text:
Entertainment of Thursday, 4 October 2007
Source: GNA
Accra, Oct. 4, GNA - A one-week contemporary painting exhibition to commemorate 30 years of Korea/Ghana Diplomatic Relations opened in Accra on Thursday with a call on both countries to use the occasion to enrich the bilateral relations between them. The relationship should also generate additional momentum for the expansion of exchanges.
Mr. Wi Keyei-Chul, Ambassador of the Republic of Korea in Ghana, said since the establishment of diplomatic relationship in 1977, both countries had continued to expand and consolidate bonds of friendship, cooperation and partnership in all fields.
He said art was a source of much insight and involved a lot of creativity that depicted a variety of ideas about nature and human beings with its own inherent implications. The exhibition jointly organised by the Korean International Art Exchange Association and the Ghana Association of Visual Artists (GAVA) was under the theme: "The New Vision of Modern Korean Paintings". Mr Keyei-Chul said the exhibition would offer the public the opportunity to see and feel the artistic inspiration and interpretation on nature and life in both Korea and Ghana, thereby promoting a better understanding between the two countries and further deepening the friendship between the people.
"I believe this exhibition by the common language of art, will lay a solid basis for enriching our bilateral relations, and thus will generate further momentum for the expansion of exchanges as well," he said.
Professor Cha Young-Kyu, President of Korea International Art Exchange Association said apart from being part of the anniversary, the exhibition would provide a good opportunity to 'taste' each other's culture in spite of the geographical distance between the two countries. He said they knew Ghana for its preservation of beautiful natural environment and rich traditional culture just as Korea. "Preserving nature means the pursuit of beauty. Only when we preserve our nature well and try to live together with it, (that) we can live a happy life.
"It is exactly the same aim as what most artists eventually want to get through the diverse artistic processes," he said. Prof Young-Kyu said he hoped that the anniversary exhibition would maintain the close relationship between the two countries with the sharing of various cultural and artistic exchanges in addition to diplomatic and economic relations.
Mr. Kofi Osei-Ameyaw, Deputy Minister of Tourism and Diasporan Relations said as paintings depicted culture and values of people, he was of the hope that the exhibition would create further awareness of each other's culture.
He was grateful to the Korean government for its continuous support, especially in science and technology. "We are grateful for affording us the insight into your culture through paintings and I hope that at the end of the exhibition the two associations would learn a lot from each other," he said. Mr. Franklin Nii Glover, President of GAVA, said he hoped that the paintings would give pleasure to the public and thanked the Korean Embassy for the opportunity.
Activities marking the anniversary include painting exhibition, cultural performances, a film festival, street parade, a marathon race, a High School Soccer and Taekwondo Competition.


text: A few years ago I was teaching a three-hour night class at one of the Penn State branch campuses. Half of the way through the class, one of my students tells me he has to leave. I asked him why. He said, as if it were the most natural and obvious reason in the world, that he had to get home in time to catch the season premiere of Two and a Half Men.

subset: commoncrawl


text:
Sarah Larson at the Oscars: Hair and makeup
Sarah Larson's Oscars Look: Love It or Hate It?
George Clooney is such a gentleman. While I'm sure he was hoping that I'd be on his arms at the Oscars, I told him, "George, I wish I could join you, but I'm going to hang out at the Sugar Oscars party." Good thing he was able to find a backup date. Wink.
But seriously now: Here's George and his special ladyfriend, Sarah Larson, who has a bronzed-goddess thing going on. I love her highlighted cheeks, wavy hair, and beautiful smile, but I'm not sure about those spiky lashes. What do you think?


text:
Arsenic is a very potent toxicant. One major susceptibility factor for arsenic-related toxicity is the efficiency of arsenic metabolism. The efficiency, in turn, is associated with non-coding single nucleotide polymorphisms (SNPs) in the arsenic methyltransferase AS3MT on chromosome 10q24. However, the mechanism of action for these SNPs is not yet clarified. Here, we assessed the influence of genetic variation in AS3MT on DNA methylation and gene expression within 10q24, in people exposed to arsenic in drinking water. DNA was extracted from peripheral blood from women in the Argentinean Andes (N = 103) and from cord blood from new-borns in Bangladesh (N = 127). AS3MT SNPs were analyzed with Sequenom or Taqman assays. Whole genome epigenetic analysis with Infinium HumanMethylation450 BeadChip was performed on bisulphite-treated DNA. Whole genome gene expression analysis was performed with Illumina DirectHyb HumanHT-12 v4.0 on RNA from peripheral blood. Arsenic exposure was assessed by HPLC-ICPMS. In the Argentinean women, the major AS3MT haplotype, associated with more efficient arsenic metabolism, showed increased methylation of AS3MT (p = 10−6) and also differential methylation of several other genes within about 800 kilobasepairs: CNNM2 (p<10−16), NT5C2 (p<10−16), C10orf26 (p = 10−8), USMG5 (p = 10−5), TRIM8 (p = 10−4), and CALHM2 (p = 0.038) (adjusted for multiple comparisons). Similar, but weaker, associations between AS3MT haplotype and DNA methylation in 10q24 were observed in cord blood (Bangladesh). The haplotype-associated altered CpG methylation was correlated with reduced expression of AS3MT and CNNM2 (rs = −0.22 to −0.54), and with increased expression of NT5C2 and USMG5 (rs = 0.25 to 0.58). Taking other possibly influential variables into account in multivariable linear models did only to a minor extent alter the strength of the associations. In conclusion, the AS3MT haplotype status strongly predicted DNA methylation and gene expression of AS3MT as well as several genes in 10q24. This raises the possibility that several genes in this region are important for arsenic metabolism.
Citation: Engström KS, Hossain MB, Lauss M, Ahmed S, Raqib R, Vahter M, et al. (2013) Efficient Arsenic Metabolism — The AS3MT Haplotype Is Associated with DNA Methylation and Expression of Multiple Genes Around AS3MT. PLoS ONE 8(1): e53732. https://doi.org/10.1371/journal.pone.0053732
Editor: Brock C. Christensen, Geisel School of Medicine at Dartmouth, United States of America
Received: September 4, 2012; Accepted: December 4, 2012; Published: January 14, 2013
Copyright: © 2013 Engström et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This research was supported by the Swedish Council for Working Life and Social Research, Karolinska Institutet, and Kungliga fysiografiska sällskapet, and the EU (FP6; "PHIME" FOOD-CT-2006-016253). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Arsenic is a widespread environmental pollutant with strong adverse effects in humans, including increased incidence of cancer and effects on skin, respiratory tract, liver, and immune function [1], [2], [3], [4], [5]. Inorganic arsenic is efficiently absorbed in the gastrointestinal tract and is metabolized in the body by a series of reduction and methylation reactions, producing methylarsonic acid (MMA) and dimethylarsinic acid (DMA), both of which are excreted in the urine [6]. In humans, efficient methylation from inorganic arsenic to DMA is associated with a high rate of arsenic excretion in the urine [6]. Incomplete arsenic metabolism, with higher urinary inorganic arsenic and MMA, but lower DMA, seems to be a marker of increased susceptibility to arsenic-related diseases, including cancer [7], [8].
The main methyltransferase in arsenic metabolism is arsenic (+3 oxidation state) methyltransferase (AS3MT), which can methylate both inorganic arsenic and MMA [9], although its affinity for these two substrates may differ. The AS3MT gene is located in chromosome band 10q24, contains 11 exons and spans 32 kilobases [10]. Real time-polymerase chain reaction (PCR) analysis of mature rat tissues detected AS3MT expression in the heart, adrenal gland, urinary bladder, brain, kidney, lung, and liver [9].
Different human populations have varying efficiencies of arsenic metabolism and these efficiencies may reflect underlying genetic differences. For example, compared to other populations, indigenous populations in the Andes demonstrate a uniquely proficient arsenic metabolism, with low urinary excretion of MMA and high excretion of DMA [11]. Moreover, this efficient-metabolizing phenotype was recently shown to be determined by genetic factors; eight single nucleotide polymorphisms (SNPs) (seven non-coding) in AS3MT had strong effects on arsenic metabolism in the population living in the Argentinean Andean highlands, and the efficient-metabolizing haplotype was very common (70%) [12]. Six of these SNPs were also shown to influence arsenic metabolism in people in Bangladesh [12], but the haplotype frequency was much lower (17%) than that in the Argentinean population. All but one of the SNPs in the AS3MT efficient-metabolizing haplotype are non-coding; two of the non-coding SNPs were shown to decrease the expression of AS3MT [12]. Still, the mechanisms for alteration of gene expression are unknown; possibly the haplotype is associated with epigenetic changes in DNA methylation that in turn affects gene expression. The aim of the present study was to assess the influence of genetic variation in AS3MT on DNA methylation and gene expression within 10q24. This work was performed in two distinct human populations, in a subgroup from the above-mentioned population from the Argentinean Andean highlands [12] and a mother-child cohort from Bangladesh.
Background Data, AS3MT SNPs in CpG Sites and Influence on AS3MT Methylation
The characteristics of the different study groups in Argentina and Bangladesh are described in Table 1. In Argentina, only women were analyzed (N = 94 for DNA methylation; N = 90 for gene expression) and their median age was 32 years, they had a median BMI of 24 and 42% reported that they chew coca leaves. Their median total urinary arsenic was 188 µg/L (5/95 percentile 16/620). Their characteristics were similar to the full study population (N = 172) (Table 1). In Bangladesh both newborn boys (N = 62) and girls (N = 65) were examined. Their mothers had a median age of 25, BMI of 20 and their median total arsenic in urine was 68 µg/L (5/95 percentile 20/460).
We next examined the frequencies of the AS3MT haplotype in these two populations and found them to be 69% in Argentina (9% with 0 copies, 44% with 1 copy and 47% with 2 copies – estimated from eight SNPs) and 17% in Bangladesh (71% with 0 copies, 25% with 1 copy and 4% with 2 copies – estimated from five SNPs). The AS3MT haplotype appeared to have a strong effect on the arsenic metabolite pattern in urine, an example in the Argentinean women: increasing copies of the AS3MT haplotype were associated with a higher percentage of DMA (medians 73.0% for 0 copies, 76.0% for 1 copy, and 83.4% for 2 copies; p<0.001 in a general linear model adjusted for U-As). There were no statistically significant differences between the haplotypes in terms of total urinary arsenic (marker of arsenic exposure), age, BMI, and chewing of coca leaves (evaluated in the Argentinean population), or in terms of arsenic exposure during different points during gestation, measured in maternal urine around gestational weeks 6–8 or 30, mothers' age, mothers' BMI, asset score (similar to socioeconomic status), sex of the baby and gestational age (evaluated in the Bangladeshi population) (Table 1).
One potential effect of SNPs on DNA methylation may come from sequence changes that introduce or remove methylation sites; therefore we examined the SNPs associated with the AS3MT haplotype to determine whether these SNPs altered methylation sites and whether these sites were in CpG islands or shores. In the Argentinean women with the major AS3MT haplotype (constituted by rs7085104 G, rs3740400 C, rs3740393 C, rs3740390 A, rs11191439 T, rs11191453 C, rs10748835 A, and rs1046778 C), seven out of the eight AS3MT SNPs are non-coding [12]. The non-coding rs7085104 (5′ of AS3MT) and rs3740400 (intron 1) and the non-synonymous rs11191439 (MetThr exchange at exon 6) SNPs result in new CpG sites (Table S1). The rs7085104 G- and rs3740400 C-alleles introduce CpG sites in the 5′region and the rs11191439 T-allele removes a CpG site. Rs7085104 is situated in a CpG shore 181 basepairs (bp) 5′ of a 424 bp long CpG island, in which rs3740400 is situated. Rs11191439 was not situated in a CpG shore or a CpG island. Therefore, we found that the alleles associated with rapid arsenic metabolism result in the gain of two CpG sites in CpG islands or shores and loss of one CpG site.
We next examined the methylation status of the potential CpG site in rs3740400 C carriers from Argentina by pyrosequencing. An allele-dose increase was found in methylation for the C-allele (comparing individuals of CA or CC genotypes for the rs3740400 allelic site there was significant difference between the groups, p = 0.031; Fig. S1). This result was expected and is an artefact, because for subjects with one CpG (CA) the methylation level would be half of that for subjects with two CpGs (CC). However, we showed with the analysis of rs3740400 that this potential CpG site was differently methylated depending on AS3MT genotype, supporting the idea that non-coding SNPs may render DNA methylation targets. Nevertheless, the degree of methylation was very low (average 3%) in the CC carriers, indicating a small effect on AS3MT gene expression. Due to the LD within and around AS3MT [12], [23], [30], we therefore decided to make a chromosome-region approach and analyzed many CpG sites in 10q24 in relation to AS3MT haplotype. We analyzed the association of the whole AS3MT haplotype on DNA methylation of AS3MT by array analysis (Infinium HumanMethylation 450K BeadChip). In both the Argentinean and Bangladeshi study groups, the AS3MT haplotype was strongly associated with the methylation status of AS3MT (Table 2; Fig. S2). The strongest association was found for the CpG site cg18534077 in the Argentinean study group where AS3MT haplotype was associated with more methylation (r = 0.63, p = 10−6, adjusted for multiple testing). An opposite pattern was found for cg15744005 and cg08772003, where the haplotype was associated with less methylation (Table 2). Only the associations for cg15744005 remained significant in both populations after adjustments for multiple comparisons. Cg15744005 was situated at enhancer elements (Table S1). AS3MT cg18534077 contained a SNP (rs7085104) that could potentially influence base extension (Table 2), although this SNP was situated 27 bp away from the CpG site for cg18534077.
Whole-genome Epigenetic Analysis: Correlation of AS3MT Haplotype on the DNA Methylation Pattern of Chromosome 10
To determine whether adjacent genes are also similarly related to the haplotype, we extended the analysis to an 800 kilobp long region of chromosome 10. In the Argentinean study group, the AS3MT haplotype was strongly associated with DNA methylation of genes located 5′ and 3′ of AS3MT (a region totalling 800 kilobp around the AS3MT gene): CNNM2, NT5C2, USMG5, TRIM8, C10orf26, and CALHM2 (in decreasing order of association, Table 2, Fig. 1, and Fig. S2; full gene names are presented in Table 1). Three of these methylation sites were situated at enhancer elements (Table S1). Three of the top twelve methylation sites (NT5C2 cg09803321, CNNM2 cg11667387, and USMG5 cg18367433) contained SNPs that could potentially influence base extension (Table 2). Two of these (CNNM2 cg11667387 and USMG5 cg18367433) were situated less than 10 bp from the CpG site.
Positions of DNA methylation and gene expression assays are also shown. Black circles denote DNA methylation sites statistically significantly associated with AS3MT haplotype in Argentina and the white circle denotes the DNA methylation site statistically significantly associated with AS3MT haplotype in Bangladesh. A grey box around the DNA methylation ID denotes DNA methylation sites statistically significantly associated with AS3MT haplotype in both Argentina and Bangladesh.
The AS3MT haplotype was associated with more methylation of two sites (one of borderline significance) in CNMM2 and one site in NT5C2. The AS3MT haplotype was also associated with less methylation of one site each in CNNM2, NT5C2, C10orf26, USMG5, TRIM8, and CALHM2. There was a similar pattern in the Bangladeshi population, where AS3MT haplotype strongly influenced the methylation status of CNNM2 (Table 2). The directions of associations (i.e., whether the haplotype was associated with more or less methylation of a specific CpG site) were the same in Bangladesh as in Argentina for the other methylation sites, although these associations were not statistically significant in Bangladesh after adjustments for multiple testing.
The DNA methylation of AS3MT correlated with the DNA methylation of other genes (5′–3′ direction: TRIM8, C10ORF26, CNNM2, NT5C2, USMG5, and CALHM2) within the 800 kb segment of chromosome 10q24 both in Argentina and in Bangladesh (Table S2). We evaluated if the correlation in DNA methylation of genes around AS3MT was particularly strong compared to other regions on chromosome 10, by analyzing windows of 20 consecutive genes and the fraction of CpGs that were correlated between the genes with Pearson correlation >±0.5 (excluding the correlations of CpGs within the same gene). When compared with a permuted (randomized) version of chromosome 10 (green line in plot; Fig. S3) there were some regions that were more correlated than expected. AS3MT (dashed red line) seemed to be in a region with average correlation values.
One mechanism of arsenic toxicity may be through alterations in methylation; therefore, we next tested whether arsenic exposure affected DNA methylation in our samples. We found no clear effect of arsenic exposure on DNA methylation in the studied chromosomal region. In the Argentinean study group, arsenic concentrations in the women's urine were positively correlated with DNA methylation levels of CALHM2 (rs = 0.29, p = 0.012), but not with the other methylation sites that were statistically significantly associated with AS3MT haplotype. In the Bangladeshi study group, arsenic concentration in maternal urine was inversely correlated with CNNM2 CpG sites cg11667387 (rs = −0.23, p = 0.012) and cg00894378 (rs = −0.21, p = 0.019) in cord blood.
Influence of AS3MT Haplotype on the DNA Methylation Pattern of Chromosome 10: Multivariate Models
CpG sites that were statistically significantly correlated (after adjustments for multiple comparisons) with AS3MT haplotype in either Argentina or Bangladesh were further analyzed in univariable and multivariable regression models (Table 3). In total, twelve sites were evaluated; here no further adjustments were made for multiple comparisons. Adjustments were made for total urinary arsenic (natural ln transformed), age and use of coca leaves in Argentina; and for total arsenic in maternal urine (gestational weeks 6–8; natural ln transformed), maternal age and sex of the child in Bangladesh. The results were very similar in the univariable and multivariable models; all sites that were statistically significantly associated with AS3MT haplotype in the univariable analyses remained statistically significant in the multivariable analyses, and the β-values remained the same or changed only marginally.
Whole Genome Expression Data: Associations between DNA Methylation Pattern of Chromosome 10 and Gene Expression (Argentina)
To determine the effect of the altered DNA methylation in the AS3MT haplotype, we also measured expression of AS3MT and adjacent genes in our samples from Argentina. The assay of the AS3MT 3′untranslated region (UTR), NT5C2, TRIM8 and USMG5 showed the largest variation in expression between individuals (Table S3). Expression of all genes associated with the AS3MT haplotype was correlated with each other (Table S4), but not as strongly as these genes' DNA methylation was correlated with each other. For the CpG sites positively associated with the AS3MT haplotype, inverse correlations (Table 4) were found with gene expression of AS3MT (3′UTR assay; p-values≤0.002 or lower for all four CpG sites) and CNNM2 (p = 0.012 for one CpG site; Fig. 2), whereas positive correlations (Table 4) were found with expression of NT5C2 (p≤0.014 for three CpG sites) and USMG5 (p≤0.003 for all four CpG sites, Fig S4). AS3MT cg18534077 was associated with both AS3MT expression measured with the 3′UTR assay (p = 0.002) and with the AS3MT transcript on the array (assay 1771732, p = 0.016).
Scatterplot depicting the relationship between DNA methylation (expressed as fraction of methylated CpG) for CNNM2 (X-axis) and gene expression for AS3MT (3′UTR; expression relative to the median for individuals with 2 copies of the AS3MT haplotype) (Y-axis), where individuals are stratified for AS3MT haplotype (total N = 48). Outliers are denoted by *; the outlier to the left has a relative gene expression of 22.8 and the outlier to the right has a relative gene expression of 24.9.
An opposite pattern was observed for the CpG sites inversely associated with the AS3MT haplotype (Table 4): positive correlations were found with expression of AS3MT (p≤0.003 for three CpG sites) and CNNM2 (p≤0.022 for three CpG sites), whereas inverse correlations were found with expression of NT5C2 (p = 0.038 for one CpG site) and USMG5 (p≤0.002 or lower for all four CpG sites) (Table 4). This was also observed for the CpG sites in TRIM8, C10orf26 and CALHM2 that were inversely associated with the AS3MT haplotype; however, the correlations were less strong (Table S5).
Total arsenic concentration in urine was positively correlated with expression of one transcript of CNNM2 (1754752, rs = 0.23), and negatively correlated with the expression of one other transcript of CNNM2 (1663975, rs = −0.20) and one transcript of AS3MT (1771732, rs = −0.20) (both of borderline statistical significance, Table 4). Only CNNM2 1663975 remained statistically significant in the multivariable analyses (p = 0.021).
Haplotype was associated with gene expression of AS3MT 3′UTR (rs = −0.53, p-value<0.001), NT5C2 (rs = 0.22, p-value = 0.048) and USMG5 (rs = 0.60, p-value<0.001).
Our results show that the AS3MT haplotype, previously associated with a more efficient arsenic metabolism [12], is strongly associated with the methylation status of AS3MT and multiple surrounding genes within a large region of 800 kilobp in chromosome band 10q24 in two different populations. In line with the general concept of CpG methylation functioning to reduce gene expression [13], the carriers of the AS3MT efficient metabolizing haplotype seemed to have reduced expression of AS3MT and CNNM2. However, the increased CpG methylation was also associated with higher expression of NT5C2, C10orf26, USMG5, TRIM8, and CALHM2. Overall, these associations remained stable and statistically significant also after adjustments for other potentially influential variables and show that AS3MT haplotype is the major determinant for the methylation and expression pattern in this chromosomal region. Our finding of correlations in DNA methylation between genes around AS3MT is not unusual, when we compared to other regions on chromosome 10. However, the strong association of the AS3MT haplotype with the DNA methylation pattern, and to some extent to the expression of genes in this region is striking and may reflect the possibility that the genes participate together in arsenic metabolism, or in a not yet characterized pathway.
The strongest association between the AS3MT haplotype and DNA methylation of AS3MT was found for the CpG site cg18534077, which is located in a CpG shore in the 5′UTR of the gene. We evaluated if the closely located SNP rs3740400 was differentially methylated depending on genotype; however, the methylation levels were very low and this SNP is probably not functional in determining the expression status of AS3MT. We previously reported that the SNP rs3740400 is associated with reduced gene expression of AS3MT [12], but these data indicate that, despite the fact that rs3740400 creates a CpG site, its effect is probably through linkage with other SNPs in the haplotype. It was unexpected that the AS3MT haplotype was associated with reduced AS3MT expression, as this haplotype is associated with more proficient arsenic metabolism and more dimethylated arsenic (DMA) in urine [12]. AS3MT is capable of methylating inorganic arsenic to methylarsonic acid (MMA) as well as MMA to DMA [9], [14]. Inorganic arsenic converted into MMA by AS3MT can be released as MMA, undergo a second round of methylation to DMA, or dissociate as MMA bound to glutathione [14]. Why does the low expression of AS3MT correlate with increased production of DMA whereas the high expression of AS3MT correlates with augmented synthesis of MMA? A possibility is that polymorphisms in other genes linked to the efficient AS3MT haplotype influence the methylation capacity. Clearly, this surprising finding needs to be analyzed further.
The strongest associations with AS3MT haplotype and DNA methylation were found for CpG sites related to CNNM2 (three CpG sites), which in turn were associated with the expression of AS3MT, CNNM2, NT5C2, and USMG5. CNNM2 is a magnesium transporter and widely expressed throughout the body. It is mutated in the rare disorder familial hypomagnesia [15] and CNNM2 SNPs were shown to influence magnesium concentrations in serum [16]. NT5C2 is an enzyme that dephosphorylates non-cyclic nucleoside monophosphates to produce nucleosides and inorganic phosphates [17] and its expression in muscles appears important for regulating energy metabolism and glucose transport [18]. Recently, SNPs in NT5C2 and the nearby gene CYP17A1 were found to be associated with fat mass in Japanese women [19]; these SNPs have also been associated with blood pressure [20], [21]. USMG5 was first recognized as a gene product whose mRNA level increased during skeletal muscle growth in rats [22], and more recently it was shown to be important in maintaining the ATP synthase population in mitochondria indicating that USMG5 is also important in cellular energy metabolism [23]. A recent genome-wide association study of the arsenic metabolism phenotype found SNPs near AS3MT to be associated with USMG5 expression [24].
Furthermore, our data showed that DNA methylation of three other genes (TRIM8, CALHM2, and C10orf26) was associated with the AS3MT haplotype. One report shows that TRIM8 is a direct target of p53 and that TRIM8 induces p53 stabilization and promotes the degradation of MDM2, which in turn directs the p53 response toward growth arrest and not apoptosis [25]. Under stress conditions, p53 promotes transcription of TRIM8. CALHM2 belongs to a family of transmembrane glycoproteins that has been suggested to modify calcium homeostasis [26], where three members are closely situated on chromosome 10 [25]. The function of C10orf26 is unknown. We did not find any association with other closely or more distantly situated genes to AS3MT, although several CpG sites per gene were present on the array. Due to the metabolism of arsenic through methylation by the same methyl donor as for DNA methylation, alterations of DNA methylation have been suggested as a toxic mechanism of arsenic [27]. Arsenic exposure per se appeared not to strongly modify the methylation or expression of this chromosome region. One speculation is that AS3MT also has other, arsenic-independent, functions in the body and factors related to such a function regulate AS3MT expression.
Some methodological aspects need to be commented upon. Firstly, four of the top methylation sites contained SNPs in and around the CpG site, which could bias the results due to impaired base extension since the probe cannot hybridize properly. The influence is stronger the closer the SNP is to the CpG site. Two SNPs (CNNM2 cg11667387 and USMG5 cg18367433) were situated less than 10 bp from the CpG site and this warrants caution. However, most of the top methylation sites (eight out of twelve sites), including the top site cg03493300, did not include any SNPs nearby the CpG site.
Secondly, haplotype was treated as a quantitative variable due to the fact that earlier studies of ours [12], [28], [29] have shown a strong allele-dose effect in these populations. The allele-dose effect is also clearly shown in the graphs presented in the supplemental material. In addition, analyses conducted using haplotype as a qualitative variable showed very similar results.
Thirdly, the efficient AS3MT haplotype is common in Argentina, but much rarer in Bangladesh. This lack of power in the Bangladeshi population may be a reason to the failure to detect some associations that were present in Argentina but not in Bangladesh.
Fourthly, we have measured DNA methylation in blood, which is a mixture of different cell types with different methylation pattern. We did not have the possibility to sort cells in the blood samples, as this was very difficult to achieve during the field studies. However, we knew from questionnaire data if the study participants had any chronic or acute disease at the time of sampling. These few individuals were excluded from the methylation analysis. Still, the consistency of results between the populations in terms of the associations with haplotype indicates that this likely isn't a concern in those analyses. We also measured gene expression in blood and not in liver, which is the main metabolizing organ for arsenic, and the DNA methylation and gene expression pattern of AS3MT may different in other tissues. Still, there most likely is a similar activity in blood based on the expression (albeit relative low) of the AS3MT gene in blood cells [30]. Obtaining tissue samples from individuals in screens of large cohorts is ethically problematic and extremely cumbersome, e.g. if liver biopsies are to be taken and preserved for analysis in the field of the Andes mountains, in contrast to blood sampling, which makes it the method of choice when applicable.
In the region around AS3MT, linkage disequilibrium (LD) exists, as noted in Argentina and Bangladesh [12], and the size of the LD block was recently characterized to be at least 350 kilobp in populations in Mexico [31]. A large LD block is present in Asian and European populations as well [10], [31]. One can speculate from the fact that the LD block is widespread in different populations, and that we here show a close relationship between haplotype, gene methylation status and to some extent also gene expression in an even larger region around AS3MT, that the LD block contains several genes important for a phenotype, possibly related to arsenic metabolism or to energy metabolism, as suggested from the functional role of several genes in the region.
Materials and Methods
Study Populations
Main cohort argentina.
Participants were women (N = 103) living on the Andean plateau in Northern Argentina and exposed to varying levels of arsenic from their drinking water [32], [33]. Most of the study participants (N = 71) were from the village San Antonio de los Cobres (water arsenic 200 µg/L), with about 5,000 inhabitants, while the remaining participants (N = 32) were from small surrounding villages (with various levels of water As, range 7–73 µg/L). The study individuals were recruited with the assistance of medical personnel, except in the small mining village Tolar Grande, where we went from house to house, explained the project and invited the adults to participate. We included only women, as men were often away from home for work for most of the day, and therefore drank water from different sources. The study area has minimal industrial or traffic pollution. The women in this society rarely drink alcohol or smoke tobacco. 94 women were included in the DNA methylation analyses and 90 women were included in the gene expression analyses, based on availability of high-quality DNA and RNA: 81 women were included in both analyses. Furthermore, 55 of the 103 women were previously analyzed for expression of one transcript in the AS3MT gene [12] and these data were included in the analysis of DNA methylation versus gene expression.
Bangladeshi cohort.
In short, our mother-child cohort in rural Bangladesh is nested in a large, randomized, population-based food and multi-micronutrient supplementation trial, which evaluated nutritional and environmental impacts on pregnancy outcomes and child health [34], [35], [36]. The cohort consists of a sample of 127 women delivering singleton infants at the central Matlab hospital or any of the four connected subcenters during early daytime.
Ethics Statement
Both verbal and written informed consents were provided by all adult study participants. In Bangladesh, mothers' gave verbal and written informed consent for use of their cord blood samples. In Argentina, all study subjects gave verbal and written consent, and informed verbal consent was obtained from the next of kin, caretakers, or guardians on the behalf of minors participants involved in the study (N = 4), while the minors participants themselves gave verbal and written consent. The process was documented in special consent forms. The studies, as well as the consent procedure, were approved by the Ministry of Health in Salta, Argentina, the Ethical Review Committee of ICDDR,B, Bangladesh and the Regional Ethical committee at Karolinska Institute, Sweden. The authors complied with all of the legal requirements pertaining to the locations in which the work was done. The data described in our publication are freely available upon request apart from information that will break the confidentiality rules for the study participants.
Blood and Urine Collection
Peripheral blood for DNA extraction was collected in K2EDTA tubes (Vacuette®, catalogue nr. sc-359548) and blood for RNA extraction was collected in PAX tubes (Beckton Dickinson, catalogue nr. 762165 Franklin Lakes, NJ). Spot urine samples were collected and processed as described previously [37].
Analysis of Arsenic in Blood and Urine
All analyses were performed in the same laboratory. Exposure to inorganic arsenic was assessed by the sum concentration (referred to as total arsenic) of inorganic arsenic, MMA and DMA in urine. Speciation of arsenic metabolites in urine (Argentina) was performed by high performance liquid chromatography (HPLC; Agilent 1100 series system, Agilent Technologies, Germany) coupled with hydride generation and inductively coupled plasma mass spectrometry (ICPMS) (Agilent 7500ce; Agilent Technologies, Japan), employing adequate quality controls [12]. Arsenic in urine from the Bangladeshi women was measured using hydride generation atomic absorption spectrophotometry. The total arsenic concentrations were adjusted to the mean specific gravity (SG) (1.020 g/mL for Argentina and 1.012 for Bangladesh).
DNA Isolation and Epigenetic Analysis
DNA was isolated using QIAamp® DNA Blood Midi kit (Qiagen, catalogue nr 51183). DNA quality was evaluated on a NanoDrop spectrophotometer (NanoDrop Products,Wilmington, DE) and a Bioanalyzer 2100 (Agilent, Santa Clara, CA) and showed good quality (260/280 nm >1.80). DNA was bisulfite-treated using EZ DNA Methylation kit (Zymo, catalogue nr D5001). The SCIBLU facility (Lund, Sweden) used 200 ng bisulfite-treated DNA for hybridization to the Infinium HumanMethylation 450K BeadChip (lllumina,catalogue nr WG-314-1003). The rs3740400 pyrosequencing assay was evaluated with individuals in Argentina (N = 94). The assay was designed using PyroMark Assay Design 2.0 (Qiagen) software encompassing rs3740400 allelic site with two additional CpG sites. The forward primer was biotinylated. PCR was performed using PyroMark PCR reagents (Qiagen, catalogue nr 972807). The PCR product was purified using Streptavidin Sepharose High Performance beads (Amersham Biosciences, catalogue nr 17-5113-01). Pyrosequencing was done using the PSQ HS96 Pyrosequencing System (Qiagen). The presence of SNPs in probe or query sites was evaluated according to http://www.rforge.net/IMA/snpsites.txt, which contains a list of SNPs in and around the CpG site.
The AS3MT haplotype analyzed is constituted by (5′ to 3′ direction) rs7085104 G, rs3740400 C, rs3740393 C, rs3740390 A, rs11191439 T, rs11191453 C, rs10748835 A, and rs1046778 C. All SNPs, but one (rs11191439), are non-coding. Haplotypes were inferred by PHASE [38]. Argentina: Genotyping for these eight SNPs in AS3MT was performed in blood DNA using Sequenom technology (Sequenom Inc.,San Diego, CA) [12]. Genotype data was missing for one individual. Bangladesh: rs7085104, r3740400, rs3740393, rs3740390 and rs1046778 were genotyped by Taqman allelic discrimination assay (ABI 7900,catalogue nrs. C___3284563_10, C__27510174_10, C__25804287_10, C__27510172_10, C___9596558_10). Genotype frequencies are presented in Table S2. Previous analyses of the LD pattern in another study group from Bangladesh showed that these five SNPs provide a good proxy of the full eight-SNP haplotype [12].
Gene Expression Analysis
RNA was extracted using the PAXgene Blood RNA kit (PreAnalytiX, catalogue nr 762174). RNA concentration and purity were evaluated on a NanoDrop spectrophotometer and RNA integrity (RIN) was evaluated on a Bioanalyzer 2100; the results showed good RNA quality (RIN>7.5). DirectHyb HumanHT-12 v4.0 (Illumina, catalogue nr. BD-103-0204) was used for gene expression analysis, at the SCIBLU facility of Lund University. Gene expression data were filtered from background signals with the BioArray Software Environment (BASE) software [39]. In this study, we only evaluated gene expression for the genes that demonstrated an association between AS3MT haplotype and DNA methylation pattern. Additionally, we used previously generated data from quantitative real time PCR analysis for expression of AS3MT (covering the 3′untranslated region (UTR) (Applied Biosystems Assays-by-design) as further described in Engstrom et al [12]. This was analyzed for 55 individuals; these subjects were matched for age, weight, body mass index (BMI), arsenic in urine, with a more narrow range of arsenic in urine than the individuals included in the HumanHT-12 v4.0 assay (Table 1). Relative gene expression for the AS3MT 3′UTR assay was evaluated according to individuals with 2 copies of AS3MT haplotype, for which the median gene expression was set to 1 [12].
Analysis of SNPs or DNA Methylation Sites in CpG Sites, Shores, Shelves and Islands or Enhancers
Whether the SNPs in the AS3MT haplotype introduced or removed a CpG site was evaluated by investigating the sequence around the SNP [40]. Whether the SNP or DNA methylation site was situated in a CpG shelf, shore or island was evaluated by using Emboss CpGPlot, which detects CpG-rich areas [40]. Enhancer elements for DNA methylation sites were defined according to annotations by Illumina (GPL-13534, available at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GPL13534).
Statistical Analysis
Methylation levels are specified by beta (β)-values that represent the fraction of methylation in the sample and range from 0 (unmethylated) to 1 (methylated). β-values were extracted from BeadStudio software (lllumina, San Diego, CA, USA). Missing values were imputed using k-nearest neighbour imputation (k = 10). 0.1% of the values were missing in both Argentina and Bangladesh. Potentially, missing value imputation may increase the correlation in the data. In our case, with so few missing values, this does not shift the data. Principal component analysis (PCA), which capture the major directions of variation in the data, was performed in order to evaluate the influence of technical and biological variables on DNA methylation. For the PCA, we employed the universally applicable singular value decomposition (SVD). The PCA showed that methylation levels were correlated with analysis plate in Bangladesh (two plates were used, while only one plate was used in Argentina). We removed this association from the data by the following procedure: for each methylation site a linear model was fit with the analysis plate as regressor. The residuals of the linear model become the new data for the methylation levels. In this way, the levels of each of the methylation sites were unrelated to analysis plate. No other variables had a major impact on general DNA methylation.
The associations of AS3MT haplotype with methylation levels were determined using Pearson correlation (haplotype was used as a numerical variable according to the number of copies, 0–2). Resulting p-values from the 450 K array were corrected for multiple testing (adjusting for 450,000 comparisons) by the Benjamini-Hochberg method to obtain false discovery rates (FDR). In this study we considered FDR <0.05 to indicate significant findings.
The top methylation sites were then further evaluated by building multivariable models. First a univariable model was performed, with each methylation site used separately as a dependent variable, and number of copies of AS3MT haplotype was employed as an independent continuous variable. We also evaluated a model using number of copies of AS3MT haplotype as a categorical variable; however the results were very similar, indicating an allele-dose effect, and thus haplotype was used as a continuous variable. Multivariable models were built by evaluating, in univariable models, the association between each methylation site and other variables potentially influencing DNA methylation. The variables evaluated in Argentina were urinary arsenic, age (years), BMI, and use of coca leaves, while the variables evaluated in Bangladesh were arsenic measured in maternal urine around gestational weeks 6–8 or 30, birth weight, mothers' age (years), mothers' BMI, asset score (similar to socioeconomic status), sex of the baby and gestational age. Variables significantly associated with a methylation site were included in the multivariable models (Table 3). We also evaluated if the haplotype groups were significantly different regarding the potentially influential variables, in order to evaluate stratification according to haplotype (Table 1). This was evaluated with the non-parametric Kruskal-Wallis test.
The methylation sites that demonstrated a positive association with AS3MT haplotype were further analyzed for association with gene expression as well as with total arsenic in urine. The association between AS3MT haplotype and gene expression was also. These analyses were evaluated by Spearman's correlation. Furthermore, a linear regression analysis was performed with gene expression as dependent variable, and methylation level as independent variable, where adjustments were made for total arsenic in urine and RIN. Thereafter, a linear regression analysis was performed, with gene expression as dependent variable, and copies of AS3MT haplotype, total arsenic in urine and RIN (to adjust for variations in RNA quality) as independent variables. The statistical analyses were completed by the use of SPSS 18.0 (SPSS Inc, Chicago, IL USA) and R version 2.14.2. Statistical significance refers to P<0.05 (two-tailed).
Supporting Information
Figure S1.
Influence of AS3MT rs3740400 on DNA methylation. Genotypes CA and CC are presented, since the genotype AA does not carry any potential CpG site.
Figure S2.
Influence of AS3MT haplotype on DNA methylation of AS3MT , CNNM2 and NT5C2 .
Figure S3.
Correlation in DNA methylation between genes around AS3MT compared to other regions on chromosome 10. This was done by analyzing windows of 20 consecutive genes and the fraction of CpGs that are correlated between the genes with Pearson correlation >±0.5 (excluding CpGs within the same gene). The figure shows a permuted (randomized) version of chr10 (green line in plot) with AS3MT indicated (dashed red line).
Figure S4.
The relationship between USMG5 DNA methylation and USMG5 gene expression, stratified for AS3MT haplotype.
Table S1.
SNPs and alleles included in the AS3MT haplotype studied.
Table S2.
Correlations (Spearman's rho coefficient) of DNA methylation in genes (5′-3′) in chromosome 10q24 associated with the AS3MT haplotype for A) Argentina and B) Bangladesh.
Table S3.
Overview of the gene expression levels for the genes associated with the AS3MT haplotype in Argentina.
Table S4.
Correlations in gene expression between genes in chromosome 10q24 (Argentina).
Table S5.
Correlations (Spearman ranks) between urinary arsenic (As), DNA methylation (horizontally) and gene expression (vertically) for TRIM8, C10orf26, CALHM2 , and USMG5 (all CpG sites were negatively associated with the AS3MT haplotype).
The authors thank Alejandro Cardozo, Plácida Mamaní, Esperanza Casimiro, and Eva Assarsson for recruiting the volunteers, Karin Paulsson for genotyping and Barbro Nermell, Margaretha Grandér and Brita Palm for measuring the arsenic metabolites in urine.
Author Contributions
Conceived and designed the research: KSE KB. Performed the research: MBH SA. Contributed to the study material: MV RR. Contributed to the manuscript writing: KSE KB ML SA MBH MV RR. Analyzed the data: ML KSE.
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Online piracy
ABA adds its voice to concerns over online piracy, counterfeiting
The American Bar Association has called for "widespread adoption and expansion of voluntary measures" among businesses using the Internet, to "discourage or eliminate online piracy and counterfeiting."
Reviewing 5 of June's top stories
News Corp.'s restructuring, HP's legal battles, a Megaupload extradition controversy and more
IP: Is ACTA Europe's SOPA?
On Jan. 18, 2012, neglected encyclopaedias were taken down from bookshelves, as Wikipedia.com closed its virtual doors to people in the English-speaking world. They did so in protest against the Stop Online Piracy Act (SOPA) and the Protect Intellectual Property Act (PIPA), two pieces of prospective U.S. legislation regarding the...
Canadian legislature to rule on controversial copyright bill
As the furor over copyright legislation dies down in the U.S., the controversy is heating up north of the border, where the Canadian legislature is debating the controversial Bill C-11.
Public advocacy group proposes 5 alternatives to SOPA
Copyright infringement is a hot-button issue, especially in today's age of digital piracy and illegal file sharing. Efforts to combat online theft in the form of the Stop Online Piracy Act (SOPA) failed earlier this year. But now, a public advocacy group is offering its own plan to balance Internet...
IP: Is SOPA really that bad?
Two weeks ago we experienced a moment of Internet history. On Jan. 18, dubbed "Blackout Day," Internet giants such as Google, Reddit and Wikipedia acted in protest in an attempt to show how overreaching the Stop Online Piracy Act (SOPA) and the Protect IP Act (PIPA) are by censoring themselves...
Wikipedia, Reddit and others shut down to protest SOPA and PIPA
Today, websites across the Internet are tapping into their Twisted Sister personas, telling Congress that, when it comes to proposed online piracy legislation, they're not gonna take it.
Obama administration says it won't support SOPA
Technology companies and Internet businesses breathed a sigh of relief on Saturday when the White House issued a statement saying it wouldn't support Congress' controversial proposed online piracy legislation.


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We often see accomplished singers and musicians and marvel at their gifting. I personally am a big fan of Israel Houghton. In case you don't know, he is a contemporary gospel singer. He writes most of his own music, plays guitar, and sings very well. He is talented beyond words and has a sound that crosses cultural boundaries.
The Fruit of Investing Your Time
Thanks for stopping by my blog! I enjoyed reading yours. Out of total curiosity, where did you find that top 100 list, I had no clue!
Glad to read that you got your desire.
Thanks to everyone for your encouraging comments! Speech delays are more common than I thought, especially among boys!
My son receives free speech therapy through a program in our state. His speech therapist sees him once every other week. She used to come every week but he progressed.
@ Krystal – My son has a ministry calling too. I've been seeing some of the recipes you have been sharing. They look great and one day, I'm going to try one of them.
@ Jolyn – people were telling me that I didn't need to put my son in speech too, but I also had a gut instinct. He's doing well now, but I think the speech therapy has made a huge difference. Plus, it has taught me different ways to help my son communicate that I was not aware of before.
@ Heidi – I'm going to send you an email. Thanks for stopping by!
PS, @ChristianMommyWriter …
Are you based in Fayetteville?
We lived there 7 years! My husband went to Iraq twice and Afghanistan once. Would love to correspond with you directly about your experiences and to pray with you if your husband is deployed … Also, if you haven't checked out PWOC at Fort Bragg yet, I highly recommend it!
🙂 My email is email@example.com.
That's so cool, Jolyn.
I should probably look into speech therapy for my little guy because of his struggle with the rs and ls and ws.
What is interesting is that when he tries to spell words now, he will pronounce an "r" like a "w" but if the word calls for an "r," he will write the "r." He knows what he's saying but just can't get his tongue around it. I'm glad you posted that comment about your son's therapy. It has started me thinking!
I am so happy for you! What a wonderful milestone.
We had so many well-meaning people telling us not to worry about our son's speech. My instincts told me otherwise, and he ended up being diagnosed with a speech disorder and qualifying for some amazing therapy. He is four now and they all say his progress is unprecedented! God is good.
I wanted to listen to all of those friends and family who told me he "just needed more time" and "boys just don't talk early but one day it will just all come out", etcetc. I am glad I followed my instincts (which I believe can come from prayer and the Holy Spirit) that told me he needed more than "time".
Keep following your instincts! I have now witnessed countless hours of professional therapists working with children, and I am amazed at how much of speech therapy simply consists of talkingtalkingtalking ad naseum. One-on-one, on the floor (or at the table), talkingtalkingtalking, and engaging them non-stop. Keep it up!
I LOVE THIS POST! And I love your blog. Beautiful.
I visited after you kindly posted on my blog. Thanks so much. 🙂
On your son's verbal abilities, don't worry, seriously. My little boy is now 5, and he also was woefully behind everyone else in speech. He struggles currently with rs, ls and ws, but other than that is now doing great — in fact, we can't get him to stop talking! Ha ha!
Another thing that helped is that ever since he has been a newborn, I have done story time with him every single day. Currently, we go to our public library once a week and spend about an hour to an hour-and-a-half just picking out books. They allow us to check out 25 books, 10 VCR tapes and 5 DVDs at one time! I literally take a large apple picking basket every week.
It's so much fun to come home with our "treasures" and start a week with new books. We average 45 minutes per night for story time in bed now, every night, and my little boy is now reading at the level of a mid-school-year first-grader.
I really wasn't reading to him to get him to perform — just to introduce my love of reading to him and to get him loving it, too. People are amazed at how he reads now!
You seem like an awesome mom. I'm sure your little boy will excel when it's time.
And hey, guess what else — Albert Einstein did not speak one word until the age of 5. So there ya go.
Have a great day, and God bless!
Heidi Rafferty
Harrodsburg, KY
My son had delayed speech, too. So he qualified for speech and occupational therapy. Both therapists came out to our house once a week. The program ran through age 3. Then at age 3, he qualified for the Early Childhood program at our local elementary school. And since he started that program, with other children, his speech has exploded!
To the point that I have to tell him sometimes, "Adam, no talking."
But you know, the Lord told me years ago what Adam's calling was in life. And it involved speaking in the ministry. So I always knew that the speech delay was just a season. And now we're coming out.
Also, this post resonates with me because the Lord's been dealing with me also to spend more time with my children. I typically cook all my meals from scratch (which can take hours some days), do laundry, vacuum daily, blog, etc, etc. So it's VERY easy to just kinda turn on the TV and let the kids have the run of the house. But in my opinion, parenting should be ACTIVE and HANDS-ON, not passive.
This is part of the reason why I've not been posting on my own blog as much lately. I don't want to spend an hour on the computer that could have/should have been devoted to the kids.
Good post. I think we all have those balance issues that you talked about. But the Lord straightens us out, doesn't He? 😉
Next post: Dysfunctional Diet
Previous post: Walking for a Cure!


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We often write about businesses and entrepreneurs who have chosen bitcoin as an instrument of payment process development. At the moment bitcoin is likely one of the most creative payment method for most countries and businesses. But Project Bitcoin use cryptocurrensy in a truly creative way.
Let me assure you that the whole idea of Project Bitcoin is worth to pay attention to. It is said that "The vision of Project Bitcoin is simple: turn a special central bank issued (bit)coin into a house by a lake by making 20 increasingly larger international transactions using only bitcoin and the people who believe in it. " To do it, they "must sell that coin for more than its value, and then make buy/sell bitcoin transactions that continuously increase in value until, hopefully.... the house".
The way it began
The project was launched by one of bitcoin activists - code name Bitcoincito, on 11th of January. On the page about the project Bitcoincito writes: "I became attracted to bitcoin because of its promise. It's promise, to me, is to liberate a world of its dependence on insane monetary policies decided on by politicians and bankers who hold all the chips and want to keep them. Despite my own heavy workload (I love what I do... even if it pays peanuts) I wanted to engage in an adventure, a dance, a love affair with bitcoin, and with people who believe in its potential. It was with this intention that Project Bitcoin was born".
Soon after the project was launched it received a great reaction from bitcoin users, manufacturers and artists.
The first transaction the project got was the Peruvian Central Bank issued 1 SOL (~$0.35) transformed into a new coin. Bitcoincito said: "I found a metalworker in Peru who was able to shave down one side of the coin and etch the bitcoin logo into it". So simple idea - turned to be quite successful.
A Plan for the 9th Transaction
Now Project Bitcoin represents a new Last (Bitcoin) Supper. It is the 9th transaction of the project.
Bitcoincito tells the story of Last Bitcoin Supper on the projects block page:
"About a month ago, I received an email from a French painter named Youl. A friend had introduced him to Project Bitcoin, and he felt called to be a part of it. I checked out his work, and I was floored. His work is a rare combination of heart, edge, and technical brilliance".
Soon after Youl and Bitcoincito began planning out new painting for the next transaction.
"After days of discussion, we decided Youl would paint a huge 140x70 centimeter canvas interpretation of Da Vinci's Last Supper. We believed that Youl's painting could tell the story of bitcoin with a profundity few words could even approach".
When the 9th transaction of Project Bitcoin will be done it could be considered as a great pay for truly creative work of art.
Last (Bitcoin) Supper
Both Youl and Bitcoincito spend number of nights in France learning about and understanding the history of Da Vinci and his painting. And especially – the stories of each of the disciples on the Last Supper, so that also each disciple of the new painting mapped to a person or archetype in the bitcoin community.
Also Youl added QR code on the painting that links to a wallet they set up that will donate 100% of any funds received in it to nonprofits specifically doing work to support bitcoin.
In the end of presentation Bitcoincito asked his readers and followers "If you buy this painting, please let me visit you and the painting sometime. Its the coolest piece of art I've ever seen, its meaning goes deep, and I feel such crazy gratitude for having the opportunity to play a small role in its creation!"
We wish a good luck to Bitcoincito and Project Bitcoin.
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Showgirl Classics Vanessa Del Rio's Fantasies
Showgirls Classics presents for your enjoyment the most famous and most beautiful women on the X-Rated screen. Filmed in more erotic and outrageous sexual acts than you have ever seen performed before. The finest direction, techniques and equipment were used to bring you the highest quality video tapes possible of these superstars of sex. "Vanessa Del Rio's Fantasies "Superstar Vanessa Del Rio gives you thrill after thrill as you watch her live out all of her insatiable erotic fantasys. She enjoys a rendezvous with horny nurse played by Leslie Bovee, who's always been hot for Vanessa. After they stroke and suck each other into uncontrolled ecstasy, they find a patient in the waiting room and give him the surprise of his life. Then Sexy Samantha Fox brings two studs over for some searing four-way action. Later she joins breath-taking Crystal Dawn in a magnificent marble bath tub for some deeply orgasmic all-***** soaking wet action, And when Crystal brings out her special toys, Vanessa shows her just where she likes it best. She loves dildos and proves it later when she joins nasty June while she rides her boyfriends huge manhood. Vanessa and her dildo find some interesting ways of getting June to cum even more. And for a grand finale, she shows her tongue teasing skills to her neighborhood mechanic, who is a very willing subject- so much so that he shows her his own special talent. All in all, this super collection of Vanessa Del Rio at her wickedest is a true collector's 'must have'! Enjoy.
- Duration:
- 15:00
- Studio:
- Year:
- 2008
- Added:
- 8 years ago
- Views:
- 2199
- Comments:
- 0
- Price:
- $ 1.02 (For 24 hour streaming views)
- Performers:
- Leslie Bovee
- Crystal Dawn
- Vanessa Del Rio
- Samantha Fox
- Tags:
- fetish
- hardcore
- anal
- Categories:
- Fetish


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More Divorcing Israeli Spouses Seek Straw Men to Conceal Holdings
It is increasingly common for wealthy individuals to conceal their wealth to avoid having to split it with the ex-wife or ex-husband.
Yaakov Birnbaum was one of several people convicted earlier this month of concealing shareholdings in a company active in Romania. But Birnbaum's reason for turning to Pacifica Holdings from 2007 to 2008 was different than that of the other defendants. His principal motivation was "his desire to hide his holdings from his wife," the Tel Aviv District Court found.
In recent years it's become increasingly common for wealthy individuals to seek out straw men to whom they can sign over their assets in order to conceal their true wealth from the spouses they are in the process of divorcing. The goal is to avoid having to split this portion of the money with the ex-wife or ex-husband.
Many wealthy people, said attorney Maya Weisman, "feel it's not fair that half should go to their husband or wife, who didn't do anything to earn it. They're also thinking about the future. And of course, there's the desire for revenge."
Assigning assets to someone else has become a popular tactic because it is "the least expensive and least complicated" method of hiding assets, added attorney Gil Dachoach.
Nevertheless, this system entails a not insignificant risk: The person to whom you sign over your assets may decide not to give them back.
That, according to Birnbaum's testimony to the Israel Securities Authority, is what happened to him. In a bid to conceal money from his wife, he transferred "a few million" to a special bank account that a friend set up for the purpose. The account was in his friend's name, but in theory, the money was his for the asking. In practice, he said, his friend refused to give it back.
This risk causes many wealthy people to prefer a more complex method of hiding assets: opening secret accounts in tax havens overseas.
Weisman, who originally worked in financial law but then switched to family law, said that often "a woman will come to me who doesn't know the first thing about how much property her husband really has." Consequently, she said, one the first things Weisman does is look into that question.
A few months ago, for instance, "a woman walked into my office, a lawyer from a respected firm, and told me she and her husband had decided to divorce amicably, by agreement." After all, the woman said, all they have is their house and a few savings, so "we don't have anything to fight about, right?"
But when Weisman started questioning her, it turned out her husband had once owned a company. The woman thought it was defunct - but with a little research, Weisman found out that it still existed, under another name. It was owned by another Israeli company, which in turn was owned by another company. After following the chain of ownership back through another few companies, Weisman discovered that the ultimate owner was a company registered in Cyprus - owned by the woman's husband.
At that point, the woman tore up the draft divorce agreement she had been ready to sign and went to court instead.
In particularly complex cases, Weisman said, she recommends that clients hire a private investigator to look for their spouses' assets.
Attorney Yael Novogrodsky said that while holdings in Israel are relatively easy to locate, private investigators are essential in locating overseas assets. "If a suspicion arises that money has been smuggled abroad, then even before getting started in court, you ask a private investigator to do the preliminary spadework," she said.
Nevertheless, family courts do have broad powers to help divorcing couples unearth their partners' hidden assets. These courts readily issue discovery orders and frequently even impound assets to enforce them. Often, they also bring in accountants to assist the discovery process.
David Goldberg, deputy chairman of the Institute of Certified Public Accountants in Israel, said that in such cases, he goes through all the available financial records looking for "unreasonable data" that "sets off a red light."
An example of that? "If two weeks before your examination [of the records], or close to the time of the divorce, you suddenly see an asset that has been transferred to an unrelated party at an exaggerated price," says Goldberg.
By law, each spouse's personal computer is off limits during this process. But a family computer is fair game, and courts will often issue a writ allowing lawyers or private investigators to seize a family computer and search the files for information about hidden assets.
Novogrodsky noted that the process can be extremely unpleasant for the partner suspected of concealing assets, ranging from intensive scrutiny of all his known financial records to having assets impounded or even being subject to writs restricting his activities.
"It's a mess that you're better off not getting into," she concluded. "It's better to just sign property agreements."
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In MySQL, I can use the replace statement which either updates the data there or inserts it. Is there a comporable syntax to use in postgreSQL? I need to do an insert and don't want to have to worry about if the data is already there or not...so don't want to see if it there, if so do update if not insert...etc. Thanks.
Next: From: Uzo Madujibeya Date: 2005-10-29 01:57:45 Subject: Backup problem...No BIN directory!!! Previous: From: Robert Treat Date: 2005-10-29 01:47:17 Subject: postgis / mediawiki / postgresql was Re: [Freegis-list] Re: [GENERAL] Map of Postgresql Users (OT)


text: Its time to talk about depression. With a cancer diagnosis causing such impact and distress to the lives and families of the recipient, it can be sadly all too common for feelings of depression to accompany cancer patients. The European Cancer Organisation (ECCO), the voice of multidisciplinarity in cancer, recognises the important needs of cancer patients in respect to mental health and is therefore pleased to support the World Health Organisation's awareness drive on depression on the occasion of World Health Day, 7th April. ECCO President Peter Naredi remarked: "The focus of World Health Day on depression is an excellent reminder to all of us to challenge mental health stigmas wherever we find them, and create an environment around us where all can feel open and encouraged to discuss their feelings. One of the honours of leading an organisation such as ECCO is being in such regular contact with colleague professionals in psycho-oncology, a crucial part of the multi-disciplinary cancer team. In a shortly-to-be-published position paper on innovation, ECCO will emphasise the need for cancer patients to have access to psycho-oncology services. This ensures that not only the direct medical needs of cancer treatment are provided for, but also patients' psycho-social support requirements. We all need to keep talking about depression and mental health to ensure we create the architecture for treatment that patients have a right to expect." About World Health Day The founding of the World Health Organisation (WHO) is celebrated annually on April 7th. Each year, this anniversary allows for the highlighting of a specific health topic and provides a platform for global mobilisation through World Health Day. The 2017 campaign theme is depression. With a campaign slogan of "Depression: Let's talk", WHO seeks to bring depression into the spotlight of international medical dialogue. From 1990-2013, individuals suffering from depression and/or anxiety increased by close to 50%. Depression does not discriminate, causes mental stress and impedes an individual's ability to carry out simple tasks. At times, depression also leads to the severing of important relationships; however, it can be treated. Understanding depression and how it can be prevented and treated will encourage greater discourse which can remove the stigma associated with it, encouraging more people to seek help. About ECCO ECCO is a not-for-profit federation comprised of 25 Member Societies cumulatively representing over 170 000 medical professionals. It is the only multidisciplinary organisation that connects and responds to all relevant oncology stakeholders throughout Europe. ECCO aims to promote and uphold the rights of all European cancer patients with regards to receiving the best treatment and care, while promoting dialogue amongst all organisations involved in cancer at a European level. Page last modified: 20 April 2017


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Chemical Rush
Credits & Info
- Uploaded
- Dec 13, 2008 | 7:30 PM EST
- File Info
- Song
- 2 min 55 sec
- Score
- 4.44 / 5.00
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Please contact me if you would like to use this in a project. We can discuss the details.
- Score:
- Rated 4.44 / 5 stars
- Plays & Downloads:
- 21,164 Plays | 1,429 Downloads
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Author Comments
Rated 5 / 5 stars
My ears.....they gasm
I happened upon this little gem while parsing the "Top Rated of All Time" list on the front page. I must say, my friend, I can understand why this is on that list. You kept it true to the original, but added your own personal touch. This is what I expect out of a remix of a song. You didn't muddle through with a million layers of synth or bass. You DnB'd it up a little, which my inner producer applauded for. The quality of the track is great, as well. No overshoots or major bangs out of whack. This is ready to be a master track. Congrats. You've earned this rating. I'm looking forward to listening to some more of your work.
Wow, thanks so much! Haha, I'm flattered.
I'm glad you enjoyed it!
Thanks for your review, and thanks for listening! c:
Rated 5 / 5 stars
Beautiful Ear-gasim.
It sends shivers down my spine when i hear it. Beautiful.
Rated 4.5 / 5 stars
A New Meaning to Catchyness!
The sonic Loop is workin for ya.
Not bad,Not bad at all.
Rated 5 / 5 stars
Sounds fantastic..I really love the pace and you didn't over do it. Its so easy when remixing a already good sounding song to just throw lots of instruments in, but you disclipined yourself it seems. Thanks!
Rated 5 / 5 stars
OMG Bring back so many memories!!!.
Best level on sonic. First played it 13 years ago. remember it like it was yesterday.


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The new king of the road extends the Indian Motorcycle lineup debuting at Sturgis.
Indian Motorcycle®, America's first motorcycle company, is pleased to announce the all-new Indian Roadmaster®, setting a new benchmark as the ultimate in luxury motorcycle touring. This exceptional new addition to the Indian Motorcycle family will make its public debut on August 2nd at the 74th annual Sturgis Motorcycle Rally. More announcements regarding the 2015 line of Indian bikes will also be made on August 2nd.
All-New Indian Roadmaster
Originally produced from 1947-1953, the Indian Roadmaster was widely recognized as one of America's most reliable and comfortable touring bikes, and the all-new Roadmaster is destined to reclaim that honour. In keeping with its legendary namesake, the Roadmaster comes loaded with premium features for comfort, convenience and luxury, including expansive storage capacity and unmatched power and performance.
"It has truly been a privilege and an honor for our team to be a part of motorcycling history in designing and reintroducing the Roadmaster, which has such a legendary reputation as the king of luxury touring bikes," said Steve Menneto, VP of Motorcycles for Polaris Industries. "The quality, craftsmanship, performance and standard feature set on this bike is simply unmatched in the luxury touring segment, and we look forward to sharing the all-new Roadmaster and other exciting announcements with the global motorcycling community beginning August 2nd in Sturgis."
Comfort & Convenience by Design
The new Roadmaster was built from ground up for all-day, two-passenger touring comfort, with expansive, easy-access storage for extended rides in virtually any weather condition. If offers:
Luxury is Standard
The all-new Roadmaster takes American motorcycle luxury and comfort to a whole new level with an extensive list of standard premium features that include:
Proven Power and Endless Chrome
Indian Motorcycle's new flagship luxury touring bike is powered by the awe-inspiring and proven Thunder Stroke 111™ engine, a 49-degree, air-cooled V-twin power plant mated to a 6-speed overdrive transmission. The Roadmaster is dripping in standard chrome, including the forks, trim, top box rack, front and rear highway bars, fender tips, and many other chrome components that are options on other brands.
Pricing & Availability
The all-new Indian Roadmaster is available from $38,995 Ride Away with expected arrival in Australia in late October early November. It is available in Thunder Black, Indian Motorcycle Red, and a two-tone Indian Motorcycle Red/ Ivory Cream paint scheme.
Visit www.indianmotorcycle.com.au to locate an Indian Motorcycle dealer near you, and to see the full line of apparel, parts and accessories.
Like Us at facebook.com/indianmotorcycleaustraliaandnewzealand
Indian Motorcycle joins Victory Motorcycles (founded in 1998), as a wholly-owned division of Polaris Industries Inc. (NYSE: PII). Indian is America's first motorcycle company founded in 1901, winning the hearts of motorcyclists around the world and earning distinction as one of America's most legendary and iconic brands through unrivaled racing dominance, engineering prowess and countless innovations and industry firsts. Today that heritage and passion is reignited under new brand stewardship. Polaris Industries, Inc. is proud to own and manufacture the oldest and newest American motorcycle brands. To learn more, please visit www.indianmotorcycle.com.au.
Always wear a helmet. Never drink and ride. © 2016 Indian Motorcycle International, LLC
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text:
Municipal and State Lawyers and Law Firms in Uruguay
Municipal and State Law Lawyers in Uruguay
Blanco & Etcheverry - AbogadosMontevideo, Uruguay
Full-Service Legal Firm(598)2 915-5548
Fernandez Secco & AsociadosMontevideo, Uruguay
Montevideo, Uruguay Intellectual Property and Corporate Law Attorneys+598 2 9161913
Since 1980 our firm has become a multi disciplinary team of Attorneys, Accountants and Public Notaries. ; At Fernandez Secco & Asociados we provide attentive and proactive assistance to our clients by delivering value-added solutions. We provide services of the highest quality due to our...
Fraschini & AsociadosMontevideo, Uruguay
Full-Service Legal Firm in Uruguay598 (2) 900 69 68
Galante & MartinsMontevideo, Uruguay
Full Service Law Firm in Uruguay+598 2 708 7172
Tradition: More than half century of tradition they consolidate to Galante & Martins like referring between the companies of professional services. Since 1951, the company/signature has successfully developed areas of agreed services to the present exigencies of the enterprise world and of...
Navarro AbogadosMontevideo, Uruguay
Full-Service Law Firm in Uruguay+598 2 915 0266
VDM ConsultoresMontevideo, Uruguay
Foreign Investment, Tax, Labour, Banking and Partnership Law Firm in Montevideo, Uruguay+598 2 9084793


text:
Responding to Requests: Why Reliability is the First Piece of the ROCC of Trust
Written By TotalTrust
We get lots of requests from people we know, students, alumni, clients, friends, family. We can't obviously say yes to all of these requests, but we do our absolute best to respond either yes or no, and not leave people wondering if we ever heard their requests. We actually try to respond within 24 hours, […]


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Yes, fruit flies speak up! They want to tell you that they, and their ancestors for nearly a century, have given their health and very lives to the cause of disproving evolutionary theory. The facts are in. Evolution is a fake. The fruit flies tell you so. This is science vs. evolution—a Creation-Evolution Encyclopedia, brought to you by Creation Science Facts.
CONTENT: Fruit Flies Speak Up
The Great Mutation Research Project: Nearly a century in progress
Speeding Up the Evolutionary Process: Their generations have been studied longer than the presumed time man has been on earth
A New Species Is Never Produced: The fruit flies always remain fruit flies
What Happens to the Mutated Fruit Flies?: We have had opportunity to clearly learn what mutations do to a physical organism
Summary: Once a fruit fly, always a fruit fly
This material is excerpted from the book, MUTATIONS. An asterisk ( * ) by a name indicates that person is not known to be a creationist. Of over 4,000 quotations in the books this Encyclopedia is based on, only 164 statements are by creationists. You will have a better understanding of the following statements by scientists if you will also read the web page, Mutations.
Well, they may not say much, but at least those who have spent years observing them surely have something to say. The humble fruit fly will speak to us through the researchers who have spent countless hours studying the varied ways in which mutations have damaged those flies:
Nearly a century in progress.
In 1904, Walter S. Sutton, an American cytologist, decided there might be some connection between Gregor Mendel's 1860s research and the newly discovered chromosomes with their genes. A major breakthrough came in 1906, when Thomas Hunt Morgan, a Columbia University zoologist, conceived the idea of using fruit flies (Drosophila melanogaster) for genetic research. This was due to the fact that they breed so very rapidly, require little food, have scores of easily observed characteristics and only a few chromosomes per cell.
"The fly could be bred by the thousands in milk bottles. It cost nothing but a few bananas to feed all the experimental animals; their entire life cycle lasts 10 days and they have only four chromosomes."—*R. Milner, Encyclopedia of Evolution (1990), p. 169.
Later still, fruit flies began to be used in mutational research. What that research revealed—settled the question for all time as to whether evolution could successfully result from mutations. And those little creatures should be able to settle the matter, for it takes only days for a fruit fly to reach maturity; after that it steadily reproduces young. Each of its offspring matures in a few days, and the generations multiply rapidly. What it would take mammals tens of thousands of years to accomplish, the humble fruit flies can do within a very short time.
We have heard about "the rocks crying out" (Luke 19:40). The fossil rocks surely are. Well, the little fruit flies have a testimony to give also.
Fruit fly generations have been studied longer than the presumed time man has been on earth.
According to evolution, man has lived on the earth for a little over a million years. Yet experiments on fruit flies have already exceeded the equivalent of a million years of people living on earth. Here is a clear statement of the problem: "The fruit fly has long been the favorite object of mutational experiments because of its fast gestation period [twelve days]. X rays have been used to increase the mutation rate in the fruit fly by 15,000 percent. All in all, scientists have been able to "catalyze the fruit fly evolutionary process, such that what has been seen to occur in Drosophila is the equivalent of the many millions of years of normal mutations and evolution."
"Even with this tremendous speedup of mutations, scientists have not been able to come up with anything other than another fruit fly. Most important, what all these experiments demonstrate is that the fruit fly can vary within certain upper and lower limits but will never go beyond them. For example, Ernst Mayr reported on two experiments performed on the fruit fly back in 1948.
"In the first experiment, the fly was selected for a decrease in bristles and, in the second experiment, for an increase in bristles. Starting with a parent stock averaging 36 bristles, it is possible after thirty generations to lower the average to 25 bristles, "but then the line became sterile and died out." In the second experiment, the average number of bristles were increased from 36 to 56; then sterility set in. Mayr concluded with the following observation: Obviously any drastic improvement under selection must seriously deplete the store of genetic variability . . The most frequent correlated response of one-sided selection is a drop in general fitness. This plagues virtually every breeding experiment."—*Jeremy Rifkin, Algeny (1983), p. 134.
The fruit flies always remain fruit flies.
After decades of study, without immediately killing or sterilizing them, 400 different mutational features have been identified in fruit flies. But none of these changes the fruit fly to a different species.
"Out of 400 mutations that have been provided by Drosophila melanogaster, there is not one that can be called a new species. It does not seem, therefore, that the central problem of evolution can be solved by mutations."—*Maurice Caullery, Genetics and Heredity (1964), p. 119.
*"Richard Goldschmidt fell into despair. The changes, he lamented, were so hopelessly micro [insignificant] that if a thousand mutations were combined in one specimen, there would still be no new species."—Norman Macbeth, Darwin Retried (1971), p. 33.
A thousand known fruit-fly mutations placed in one individual—would still not produce a new species!
"In the best-known organisms, like Drosophila, innumerable mutants are known. If we were able to combine a thousand or more of such mutants in a single individual, this still would have no resemblance whatsoever to any type known as a [new] species in nature."—*Richard B. Goldschmidt, "Evolution, As Viewed by One Geneticist," American Scientist, January 1952, p. 94.
The obstinate, stubborn little creatures!
"Fruit flies refuse to become anything but fruit flies under any circumstances yet devised."—*Francis Hitching, The Neck of the Giraffe: Where Darwin Went Wrong (1982), p. 61.
We have had opportunity to clearly learn what mutations do to a physical organism.
Fruit flies which receive mutations are always weakened in one way or another.
"The clear-cut mutants of Drosophila, with which so much of the classical research in genetics were done, are almost without exception inferior to wild-type flies in viability, fertility, longevity."—*Theodosius Dobzhansky, Heredity and the Nature of Man (1964), p. 126.
The mutated creatures die out, when placed out in nature with normal hardy specimens.
"A review of known facts about their ability to survive has led to no other conclusion than that they [the mutated offspring] are always constitutionally weaker than their parent form or species, and in a population with free competition they are eliminated . . Therefore they are never found in nature (e.g. not a single one of the several hundred [types] of Drosophila mutation), and therefore, they are able to appear only in the favorable environment of the experimental field or laboratory."—*H. Nilsson, Synthetische Artbildng (1957), p. 1186.
The classical example of the damaging effects of mutations is to be found in what scientists have done to fruit flies by inducing mutations in them.
"Most mutants which arise in any organism are more or less disadvantageous to their possessors. The classical mutants obtained in Drosophila usually show deterioration, breakdown, or disappearance of some organs. Mutants are known which diminish the quantity or destroy the pigment in the eyes, and in the body reduce the wings, eyes, bristles, legs. Many mutants are, in fact lethal to their possessors. Mutants which equal the normal fly in vigor are a minority, and mutants that would make a major improvement of the normal organization in the normal environments are unknown."—*Theodosius Dobzhansky, Evolution, Genetics, and Man (1955), p. 105.
No new-species fruit flies have ever resulted from sixty years of irradiation the poor creatures.
"It is a striking, but not much mentioned fact that, though geneticists have been breeding fruit flies for sixty years or more in labs all round the world—flies which produce a new generation every eleven days—they have never yet seen the emergence of a new species or even a new enzyme."—*Gordon R. Taylor, The Great Evolution Mystery (1983), p. 48.
Pitman says the experiments have only produced geneticists' monsters.
"Take the example of fruit flies (Drosophila). Morgan, Goldschmidt, Muller, and other geneticists have subjected generations of fruit flies to extreme conditions of heat, cold, light, dark, and treatment by chemicals and radiation. All sorts of mutations, practically all trivial or positively deleterious, have been produced. Man-made evolution? Not really: Few of the geneticists' monsters could have survived outside the bottles they were bred in. In practice mutants die, are sterile, or tend to revert to the wild type."—*Michael Pitman, Adam and Evolution (1984), p. 70.
Once a fruit fly, always a fruit fly.
The following news article sums it all up. Notice the fact that, in those instances in which damaged fruit flies survive long enough, they change back into regular fruit flies—even those without eyes!
"For 80 years scientists have been experimenting with the lowly fruit fly (Drosophila), trying to prove that all life on planet earth is the result of a series of `good accidents.'
"Evolutionists, through a marvelous leap of faith, believe that the almost endless variety and complexity of plants and animals `evolved' from an ancient pool of `primordial soup.'
"How do they believe this is possible? By millions and billions of accidents. For example, an early fish might accidentally grow a new kind of fin which helped him swim faster and escape his enemies. Then his fins might accidentally turn to legs he could use to walk on land, and so on.
"All this is based on a faith by the evolutionists that somehow, somewhere a gene changed to give this higher life form. It has to be faith, because there is yet no evidence that when genes have accidents (called mutations), that is for the better.
"The evidence is overwhelming that such accidents either make the gene worse or, at best, no better than the original.
"After all, how often do you see a car run faster and more smoothly after a head-on collision?
"Well, back to fruit flies. Because fruit flies reproduce many generations in a very short time, scientists picked them for the experiment hoping to compress thousands of years of `evolution' into a few years of lab work.
"After 80 years and millions of generations of fruit flies subjected to X rays and chemicals which cause mutations, all they have been able to produce are more of the same: fruit flies.
"And—more importantly—they have all been no better or stronger, and many have been weaker. All the changes eventually reached limits that, when approached, the strains of the fruit flies grew progressively weaker and died.
"And when the mutated strains were allowed to breed for several generations, they gradually changed back to the original form.
"One experiment produced fruit flies without eyes. Yet, after a few life cycles, flies with eyes began to appear. Some kind of genetic repair mechanism took over and blocked any possibility of evolution.
"God was very careful in Genesis to state that each of the animals were created `after his kind.' After 80 years and millions of generations, God was proven right: A fruit fly will always be a fruit fly."—"Evolutionists Still Looking for a `Good Accident,' " Battle Cry, July-August, 1990.
To the next topic in this series:
AN EVOLUTIONIST'S PARADISE: Paradise, for an evolutionist, is where he can go and see evolutionary changes occurring. Well, we've found the places where he should go.


text: OK so I have no need for another rifle seeing as I have quite a few on hand already. But I have never owned a Browning of any variety and I have always wanted one, my salesman today told me that they are having a sale on A-Bolt composite stalker rifles next month for $499! And just so happens that they will be carrying the Brownings in 7mm-08 which I have been wanting for some time as well. Just a few quick questions for the Browning crowd I know the A-bolt was built with the barrel "glued" to the receiver in the past making any attempt to re barrel the rifle nearly imposable, is this still the case? What are your feelings on the A-Bolt vs the X-Bolt as far as accuracy, overall feel, and reliability go? Are Browning synthetic stocks crappy molded injection turds like Remington/Marlin or are they a higher grade like the Sako/Tikka/Kimber? How is the trigger on the modern A-bolt?


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Azealia Banks Calls T.I. a 'F—ing Clown' for Continuing to Address Iggy Azalea Beef
Rapper Azealia Banks had some choice words for southern MC T.I. Monday afternoon (April 23). After word spread that the Trouble Man creator voiced his opinion, once again, during a radio interview regarding her beef with his artist Iggy Azalea, the 20-year-old "212" rhymer contacted Tip via Twitter. She proceeded to send him angry, expletive-laden tweets on the social networking platform because she wasn't keen on his comments.
T.I. sat down with WGCI's The Morning Riot to discuss his new LP in addition to the war of words Iggy and Azealia have been having as of late. According to Tip, Azealia should not be confronting him. "That is what I consider… bitch shit," he admits. "I'm a man. You ain't got no business addressing me. Get your man to address me. He and I can speak on it. Keep dealing with that woman [Iggy]. Me and you ain't got nowhere to go with that."
Tip shared that he knew the two lyrical ladies were "going through their thing" but he never saw tweets Azealia Banks directed at him when he first spoke of her verbal tiff with Iggy, who is signed to T.I.'s Grand Hustle imprint. After T.I.'s interview with The Morning Riot made its rounds on the Internet, Azealia took to Twitter once more to express her disapproval in him speaking about her actions.


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We now have both OpenOffice 2 and OpenOffice 3 on all the Linux machines.
For word processing, spreadsheets, slide shows, and the like you will be using the excellent OpenOffice programs. They will read your various Microsoft Office© documents and will write in the same format. (OpenOffice 3 will even handle docx)
Several instruction manuals are located in the library on the shelf over the computers or, if you are in the larger public access room, you can ask the staff member on duty for a quick lesson.
This software is very powerful, and if you like it and want to use it at home, you can download it yourself or go to the library and ask for a free copy.
See also [Write] | [Calc] | [Impress] | [drawing|Draw] | [Base] | [Math] | [software|other software]
Estamos encantados de informarle que ahora tenemos OpenOffice 2 y 3 en todas las computadoras operativas por Linux.
programs >
Subpages (1): templates