shields can
make numbers and words distinct, clear at one glance and
convenient for replacement.
Antiskid Device:Special antiskid design can prevent
files from slipping off when files are drawn out.
Company Contact:
- Phone: 18903039938
- Address: Feida RD Zhangyang Jinyang Industrial Zone523737, DONGGUAN, China
- Email:
- Website: http://WWW.YOUTER.COM
Published date: November 26, 2013 -
- Business Description: Dongguan Youter Industrial Equipment Co., Ltd., Zhangmutou Electronics
City, Youter, File Cabinet, Part Cabinet, Universal Combined Tank,
Document Cabinet, Industrial Arrangement
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* FTSEurofirst 300 up 0.5 percent
* Draghi hints at QE, Euro zone banks gain
* Merck gains after Pfizer deal
* H&M boosted by forecast-beating sales
* Weir hit by renewed slump in crude (Recasts, adds quotes, detail)
By Alistair Smout
EDINBURGH, Nov 17 (Reuters) - European shares turned solidly positive on Monday after ECB President Mario Draghi reasserted that he was ready to do more to fight deflation.
Shrugging off worries that Japan's economy has unexpectedly slipped into recession, the market also drew confidence from earnings reports, merger and acquisition news and appetite for more volatile stocks.
The FTSEurofirst 300 index of top European shares had risen 0.5 percent to 1,351.65 points at 1544 GMT, extending gains after the European Central Bank President said unconventional measures could include the purchase of sovereign bonds.
Andrew Milligan, head of Global Strategy at Standard Life Investments, said ECB action could prompt a change in his "neutral" rating on European equities.
"If we feel the ECB is beginning to surprise the markets with more action, then we probably would look at rotating more into European equity," Milligan said.
"If we felt the ECB were falling further behind the curve, we may have to consider an "underweight" rating."
Euro zone banks extended gains after Draghi's comments, up 1.7 percent, with Unicredit and Banca Monte dei Paschi di Siena both up over 3 percent.
While Monte Paschi said at the weekend no bids had come in for the troubled lender, traders said that appetite for more volatile, or "high beta" stocks was building.
"Traders are betting on big swings in the most hammered European stocks such as (Monte Paschi)... The stocks have such big betas that it's an interesting trade in such a range-bound market," Saxo Bank trader Pierre Martin said.
Pharmaceutical firm Merck rose 3.7 percent, the index's top riser, after agreeing a deal with Pfizer for sharing rights to develop its experimental immunotherapy drug with the U.S. drugmaker.
Veolia also rose strongly, up 2.5 percent, with traders citing a discussion on CNBC on Sunday of new evidence that the earth's groundwater is being pumped out much faster than it can be replenished.
Traders said that this could increase the interest in companies with expertise in water management such as Veolia.
The moves helped to counteract sentiment dented by Japan's drop into recession, with the economy shrinking an annualised 1.6 percent in July-September following a 7.3 percent drop in the second quarter.
Analysts polled by Reuters had expected 2.1 percent growth in the third quarter.
Japan's data hit Brent crude, down more than $1 to near $78 a barrel, further pressuring shares in European oil majors and oil services groups.
Top faller was Weir Group, which dropped to a 2-1/2 year low. It was down 3.7 percent after a double-downgrade from Exane, who warn that the current oil price could trigger 25 pct reduction in rig count. (Additional reporting by Blaise Robinson; Editing by Ruth Pitchford)


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Template >> Flyer - Swimming Pool Cleaning service flyer 246678 by Leza @ CreativeMarket
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What is an Auto Bid?
An Auto Bid in the online auction is the equivalent to an Absentee Bid in the live auction. The simple difference is that the online program will bid on your behalf ONLY if someone else bids below your maximum (or Max Auto Bid), whereas in the live auction there is an Absentee Bidder who carries out your Maximum Absentee Bid under the same circumstances.
What is happening when I place a bid online and immediately am notified that I am out bid?
This is simply the Auto Bid application doing it's job. It does not mean that each time you bid you will be out bid. It simply means that someone else has already left an Auto Bid for a higher value. Once your bid is accepted as the current bid, you now know that you are the highest bidder as of current.
How do I place an Auto Bid?
When you view a particular lot, click the Place Bid button to open a new window with all information on that lot. Then click Auto Bid to open another window. You can either choose your maximum bid from the pull down menu, or enter your maximum bid in the box below that. Enter your paddle number and password, then click Send Bid. You can view your bids as well as your auto bids in the My Bids section of the Online Auction.
Bidding Increments
What if I don't live in Calgary, Alberta, or even Canada?
Levis is obliged to provide clients with various options for packing and shipping as well as accommodating special requests, within reason. All packing and shipping is done at the expense of the purchaser, ie. you. If you are the successful bidder on one or more lot(s), you will receive email notification as such. Please contact us regarding your packing and shipping preferences.


text:
Israel News
Middle East
American Politics
Edition française
Green Israel
Deputy Prime Minister Dan Meridor 311 (R).
(photo credit:David W Cerny / Reuters)
Intelligence Agencies Minister Dan Meridor's wife Leora was
lightly injured Saturday evening during a car accident on Hameyasdim
Street in Jerusalem.Meridor, who was in the car with her, was not injured, Army Radio reported. His wife was taken to Hadassah Ein Kerem Hospital in Jerusalem for treatment.
By DANIEL K. EISENBUD
By LAHAV HARKOV
Sponsored Content
By DANIEL K. EISENBUD
By JEREMY SHARON
Please insert a valid email address


text:
Celebrating Purim with Homemade Hamantaschens
Today marks the Jewish holiday of Purim, a celebration of the story of Queen Esther, who thwarted a plot to eradicate Jews from ancient Persia.
To celebrate, Jews today read from the Megillah (book of Esther); participate in charity; exchange gifts of food; and have a feast for family and friends.
That includes Hamantaschens, delicious fruit-filled pastries shaped like a small three corner hats.
Local couple Levi and Mushky Dubov dropped by the Fox 2 News Studios Sunday morning to tell us more about the holiday and its traditions, as well as how they make Purim's signature snack.


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Costa Rica on a Shoestring - Day 3: Cahuita
Cahuita Travel Blog› entry 24 of 31 › view all entries
July 23rd, 2009 – by: daisywho
I headed back to the hotel and told everyone about the cool turtle mating encounter. After breakfast, we headed to the port and loaded into the boat. The boat ride took about 3 hours and we were able to see lots of birds, reptiles, and other animals. The water in this section of the canal was a lot clearer, it was like a mirror. We managed to get stuck in a shallow area of the canal and our captain had to maneuver us out. There was a big earthquake a few years ago that ruined the canal passages. They have a sediment boat that empties section of the canal so that boats can continue to pass through. Our guide pointed out several different birds along the canal: rojo spoonbill bird, king vultur, and many more I can't remember.
We reached the port and transferred into a bus that drove us to Cahuita. It was a beautiful ride along the coast which took about 30-60 minutes. We checked into our hotel and headed to lunch. After lunch, we changed into our bathing suits and headed into the Cahuita National Reserve. We each gave a donation and entered the park. There was a path that goes through the trees and plants growing along the beach. The vegetation create a sort of tunnel/canopy over the path giving a great shade against the sun.
We met up for dinner to celebrate Rich's birthday and watch the soccer game with Costa Rica vs. Mexico. Our guide managed to talk the owner of Pizza n' Love to bring in a TV so that we could watch the game.
Join TravBuddy to leave comments, meet new friends and share travel tips!


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A raw, compelling glimpse into a young man's charmingly ill-conceived scheme to redeem himself in the eyes of his ex-girlfriend. Ron lures Carol back to his apartment under the pretense that he is going to return her favorite mug. When it seems this will be their last moment together, Ron desperately maneuvers to strike a chord with her in his own uniquely heartfelt way. Written by Sabi Pictures
Watch the official I Fucking Hate You 2008 trailer in HD below or find more I Fucking Hate You videos on Vidimovie.


text:
You've heard of Groupon and LivingSocial (maybe you even broke the bank buying those darn coupons). But what about B2bucks.com or BizyDeal.com?
While deal-of-the-day group coupons are a big hit with consumers, it's uncertain whether they'll catch on in B2B. Still, that hasn't stopped several sites from trying.
The concept usually involves a minimum number of purchases needed for a deal to be activated, serving as a sort of insurance for advertisers. This works well for B2C deals—although not without some profitability challenges.
In B2B, you're even less likely to make big money directly from a group coupon offer. And yet, maybe that's not the point.
The value of a social commerce site, after all, is much more social than commercial. Think of it as another marketing channel to reach and engage new prospects.
So, is a deal-of-the-day right for your business? Depends. It's a more logical fit for small- to medium-sized businesses with transactional business models. Group coupons can help fill the pipeline with new customers for equipment parts, design and delivery services, etc.
If you decide to give it a try, do so as a marketing and advertising investment. Check out these sites: GroupPrice.com , OfficeArrow.com and RapidBuyr.com … and let us know how it goes.


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In her free time Keki likes to stay at home. She likes reading novels or watch movies in her free time. Keki isn't that much keen in travelling and doesn't like going out much.
Keki likes Paulo Coelho's writing, and try not to miss any of his books. She had recently finished reading 'Eleven Minutes' a Paulo Coelho book. At the time of the interview she was reading Amar Neupane's Seto Dharti.
Keki also likes hanging out with friends. Keki and her friends mostly end up going to Bouddhanath Stupa. She and her friends find it peaceful and a wonderful place be around.