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Advances in large-scale analysis of human genomic variability provide unprecedented opportunities to study the genetic basis of susceptibility to infectious agents. We report here the use of an in vitro system for the identification of a locus on HSA8q24.3 associated with cellular susceptibility to HIV-1. This locus was mapped through quantitative linkage analysis using cell lines from multigeneration families, validated in vitro, and followed up by two independent association studies in HIV-positive individuals. Single nucleotide polymorphism rs2572886, which is associated with cellular susceptibility to HIV-1 in lymphoblastoid B cells and in primary T cells, was also associated with accelerated disease progression in one of two cohorts of HIV-1–infected patients. Biological analysis suggests a role of the rs2572886 region in the regulation of the LY6 family of glycosyl-phosphatidyl-inositol (GPI)–anchored proteins. Genetic analysis of in vitro cellular phenotypes provides an attractive approach for the discovery of susceptibility loci to infectious agents.
Author Summary
Individuals differ in their susceptibility to the HIV-1 virus, and the determinants of susceptibility are encoded in the human genome. Genetic variants influencing this trait have been identified by investigating candidate genes thought likely to be involved in HIV-1 pathogenesis or by whole-genome association studies, which type more than 500,000 genetic variants per individual (genome-wide association studies) to see which ones associate with susceptibility. We have addressed the issue of identification of new genetic variants influencing susceptibility to HIV-1 by a novel strategy based on the in vitro infection of cells. For this, immortalized B lymphocytes from 15 families (198 cell lines) were infected by a HIV-based vector. Differences in cellular susceptibility to infection—a genetic trait—could be mapped to a precise region on Chromosome 8, suggesting a role of the LY6 family of GPI-anchored proteins in HIV-1 infection. Genetic analysis of in vitro standardized cellular phenotypes provides a new approach to the discovery of the basis of genetic susceptibility to infectious agents.
Citation: Loeuillet C, Deutsch S, Ciuffi A, Robyr D, Taffé P, Muñoz M, et al. (2008) In Vitro Whole-Genome Analysis Identifies a Susceptibility Locus for HIV-1. PLoS Biol 6(2): e32. doi:10.1371/journal.pbio.0060032
Academic Editor: Hemant Tiwari, University of Alabama, United States of America
Received: July 12, 2007; Accepted: January 3, 2008; Published: February 19, 2008
Copyright: © 2008 Loeuillet 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: Funded by the Swiss National Science Foundation and the National Centers of Competence in Research (NCCR) "Frontiers in Genetics."
Competing interests: The authors have declared that no competing interests exist.
Abbreviations: CEPH, Centre d'Etude du Polymorphisme Humain; GFP, green fluorescent protein; GPI, glycosyl-phosphatidyl-inositol; LCL, lymphoblastoid B cell lines; LY6, lymphocyte antigen 6; SNP, single nucleotide polymorphism
Some individuals do not become infected to the HIV-1 virus despite repeated exposures, and among those that do, there is marked variation in the clinical course and progression to AIDS [1]. Although a number of host genetic determinants of susceptibility to HIV-1 have been identified through the analysis of candidate genes—most notably CCR5 Δ32 and HLA alleles—only a fraction of the observed phenotypic variation can be explained by variation at these loci [2,3]. Thus, there is a considerable interest in applying unbiased methods such as whole-genome analysis for the identification of novel susceptibility loci to human pathogens [1]. This hunt is, however, plagued by numerous confounding factors such as the lack of ascertainment of informative patient cohorts and difficulties to control for the variability of the infectious agent. Whole-genome mapping for viral susceptibility has been reported in mice for the murine adenovirus type 1 [4], and in mosquitoes for the dengue-2 virus [5]. Recently, the first genomewide association analysis for determinants of host control of HIV-1 in humans has been completed [6].
Whole-genome scans can also be performed through the analysis of family data using linkage analysis, an approach widely used to map monogenic disorders [7,8]. The need for family-based data has limited the use of this approach in the HIV-1 field because of the rarity, beyond instances of vertical transmission, of multi-case family infections. Studies of host genetic susceptibility to HIV-1 are also confounded by differences in virulence of the infecting viral strain [1].
To circumvent these limitations, we established an in vitro system to address the genetic control of cellular susceptibility to HIV-1 using cell lines from multi-generation families [9,10]. We used families from the Centre d'Etude du Polymorphisme Humain (CEPH) resource (up to four grandparents and an average of eight children per family), consisting of Epstein-Barr virus (EBV)–immortalized lymphoblastoid B cell lines (LCL). CEPH LCLs have been extensively genotyped, and the data are publicly available (http://snpdata.cshl.edu/population_studies/linkage_maps//). CEPH LCLs were previously used to identify genomic loci influencing sodium–lithium counter transport [11], natural variation in gene expression [12–15], transcriptional response to ionizing radiation [16], susceptibility to chemotherapy [17], and the relative impact of nucleotide and copy number variation on gene expression [18,19]. We hypothesized that CEPH LCLs could allow genome-wide investigation of interindividual variation of cellular susceptibility to infection with an isogenic virus, under standardized conditions and a controlled environment.
We designed the study to progress through consecutive steps including: (i) the identification by linkage and association of candidate markers associated with in vitro cellular susceptibility to an HIV-1-based vector, (ii) in vivo validation of candidate polymorphisms in humans infected with HIV, (iii) investigation of potential biological mechanisms (Figure 1).
LCL, lymphoblastoid B cell lines; HIV.GFP, vesicular stomatitis virus G protein-pseudotyped HIV-1-based vector; CEPH, Centre d'Etude du Polymorphisme Humain; SNP, single nucleotide polymorphism.
Since B cells are not a natural target of HIV-1, we established the conditions for efficient transduction of lymphoblasts with a VSV.G (vesicular stomatitis virus G protein)-pseudotyped HIV-1–based vector (HIV.GFP). We first assessed to what extent immortalized B cells reflect the behavior of CD4+ T cells by transducing purified CD4+ T cells and EBV-immortalized B cells, from the same 11 Caucasian healthy blood donors, with the same HIV.GFP. We used the green fluorescent protein (GFP) transgene expression as a reporter for permissiveness to lentiviral infection. We observed a significant correlation (Pearson r2 = 0.56, p = 0.007) between the level of transduction of CD4+ T cells and B lymphoblastoid cells for the same individuals (Figure S1). Thus, we hypothesized that transduction of B cells can capture a significant proportion of interindividual variation of post-entry events in the HIV-1 life cycle (reverse transcription, integration, transcription, and translation). Additional validation of the assay established the intra- and interday reproducibility of the transduction phenotype in CEPH LCLs, and ruled out an influence of potential confounders such as EBV copies per cell and the level of expression of the EBV-transforming protein LMP1 (unpublished data).
To determine whether variation in cellular susceptibility to the HIV.GFP virus has a genetic component we estimated heritability (h2, i.e., the proportion of variance attributable to additive genetic factors) in five CEPH pedigrees (76 individuals). In parallel, we also scored eight additional traits unrelated to HIV susceptibility (EBV copy number, EBV LMP1 oncogene CD11a, CD19, CD21, CD23, CD39, CD54). We observed a significant heritability of the HIV susceptibility trait (h2 = 0.54, p = 1.6 ×10−6), as well as for most of the other traits, with h2 values in the same range as those reported for gene expression variation traits [20] (Figure S2).
In view of these significant heritability results, we extended the analyses to 15 CEPH pedigrees (198 individuals) for the lentiviral cellular permissiveness trait (Figure S2), and we selected the expression of the endogenous cell surface marker CD39 (EBV receptor), and of the EBV-encoded LMP1 protein as additional study phenotypes.
To identify genomic loci that contribute to the variation in cellular permissiveness to HIV.GFP, we performed a quantitative genome-wide linkage analysis using a panel of 2,600 SNP markers with an effective resolution of 3.9 cM. Calculations were performed using the variance components analysis option from Merlin [21]. A region on HSA8q24 (marker rs1398296) showed the highest multipoint linkage score (logarithm of the odds [LOD] = 2.89, p = 1.3 × 10−04; Figure 2A). To determine the significance of this finding, we performed 500 simulations in which genotypes were randomized but the phenotype was kept constant, so as to preserve the heritability of the trait, the marker density, and the missing data patterns [22]. The distribution of maximum LOD scores of the simulations revealed that the observed HIV.GFP linkage peak is significant on a genome-wide basis at the 95% significance level (Figure 2A).
(A) Linkage analysis with 15 families and 2,600 markers identifies a quantitative trait locus (QTL) on HSA8q24.3. The highest multipoint LOD score of 2.89, p = 1.3 × 10−04 (marker rs1398296) was significant at genome-wide level as determined by permutation analysis (95% significance threshold of 2.83, dotted line).
(B) Association analysis in 56 unrelated individuals, using 521 SNPs from the HapMap project centered 3 Mb around the initial QTL, identifies marker rs2572886 (p = 1.8 × 10−5). The association remained significant after correction for multiple testing by Bonferroni (dashed line) or permutation analysis (dotted line).
(C) The candidate marker is associated with a significant 1.4-fold increase in susceptibility to the HIV.GFP.
(D) rs2572886A is associated with a significant 1.6-fold increase in susceptibility of CD4+ T cells from healthy blood donors to replicating HIV-1, differences between the alleles remain significant (p = 0.0331) after removing the outliers.
In order to independently confirm and fine-map the linkage analysis result, we assayed LCLs from 56 unrelated CEPH individuals that have been genotyped at a high density in the frame of the HapMap project [23]. The association analysis was performed using 521 tag SNPs in a 3-Mb region centered on the initial linkage assignment. A single SNP, rs2572886G>A, was strongly associated with HIV.GFP permissiveness (p = 1.8 × 10−5), and statistical significance was maintained after Bonferroni correction for multiple testing and permutation analysis (n = 10,000) (Figure 2B). Allele A of marker rs2572886 is associated with an average 1.4-fold increase in susceptibility to the HIV.GFP in LCLs from unrelated individuals, p = 0.001 (Figure 2C). Similar steps were taken for the secondary study phenotypes unrelated to lentiviral cellular susceptibility, which led to the precise identification (by linkage and followed by association) of a region involved in cis-regulation of CD39 expression (Figure S2). In contrast, no locus was identified affecting LMP1 expression, suggesting a more complex control of this trait by multiple genes.
Because observations were all made on B cells, we next assessed the potential role of rs2572886 as a susceptibility factor for HIV-1 infection in CD4+ T cells. We genotyped the SNP in a collection of purified CD4+ T cells obtained from 128 Caucasian healthy blood donors. CD4+ T cells were infected with a replicating HIV-1, and permissiveness was assessed by p24 antigen production [3]. A significant association was again obtained for SNP rs2572886 and cellular susceptibility to HIV-1 on this independent sample (p = 0.019) using a biological system that more closely resembles the in vivo situation. Consistent with the results of transduction of B cells with a HIV-1–based vector, in CD4+ T cells, the allele A of marker rs2572886 is associated with a 1.6-fold increase in susceptibility to infectious HIV-1 virus than CD4+ T cells of noncarriers, as assessed in a 7-d replication kinetics analysis (Figure 2D). The size effect associated with rs2572886 in vitro is comparable to that identified for other genetic variants influencing the HIV life cycle [3].
Since the previous results were obtained from in vitro assays, we set out to assess the potential association of rs2572886 with disease progression in HIV-1 infected individuals. The rs2572886 SNP has a minor allele frequency of 7% in Caucasians (Utah CEPH individuals), 19% in West Africans (Yoruba Hapmap sample), and 23% in Asians (Han Chinese and Japanese, HapMap sample). We genotyped 805 individuals recruited in the frame of the genetic project of the Swiss HIV Cohort Study (http://www.shcs.ch) who provided informed consent. These patients contributed consecutive CD4+ T cell data (n = 4,999 measurements) and viremia (n = 1,926 measurements) over an average follow-up period of 7 y in the absence of anti-retroviral drug treatment. The rs2572886A allele was associated with greater viral load, and faster progression of immunosuppression, as defined by the slope of CD4+ T cells depletion over time (Figure 3A and 3B). Individuals homozygous for the minor allele variant (n = 7) exhibited, as a group, a faster disease progression, but no conclusions can be made with such a small sample size. The same trends were also present in the incident cohort of 259 individuals identified within a 1-y interval of seroconversion (Figure 3C and 3D), although the limited numbers precluded significant association. These results are consistent with the in vitro data, since the A allele, which was associated with higher susceptibility of infection in the cellular systems, was associated with greater viral load and faster progression in vivo.
(A) Pattern of viral load in the full study population; each gray point represents a single viral load determination; the red (rs2572886A heterozygous), black line (individuals with the common allele), and orange lines (minor allele variant homozygous) represent the trend trajectories.
(B) Pattern of CD4+ T cell depletion in the full study population; each gray point represents a single CD4+ T cell determination.
(C) Pattern of viral load in infected individuals with known date of infection (incident sub-cohort).
(D) Pattern of CD4+ T cell depletion in the incident sub-cohort.
(E) Pattern of viral load in the validation incident cohort.
(F) Pattern of CD4+ T cell depletion in the validation incident cohort.
As an additional validation step, we genotyped a second independent cohort including 189 individuals with a precise date of seroconversion (Figure 3E and 3F), which was collected in the context of a whole-genome association analysis [6]. No association was detected in this cohort; however, the power to detect association in a sample of this size is estimated to be around 25%. These patients were recruited by eight different cohorts, while the original results were established using Swiss HIV cohort data. When pooling the discovery incident subcohort and the validation cohort, the association of rs2572886 on both the CD4 and viral load did not reach significance despite the increased number of samples to 448. However, the combined sample is far from homogeneous. Thus this association should be considered as suggestive at this point..
The association results should be discussed in the frame of the recently published genome-wide association analysis of host determinants of viral setpoint [6]. First, the marker identified in the current study, rs2572886, is neither present nor tagged by the Illumina HumanHap550 BeadChip used in the paper by Fellay et al. [6]. In addition, the design and premises of the genome-wide association and those of the genome scan reported herein are different: (i) the manuscript by Fellay et al. led to the identification of acquired/innate immunity loci (in major histocompatibility complex [MHC]) that cannot be captured in a cellular assay that investigates the viral life cycle; (ii) the study design in the paper by Fellay was powered to detect only very strong and sufficiently common genetic determinants; (iii) the standardized infection conditions and the study endpoint (expression of a reporter) that were used in the current study are very different from the conditions encountered in a population of HIV-infected individuals. It is through the nature of these profound study design differences that we aimed at generating complementary information to that provided by the genome-wide association analyses.
To provide a reference parameter that would allow comparison with genetic variants identified in other studies, we estimated the proportion of variation explained by rs2572886 and compared it to the contribution of CCR5 Δ32 in the same study population. Including CCR5 Δ32 into the model increased the proportion of variation explained by 1.9% for the CD4 cell count and 0.4% for the viral load. For rs2572886A, the estimates were 0.8% and 1%, respectively. For comparison, age at infection contributed to an increase in the proportion of variation explained of 1.4% for the CD4 cell count and 0.05% for the viral load, whereas the increase was 3% and 3.1%, respectively, for gender. These values are comparable to estimates indicated in the literature for various genetic variants influencing HIV pathogenesis [24], although direct comparisons are difficult due to different study designs. In contrast, the proportion of variation explained by rs2572886 or CCR5 Δ32 is considerable smaller than that for the genetic determinants reported in the study by Fellay et al., reflecting the fact that this genome-wide association study was powered to detect strong genetic effects. Thus, HCP5 and HLA-C variants explained 9.6% and 6.5% of the total variation in viral load, respectively, and an SNP near the RNF39 and ZNRD1 genes explained 5.8% of the total variation in disease progression [6].
The rs2572886 SNP is located in a nonconserved intergenic region on the telomeric end of Chromosome 8q (Figure S4). It is flanked on both sides by genes of the LY6/uPAR family (Figure S3). The LY6 genes are characterized by conserved cysteine-rich domains with specific disulfide bonding patterns but with little homology (20%–30% amino acid conservation among family members); members are either glycosyl-phosphatidyl-inositol (GPI)–anchored cell-surface receptors or secreted cytotoxins. Eight genes are located at 8q24.3: LY6K, SLURP1, LYPD2, LYNX1, LY6D, GML, LY6E, and LY6H. The functions of the encoded proteins are diverse but not well understood [25]. None has been associated with HIV-1 in the past, although the LY6H gene was reported to be up-regulated upon HIV infection [26]. A related protein of the LY6/uPAR family, the urokinase-type plasminogen activator receptor, coded by a gene in Chromosome 19, has been reported up-regulated in HIV-infected individuals, and proposed to participate in the innate immunity to HIV-1 through an interferon (IFN)-like mechanism [27,28].
The SNP rs2572886 is located in a recombination hot spot between two linkage disequilibrium blocks. We resequenced the surrounding region (∼ 13 kb) in 30 chromosomes to identify additional SNPs in linkage disequilibrium with rs2572886 that might point toward a biological function. Although two closely positioned SNPs—rs12546765 and rs12546801—were associated with rs2572886 in this limited resequencing dataset, they are not found in linkage disequilibrium in HapMap (pairwise r2 = 0.03). The region (∼1 kb) where rs2572886 is located is only present once in the human genome. We downloaded the homologous region of chimpanzee and Rhesus macaque and sequenced it in seven additional primates (bonobo, gorilla, orang-utan, nomascus gibbon, siamang gibbon, baboon, and African green monkey). rs2572886G>A was particularly variable among primates, with "G" representing the ancestral nucleotide in Old World monkeys, "A" the ancestral residue in hominoids, and "T" in gibbons.
To identify candidate genes that could be functionally related to the rs2572886 SNP, we performed quantitative 3C (chromatin conformation capture) [29], with the goal of detecting potential chromatin interactions between the SNP region and neighboring genes. We tested 11 regions by Taqman real-time PCR, spanning a distance of 190 kb surrounding the SNP. We focused primarily on the upstream areas (promoters) of genes in the locus. Results from cross-linked cells were compared to randomly ligated BAC DNA from the same region to correct for interassay differences and potential ligation biases. As expected, we observed a high level of enrichment with a region located 3.1 kb from the SNP (positive control) due to random chromatin interactions that have been reported to occur between regions separated by less than 5 kb [30]. The trend for enrichment rapidly decreased with increasing distance. Interestingly, we detected higher than background peaks of enrichment on the upstream areas of two genes—LY6D and LYPD2 (Figure 4)—suggesting that these are good candidates for functional interaction with the associated SNP. There are no apparent interactions between the SNP and the nearby GML promoter, despite its relative proximity (12 kb) in comparison to the LY6D (35 kb) and LYPD2 (70 kb) genes.
Curves show results of quantitative chromatin conformation capture (3C) for four independent CD4+ cell lines. The y-axis corresponds to interaction enrichment relative to background (furthest point) and corrected according randomly ligated BAC. The x-axis shows relative distance to rs2572886.
We prioritized the following proteins for additional biological assessment: LY6D and LYPD2 on the basis of chromatin conformation capture analysis, SLURP1 based on its unique status of secreted protein, and GML because of the proximity to the genetic marker. First, we overexpressed each of the four proteins from several vector backgrounds in 293T and HeLa cells to assess whether this would influence transduction by HIV.GFP. No significant changes in cellular infectivity were detected upon overexpression in these cell lines (Table S1).
In general, all eight genes of the LY6/uPAR family show detectable, but very low levels of expression as assessed by quantitative RT-PCR (unpublished); this precludes gene expression variation analysis to determine whether the genotype at rs2572886 correlates with expression levels of nearby genes. LY6D and LYPD2, which showed relatively higher expression levels, were silenced in HeLa cells by small interfering RNA (siRNA). Silencing with three different siRNA was successful for LY6D and suboptimal for LY6PD2. After transduction with HIV.GFP, we observed minor modifications in rates of cellular infection (Table S1). These findings are interesting, but given the cell type used and the harsh treatment of the cells, they are not conclusive enough to make a functional link for these genes. Overall, the biological basis for a role the LY6/uPAR family of proteins in HIV-1 cellular susceptibility remains elusive after this first line of biological screening. Additional analyses will be required to convincingly demonstrate a role for these proteins in the HIV life cycle.
In summary, by using a multi-step procedure involving a whole-genome linkage scan followed by association studies, we identified a locus on HSA8q24 that influences cellular susceptibility to HIV-1, and possibly progression of HIV-1 infection in vivo. Although the initial findings were based on transduction of transformed B lymphoblastoid cells with a HIV-1–based vector, subsequent experiments first on primary CD4+ T cells infected with replicating HIV, and second on a cohort of untreated HIV patients, supported the initial observations of association. In addition, although quantitative 3C data suggest a possible participation of genes of the LY6/uPAR family, further work is required to decipher the biological mechanism underlying this association.
Materials and Methods
Members of the LY6/uPAR family—LY6D (GenBank NM_003695), LYPD2 (NM_205545), GML (NM_002066, SC303114; OriGene), Lynx1c (NM_177457), Slurp1 (NM_020427), and Slurp2 (NM_177458)—were amplified by RT-PCR from RNA extracted from human cell lines (HeLa or 293T), and cloned into pCI-neo. LY6D and LYPD2 were tagged C-terminally with an HA tag.
Cell culture.
293T cells were cultivated in Dulbecco's modified Eagle Medium (DMEM; Invitrogen) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 50 μg/ml gentamycin. 293T cells (3 × 106 cells) were cotransfected with 20 μg total of DNA (empty pCI vector + increasing amounts of pCI vector containing the gene of interest) using the calcium phosphate technique. Twelve hours post-transfection, cells were washed and 300,000 cells were seeded in six-well plates and incubated further for 36 h to allow the expression of the gene of interest before HIV-based vector infection.
CEPH cell lines.
EBV-transformed B cell lines were obtained from the CEPH collection through the Coriell Institute for Medical Research (http://locus.umdnj.edu/nigms/ceph/ceph.html). Cells were cultivated in RPMI 1640/Glutamax-I medium (Invitrogen) supplemented with 15% fetal calf serum (FCS, Inotech). They were maintained by replacing half medium twice a week. Pedigrees studied were numbers 102, 884, 1328, 1331, 1332, 1333, 1334, 1340, 1341, 1345, 1346, 1347, 1362, 1408, and 13292.
CD4+ T cell isolation and B cell immortalization.
Cells from 11 white healthy blood donors were used to isolate CD4+ T cells by using anti-CD4 magnetic beads (Miltenyi Biotech). Cells were cultured in RPMI1640/Glutamax-I medium supplemented with 20% FCS, 20 U/ml human interleukin-2 (IL-2, Roche) and 50 μg/ml gentamicin (Invitrogen) following stimulation with phytohemagglutinin (PHA) at 2 mg/ml for 2 d [31]. The CD4-negative cell fraction was exposed to EBV containing supernatant from a B95–8 cell line according to current protocols [32].
HIV-based vector production.
To produce HIV-based vector particles (HIV.GFP), 293T cells (3 × 106 cells) were cotransfected with four plasmids using the calcium phosphate method. Plasmids encoded the VSV-G pantropic envelope (pMD.G), the Gag and Pol proteins (pCMVΔR8.92), Rev (pRSV-Rev), and the fourth plasmid encoded the HIV vector segment carrying GFP as the reporter transgene under the control of the CMV promoter (pWPTS-GFP) (kind gifts from D.Trono, EPFL, Lausanne, Switzerland; see http://rd.plos.org/pbio.0060032.1 for vector details). Forty-eight hours after transfection, the supernatant was collected, centrifuged to pellet cellular debris and filtered through 0.45-μm filters. Viral particles were concentrated by centrifugation through a 100-kDa cut-off membrane (Centricon Plus-70; Millipore AG).
HIV-based vector transduction (single-round infectivity assay).
Transduction of CEPH cells and of the B cells of healthy blood donors (0.5 × 104 cells in 96 wells) was performed by spinoculation with HIV-based particles for 3 h at 1500g and 22 °C. After 72 h, cells were harvested and expression of GFP protein was monitored by fluorescence activated cell sorting (FACS) analysis with mock transduced cells as control. This was performed twice in triplicate at 1-wk intervals. Similarly, transfected 293T cells (in six-well plates) were infected with HIV-based particles (25 ng p24 equivalent) in the presence of 10 μg/ml DEAE-dextran in 1 ml D-10 for 5 h. Culture medium was replaced and cells were incubated for 24 h before FACS analysis of GFP expression. Typically, 20%–30% of GFP+ cells were obtained for controls.
HIV infection.
Healthy blood donors' CD4+ T cells (106 cells) were infected with NL4-3BaL virus (1,000 pg of p24 antigen) in a 1-ml final volume for 2 h at 37 °C in 5% CO2. Cells were washed and cultured for 7 d. Virus-containing supernatant was harvested, and p24 antigen production was monitored by an enzyme-linked immunosorbent assay (ELISA) (Abbott).
For cell surface molecule staining, 105 CEPH cells were washed, resuspended in PBS/0.5% BSA (Sigma) and incubated with primary monoclonal antibodies (mAbs) or isotypes for 15 min at room temperature (RT). Primary mAbs were : anti-CD11a (Dako, MHM24), -CD19 (Dako, HD37), -CD21 (Dako, 1F8), -CD23 (Dako, MHM6), -CD39 (Serotec, A1), -CD54 (Dako, 6.5B5), and the negative control was mouse IgG1FITC (Dako, X0927). All mAbs were used at 1/50, except CD19 used at 1/20. For intracellular staining, CEPH cells were washed and resuspended in cytofix/cytoperm solution (Becton Dickinson) for 20 min at 4 °C. After two washes with permwash, cells were resuspended in permwash with primary anti-LMP1 (1/50, S12, a gift from S. Rothenberger) or negative control antibody mouse IgG2a (1/50, Dako), for 15 min, at RT. After washes, cells were incubated in permwash with secondary anti-mouse PE (1/30, Dako). After wash, cells were fixed (CellFix, Becton Dickinson) and analyzed using a FACSCalibur system for 10,000 events. Positive events were defined as a fluorescence level superior to that of isotypic control. Determination of EBV copy number was carried out by real-time PCR by using specific probes as described [33].
Heritability, linkage, simulations, and associations studies.
Heritability calculations (h2) were performed using the "polygenicscreen" command from the SOLAR software [34]. SNP genotyping data, consisting of 2,688 autosomal SNPs were downloaded from the SNP Consortium database (http://snpdata.cshl.edu/population_studies/linkage_maps/) [35]. Multipoint linkage with the SNP map was performed using Merlin [36] with the –VC option, after Mendelian inconsistencies (PEDCHECK) [37] and unlikely genotypes (PEDWIPE) [38] were removed. To calculate the empirical significance of the linkage results, we performed 500 simulations for each quantitative trait using the –simulate command from Merlin with different seed numbers. We extracted the highest result from each simulation to build significance distributions. All simulations were performed using a cluster of 32 HP/Intel Itanium 2 based servers at the Vital-IT Center (http://www.vital-it.ch/). Association analysis of quantitative phenotypes (% of GFP-positive cells and / mean fluorescence intensity (MFI) of CD39), and corrections for multiple testing were performed using the PLINK software (http://pngu.mgh.harvard.edu/~purcell/plink/anal.shtml). Genotypes were downloaded from the HapMap project URL (http://www.hapmap.org/cgi-perl/gbrowse/gbrowse/hapmap/), HapMap public release number 19.
Cohort analysis.
Data from both incident (patients identified during primary infection or who have had a negative and positive test for HIV infection within a narrow time interval, 1 y in this study, in which case the date of infection is estimated as the mid-point), as well as data from prevalent cases (i.e., individuals already HIV-seropositive by the time they entered the study, unknown date of infection) were analysed longitudinally by modeling the CD4 T cell count and HIV-1 RNA marker's trajectories over time for the different genotype groups. The analysis was conducted using population-averaged marginal modeling [39], because the focus of the study was to investigate the effect of specific genetic factors on disease progression at the population level. In a marginal model, the mean regression function is modeled independently from the variance–covariances matrix. We used fractional polynomials to assess the best-fitting functional form. The viral load (log scale) and the CD4 (square root scale) trajectories post seroconversion were linear and appeared stationary. Therefore, linear functions of time, along with interactions with polymorphisms and covariables (age at infection and gender) were considered. The impact of genotype on slope and intercept was assessed using Wald test, and the proportion of explained variation was assessed [40]. A multivariate distribution was fitted to the data by score-like methods (generalized estimating equations) [41]. The correlation structure was assumed to be well represented by an autoregressive process of order 1. To limit the impact of frailty selection, only the data for the first eight years since seroconversion were considered. The analysis was repeated considering, in turn, only the incident, prevalent, and both cohorts. Subanalyses were also performed considering the Caucasian group only. For the prevalent cases, an estimate of the unknown date of infection was obtained using the markers data and defining for each patient an infection window based on his or her last negative and first positive available HIV tests. The date of infection was then imputed using a methodology that extends published methods [42,43] to accommodate multiple marker measurements per individual (P. Taffe and M. May, unpulished data, and [44]. A second incident cohort, recruiting individuals from various European countries, was used to validate results obtained from the analysis of the incident cohort recruited within the Swiss HIV Cohort study. Statistical analyses were conducted using SAS version 9.1 for Windows, as well as STATA 9.2 .
Re-sequencing in human and nonhuman primates.
The region (∼1 kb) around the candidate marker was resequenced by using forward primer SG2000 (5′-AGTTCATACCCCTTTGCCAGGTTG) and reverse primer SG2001 (5′-GAAGCCTTACCTGCTTCCTGCC), and forward primer SG1829 (5′-TTCCCTGAGCTTGCAGGACTC) and reverse primer SG1853 (5′-CTCTACACACCTACCTTGCTGGGA) to generate overlapping PCR products.
Sequences have been submitted to GeneBank for bonobo (EU340888, Pan paniscus), gorilla (EU340889, Gorilla gorilla), bornean orang-utan (EU340890 , Pongo pygmaeus), nomascus (EU340891, Hylobates leucogenys), siamang (EU340892, Hylobates syndactylus), baboon (EU340893 Papio hamadryas), and African green monkey (EU340894, Cercopithecus [chlorocebus] aethiops).
Quantitative chromatin conformation capture.
Approximately 107 stimulated expanded primary CD4+ T cells [31] were crosslinked in their media for 10 min at RT with 1% formaldehyde (v/v). Crosslinking was quenched with 125 mM glycine prior to two successive washes with 1xPBS. Pelleted cells were resuspended into 5-ml ice-cold lysis buffer (10mM Tris HCl, pH 8.0, 10 mM NaCl, 0.2% (v/v) NP-40) complemented with protease inhibitors (Complete, Roche) and 0.5 mM PMSF. Lysis of the cells was allowed to proceed for 10 min on ice with mild shaking. Nuclei were recovered by centrifugation (600g for 5 min at 4 °C) and resuspended into 500-μl 1.2xDpnII restriction buffer (NEB). Nuclei were lysed with 0.3% (v/v) SDS for 60 min at 37 °C. SDS was sequestered with 2% (v/v) Triton X-100 for another 60 min at 37 °C. Chromatin was subsequently restricted overnight at 37 °C with 500 U DpnII (NEB) in a final reaction volume of 600 μl. After heat inactivation of the restriction enzyme (10 min at 65 °C), chromatin was dialyzed (Slide-A-Lyzer, Pierce) for 1 h against 1.5 l of water at RT and transferred into 7 ml ligation reaction mix (50 mM Tris HCl, pH 8.0, 10 mM MgCl2, 0.5mg/ml BSA, 10mM β-mercapto-ethanol, 0.5 mM ATP and 400 U T4 DNA ligase (NEB). The ligation reaction was performed for 4 h at 16 °C followed by another 30 min at RT. Crosslinking was heat-reversed and proteins were degraded (300 μg proteinase K) overnight at 65 °C in a hybridization oven. DNA was purified by phenol/chloroform/isoamyl alcohol [25:24:1(v/v)] extraction, precipitated with isopropanol, and washed with ethanol 70%. DNA was subsequently resuspended with 200 μl 1xTE pH 8.0 and treated with 50 μg RNaseA for 30 min at 37 °C. Finally, DNA was extracted with 1 volume phenol/chloroform/isoamyl alcohol [25:24:1(v/v)], ethanol precipitated, and resuspended into 100 μl 1xTE pH 8.0. Cross-linking was independently performed on four CD4+ lines derived from different individuals, two of whom (2 and 4) were heterozygous for rs2572886.
For quantitative Taqman PCR, we designed 11 assays comprising the PCR primers and a dual-labeled probe sitting at the predicted DpnII junction between the target and bait regions (primer and probe sequences are available upon request). Reactions were set up using a Biomek 2000 robot (Beckman), in a 10-μl volume in 384-well plates. Three replicates per assay per sample were performed. PCRs were run in an ABI 7900 Sequence Detection System (Applied Biosystems) with the following conditions: 50 °C for 2 min, 95 °C for 10min, and 50 cycles of 95 °C 15 s/60 °C for 1 min. Each reaction contained 300 nM of each primer and 250 nM of probe.
For the 3C samples, approximately 200 ng of DNA was used per well, and for the BAC (digested – randomly ligated) samples, 10 ng of DNA was used. Normalization for each assay was performed using the values obtained from BAC experiment (all assays are expected to give the same result, given that the naked BAC was fully digested and re-ligated, and all ligation combinations are expected to be present equimolarly). Enrichment was calculated with respect to the most centromeric probes, which showed very low levels of interaction.
Note in Press: Recently, Brass et al [45] identified through a siRNA screen over 250 HIV-dependency factors. Among these there were three members of the LY6/uPAR family (GML, LY6D, and LYPD4). This new evidence provides independent support for a biological role of the LY6/uPAR family in HIV-1 pathogenesis.
Supporting Information
Figure S1. Validation of the CEPH Model to Study HIV-1 Susceptibility
CD4+ T cells and B cells from same healthy blood donors were transduced with the HIV.GFP. Linear regression of one representative experiment for the percentage of GFP-positive cells is shown. Each point represents the mean of triplicate values.
Figure S2. Expression and Heritability of HIV-1 Susceptibility Trait in CEPH Families
(A) Heritability estimate (h2) for eight study traits (percent of positive cells and mean fluorescence intensity, MFI), and for the EBV copy number (** p < 0.001, *** p < 0.0001).
(B) Representation of interindividual (black dots), and interfamily (red bar = median value for each family) differences in percentage of GFP-positive cells after transduction with a HIV.GFP of 198 lymphoblastoid B cell lines from 15 CEPH pedigrees.
Figure S3. Genome Scan and Fine Mapping of CD39 Expression (MFI Trait)
(A) Results of linkage analysis with 15 families. A single locus on HSA10q23 (rs766083) is significant after correction by permutation analysis (n = 500).
(B) Results of association analysis in 56 independent individuals using SNPs from the HapMap project. All SNPs in a 700 kb region surrounding the CD39 gene were used. rs numbers next to peaks indicate significant SNPs after correction for multiple testing.
Figure S4. Chromosomal Localization of rs2572886
The SNP is localized in an intergenic region containing genes belonging to the LY6/uPAR family. The figure shows images generated through the University of California at Santa Cruz genome browser and Haploview software.
Table S1. Infectivity of HIV.GFP of Cells Transfected with or Silenced for the Gene of Interest from the LY6/uPAR Family
Values reflect relative infectivity to that of the mock control. None of the differences are statistically significant.
We thank D. Hohl and B. Favre for discussion and for SLURP1 reagents; R. Lubomirov, E. Atanga, M. Ortiz, and R. Martinez for assistance with re-sequencing, primate sequencing, genotyping and analysis; J. Fellay for assistance with setpoint analysis; and the Swiss HIV Cohort Study and EuroCHAVI physicians for permission to genotype the study SNP.
Author Contributions
CL, SD, AC, DR, JSB, SEA, and AT conceived and designed the experiments. CL, SD, AC, DR, and MM performed the experiments. CL, SD, AC, DR, PT, SEA, and AT analyzed the data. CL, SD, AC, JSB, SEA, and AT wrote the paper.
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text:
Utahns anxiously awaiting details on federal budget cuts
If federal funding is indeed reduced to those programs, he said he's not expecting the state to make up the loss, especially after several tough budget years.
"There's no alternative but to decrease services unless someone wants to come up with more revenue from state sources," Shumway said.
Gov. Gary Herbert's budget director, Ron Bigelow, said the state has already been preparing for reductions in federal funds, but he, too, is waiting to see what effect the debt-reduction deal will have on Utah spending.
State agencies are already required to halt spending for programs paid for with federal funds if that funding ends. And a new state law requires agencies to have contingency plans in place by October for reductions in federal funding.
Bigelow said the state expects a surplus from the budget year that ended June 30 on the "high side" of an earlier estimate of between $10 million and $110 million. Plus the state has a $200 million Rainy Day fund.
Still, he said, it would be up to lawmakers to decide whether to restore any federal cuts to the state budget, either in a special session or during the 2012 Legislature.
"A lot of people would like the state to step in and solve every problem. But, of course, we don't because we also have an obligation to the taxpayer," he said. "We can't keep going to people who are just barely getting by and say, 'Pay more taxes.'"
Rep. Ken Ivory, R-West Jordan, the sponsor of the legislation requiring contingency plans for the loss of federal revenue, said lawmakers will be "boring and prudent" in dealing with any federal cuts.
"We're still going to take care of our poor people in Utah. We're still going to take care of our roads. We're still going to educate our children," Ivory said. "That's just Utah values. That's just who we are. That's just what we do."
Contributing: Marjorie Cortez
E-mail: firstname.lastname@example.com
Twitter: dnewspolitics
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subset: commoncrawl


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Video Industry News Video Company News Technology Industry News Technology Company News Novelty Industry News Novelty Company News Gay Industry News Gay Company News Legal Industry News All Company News
Released: Jan 01st, 2005
Running Time: 96
Director: Dcash
Company: Dsynternet Media
Cast: Kiwi, Bianca (I), Stallion (I), Stephanie (I), Slick, Jah T, Jezebel (I), Adina Jewel
Critical Rating: AAA 1/2
Genre: Pro-Am
Black amateurs.
Few things in porn are as sweet as that innocent girl who shows up thinking she's just going to be taking a couple of test photos and ends up fucking on camera. When Jezebel shows up at Dcash's office, she is pretty firm about not being ready to shoot on camera. But performer Jah T. steps into the office and starts salivating over the prospect of fucking her. He just isn't going to let her leave before he fucks her. Jezebel gives in and ends up draining Jah T.'s nuts all over face and down her throat. By the end of the scene, the tables have turned, and the girl doesn't want to let go of Jah T.'s cock.
A couple of other big booty girls - with asses liberally oiled up with baby oil - help keep the eyebrows raised, particularly Bianca, a dark chocolate girl with a giant ass, and Kiwi, a slut with one bad attitude.


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A Rather Refined Palette
My name is Brian Jun, otherwise known as EU03 elsewhere on the Internet. I'm currently intern-turned-assistant art monkey for the rest of the art department at Reverge Labs.
Being one who can fumble through digital imaging, I'm assigned some of …Continue Reading »
Skullgirls, GGPO… and You
Yo – let's talk about GGPO.GGPO, short for "Good Game, Peace Out" is a networking library written by Tony Cannon (of shoryuken.com and Evo fame), and it's what Skullgirls uses for online play. I personally believe it is the… Continue Reading »
Reflecting on Evo
Mike Zaimont
EVO is all a blur… was I even at my panel? Because I'm pretty sure I was still asleep.
Even though it keeps happening, it never feels less weird to be asked to sign things.
I watched …Continue Reading »


text:
The Evolution of RightsWritten by Kurt Williamsen
I find it interesting that many people believe they have a right to demand "rights" — such as jobs, welfare, and healthcare — in the United States even if they are present here illegally. Disregarding for now the rightness or wrongness of their claim of rights, I begin to wonder, "Where will their rights end?" If the only real claim that people have to being entitled to stuff, stuff that must be paid for by other people who are forced by government to "donate" cash to the cause, is that they reside in the country (again, illegally) and that it's unfair for them to go without something that others have, then doesn't it seem that there ought to be many more rights?
Last week the President of Mexico set off on a human rights lecture tour of Washington, D.C. and chief among his claque of foot tappers was the President of the United States.
Florida this year sent to the Congress of the United States its application for a convention for the purpose of proposing a balanced-budget amendment to the U.S. Constitution. Florida is, by some counts at least, the 33rd state to apply. But the legislatures of New Hampshire and South Dakota have recently voted to rescind their applications. So are we now three states away from the 34 needed to require Congress to call a convention? Or one? Or 15 or so (arrived at by subtracting all the states that have rescinded their calls over the past 22 years)?
Linda McMahon, a Republican candidate for U.S. Senate in Connecticut, said Wednesday night that she did some of the research for what became a front-page story in the New York Times about how Democratic candidate Richard Blumenthal falsely claimed to have served in Vietnam.
Can't Keep Ignoring the SignsWritten by Raven Clabough
Most Americans recognize that the United States is currently facing major struggles, and that America is undergoing a fundamental transformation, but of those Americans, many still do not subscribe to the theory that there is an underground movement toward global government. The problem with that denial is the movement has come out from underground and has become so blatant that it can no longer be ignored or refuted.


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Cambro Portable Bars
Serve Delicious Drinks Wherever Your Business Takes You with a Portable Bar from Cambro
The Cambro portable bar lets you take the party anywhere! The easy mobility of these bars allows you to provide beverage and bar service in a variety of locations. Perfect for weddings, banquets, parties, and other catered events, Cambro's portable bars let you bring a superior happy hour or reception to your guests. A portable bar should have all the functions and storage your bartender needs to keep your guests happy and hydrated, and the Cambro portable bar does not disappoint! Read more
Many of these portable bars are equipped with features like speed rails and towel rings so that your bartender can quickly mix and serve your signature drink while keeping messes and spills at bay. Many of the Cambro bars are equipped with an ice sink to limit necessary refills so your bartender can keep the drinks flowing!


text: On Sunday, Texans shopping with online retailer Amazon began paying sales tax on their purchases, thanks to a deal between the state and the company. The deal came after the state billed Amazon millions for sales taxes on past sales to Texas buyers. Though consumers may not like it, it does end an unfair advantage Amazon had over Texas retailers.


text:
Awesome F-22 Raptor Screen SaverFree
Quick Specs
Total Downloads:12,392
Date Added:Jan. 23, 2002
File Size:1.13MB
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From Cottonwood Software: A photo slideshow screen saver with photos of U.S. Air Force aircraft. Includes fighter jets, bombers, and experimental aircraft from the A-10 to the X-29. All photos are 800x600 high-quality imageA photo slideshow screen saver with photos of the F-22 Raptor aircraft. See the Raptor firing missiles, refueling, taking off, climbing, banking, flying formation and more. All photos are 800x600 high-quality images. (1024x768 also available, see our web site.) Other screen saver features include transition effects, black and white mode, password use, and extract photos as wallpaper, including built-in wallpaper cycling every hour or every day. This Lite version with 5 photos is free, the Gold Edition has 30 photos and may be purchased at Awesome-Screens.com.
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text:
Definition of rouleau in English:
Late 17th century: French, from obsolete French roule 'a roll'.
For editors and proofreaders
Syllabification: rou·leau
Definition of rouleau in:
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= trending


text:
4-4-14 Track and Field: Tiger Track Classic Day 1
4-4-14 Track and Field: Tiger Track Classic Day 1
2014 Outdoor All-American (javelin)
2014 All-SEC Second Team
2014 SEC All-Freshman Team (javelin)
2015 Outdoor Season:
Competed in seven meets in the javelin ... Recorded a throw of 219-9/66.99m at the NCAA Prelims ... Threw 222-8/67.87m at the SEC Championships to finish eighth ... Placed second at the Border Clash with a throw of 203-8/62.08m ... Placed third at the Tigers Track Classic at 203-4/61.98m.
2014 Outdoor Season:
Snapped the school's 18-year record in the javelin at the SEC Championships with a distance of 239-7/73.04m ... Finished 13th at the National Championships in the javelin with a throw of 215-8/65.75m ... Named a Second Team All-American ... Qualified for NCAA Championships after finishing 10th at the NCAA East Preliminaries with a throw of 222-6/67.82m ... Broke the school's freshman record in his first meet with a throw of 223-11/68.26m ... Named to the Men's All-SEC Second Team as well as the All-Freshman Team.
Prior to Auburn:
Born July 29, 1995 ... Attended nearby Auburn High School in Auburn, Alabama ... Won javelin throw at 2013 New Balance Nationals with a long throw of 212-7 ... Mark ranks third nationally and was the best throw in Alabama ... Won 2013 Alabama State Championship with a throw of 197-6 ... Threw 175-11 in 2012 to place second at the state meet ... Has only thrown javelin for two years ... Threw 211-7.75 at the Chicagoland Throwers Series in June 2013 to place sixth ... Top throw ranks second all-time in Alabama history ... Named an All-American in 2013 for his performance at New Balance Nationals.
|Carter's Personal Records
|2014 SEC Outdoor Championships
* Denotes school record


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If we run the wildcat it means that Ben either leaves the field or lines up at WR. Is that really what we want to do on O?
If we run the wildcat it means that Ben either leaves the field or lines up at WR. Is that really what we want to do on O?
the wilcat works, but like just mentioned, why would we run it when we have a Hall of Fame qb..........
If we ran it, I would rather he left the field altogether. Not worth the risk of injury. And have Dixon run it. At least there is the threat of a pass in that situation.
That Wildcat looks a lot like plain and simple option football.
When will we see the wishbone?
Personally, I think with our fast defense, we handle the wildcat just fine.
6- Time Super Bowl Champions......
2012 MNF Executive Champion
okay, time for my dumb question of the day.....
why is it called the wildcat???? was a college team that first adapted this named the wildcats....
Ben was a WR in high school until his senior year because the coach wanted his son to be QB. If Dixon were Tomlin's son, then I would be worried.Originally Posted by steeler_fan_in_t.o.
1 CB Vernon Hargreaves Florida 5-10 204
2 DT Austin Johnson Penn State 6-4 314
3 SS Jeremy Cash Duke 6-0 212
4 LB Jaylon Smith Notre Dame 6-2 223
6 WR Moritz Boehringer Germany 6-4 227
7 OT Brandon Shell South Carolina 6-5 324
7 RB Tre Madden USC 6-0 223
DE David Onyemata, LB Ian Seau, FS Jordan Lomax, CB DeAndre Elliott,
QB Jeff Driskel, TE David Morgan, OG Spencer Drango, OC Mike Matthews
2015 MNF Executive Champion!
Originally Posted by Shoe
We do have that with Mendenhall he just needs to get the carries and as for a previous comment we have a guy that can run and throw if Arians chooses to run the wildcat in Dixon. The steelers won't run the wildcat tho.
That is very true.Originally Posted by JTP53609
2) I too, think he handle Miami's Wildcat better than most.
3) As much as anything else, I think I started the thread to ignite a little pro-Tebow mindset. I don't necessarily buy into his "Angel" image (i.e. I can't imagine how someone that age could eschew unlimited Florida-fed ).
But he is definitely an NFL caliber athlete. Add to it his (QB) experience. AND maybe most importantly, his Joey Porter/Ray Lewis like motor & intensity--that's an underrated quality IMO.
Guys and gals,
If you play fantasy football, go to the FF forum here and read my thread. I'd appreciate some feedback on it.


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I also prefer pampers diapers and have purchased a small bunch I'd them already also and plan on stocking up on more before April.
Question is....
1. There was no such thing as a boppy it a bumbo seat when I had babies. Do you recomend them as much as a few friends have?
2. What's your favorite brand of what? Just curious :)


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Forgotten Doll Ball Jointed AA by cdlitestudioWatch
©2011-2016 cdlitestudio
Awesome, just awesome
I agree, it really does look like GaGa O.O
She reminds me off mother monster, lady gaga...love it!
Your work always amaze me, I didn't know you have deviant web, I follow you on facebook, your're a person with a big heart too.
love your werk
She looks stunning. O_O
How did I miss this?
Very pretty Connie!!
Really great!
can't wait to get back to sculpting.. great sculpt!!
looks kinda like Lady gaga..but wow this is amazing!!! its very beautiful..keep it up!!!
I don't know what it is but I think you've outdone yourself with this one, especially with that wig. Your photos and presentations are getting better too.
WOW! Awesomeness!
your dolls are amazing!!! I can't find any other words to describe them. a real piece of art.
I love her!!!!!!!!!
Great job. Love the atmosphere she has.
So amazing. She looks so real. The look on her face is stunning.
sweet job
Wow, wow, wow, very beautiful.
Her face is so beautiful.
Her face is so beautiful.
I really love her make-up~ Her pose and everything in this shot is brilliant too!


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SEOUL, July 11 (Yonhap) -- Sales of foreign luxury brands dropped in South Korea in their fiscal 2012, data showed Thursday, due mainly to the protracted slump in the local economy, which dented consumers' sentiment.
According to the data compiled by the Financial Supervisory Service, Burberry Korea saw their sales drop 5.3 percent on-year to reach 228.1 billion won (US$201.5 million) in fiscal 2012. The firm closed its books on March 31.
Net profit of the British brand plunged 35.1 percent to reach 16.8 billion won over the cited period, the data showed.
Other designer goods also lost ground in fiscal 2012, with Gucci Group Korea Ltd. posting 282.6 billion won in sales, down 4.5 percent from a year earlier. The company closed its book on December 31.
Market watchers said the stagnant performances posted by luxury brands operating in South Korea came as the protracted economic slowdown gave local consumers limited leeway in making extravagant purchases.
Ferragamo Korea Ltd., whose parent company is Salvatore Ferragamo Italia SpA, also saw its net income drop 12.8 percent over the cited period, despite that its sales edged up 1.2 percent to 98.4 billion won.
Prada Korea Ltd., a unit of Prada SpA, on the other hand, saw its net income advance 10.5 percent on-year to reach 58.8 billion won in fiscal 2012. The number, however, slowed from a 64.2 percent growth tallied in fiscal 2011.
<All rights reserved by Yonhap News Agency>


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The Republican National Convention will be held at Quicken Loans arena in in Cleveland, Ohio from July 18 - 21 and it marks the first time the Ohio city has hosted the event since 1936.
If you have an event you would like to submit, send info about the event and a phone number to email@example.com 1 Eat like a child The Depot Theater's youth summer production ends Saturday with How to Eat Like a Child by...
The Kansas Humanities Council has named Dodge City resident Jane Holwerda to their board of directors. Holwerda would be the second Dodge City resident to be part of the board. According to KHC associate director, Tracy Quillin,...
For more than a decade the last Saturday night of Dodge City Days a free classic rock show has been held at the band shell in Wright Park. This year fans will notice a difference. "Jury is not playing this concert," said Donnie...
We hope the
Once Upon a Time is adding a new face for Season 6, but he won't be from the
The first game between the Dodge City A's and Derby Twins was moved to Saturday due to inclement weather. The A's and Twins will make up Friday's game by playing a doubleheader. Game one is scheduled for 6 p.m. at Cavalier Field.
Daryn Pittman dominated the Knight Before the Kings Royal at Eldora Speedway on Friday night, as he cruised to his sixth World of Outlaws Craftsman Sprint Car Series victory of the season.
Spoiler warning for Season 6 of The Walking Dead, The Walking Dead comics, and maybe Season 7, we'll see. In the Season 6 finale of
Ford County Directory
Business Name
Location, ST | website.com


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Thanks for this plugin, it's fantastic!
After my long list of research almost 12 hour I ended up with this plugin i must confess a good effort made by developer and i hope to see the update in later days.
Keep up mate and thanks for the plugin.
Manoj Karmocha
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superb plugin
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Legacy Benefits, LLC is an innovative specialty-finance firm focused on the life settlement market. The company identifies, evaluates, acquires and services inMore
$1 mil. - $5 mil.in Revenue
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The Lancaster Dixie Baseball AAA all-stars finished third in the state tournament after being eliminated by a red-hot Blythewood team on Wednesday night at the Timken Complex in Union.
Blythewood, the lone unbeaten in the state field, used blazing bats to score 11 runs in the first inning and added eight more in the second frame in taking a 19-0 win over the Lancaster stars.
Lancaster opened the double-elimination tournament with a 9-4 loss to Sumter, but reeled off three straight wins before falling to Blythewood.
Lancaster Super Speedway will be playing host to the USCS Winged Outlaw Sprint Cars Summer Shootout Round 4 on Saturday night.
Some of the country's top 360 Sprint Car drivers are set to invade the LSS big half-mile where the cars will bring their 140-plus mile per hour Outlaw cars.
The field will be led by the ever-popular Eric Riggins Jr., Brandon McLain, Terry Gray, Morgan Turpen, Nick Snyder, Jake McLain, Brandon Taylor, Danny Sams III, Lance Moss, Derek Hagar and Johnny Bridges.
The Lancaster Dixie Baseball Jr. Dixie Boys (13U) All Stars got off to a red-hot start in state play before back-to-back losses in the double-elimination tournament knocked them from contention.
On Saturday, the local stars topped perennial power Sumter, 6-4.
The Lancaster stars then fell to Bamberg, 14-11, on Sunday in the second round and were eliminated Monday in a 5-3 loss to Hilton Head.
Lancaster manager Bryant Neal said his team had its chances Monday, but couldn't get runs across the plate.
David Oliver continued his stellar play in the Lancaster chapter of the S.C. Junior Golf Association tournament which was played July 11 at the Chester Golf Club.
Oliver shot a five-over 41 over nine holes to take a five-shot win.
Oliver's stellar effort included two pars to go with a birdie. Oliver held a 45-point lead in the boys 10-12 age group after the Chester tourney.
Braden Small posted a 46 to place second, which included a par.
Toby Willis and Luke Benfield each carded a 51 to place in a tie for third.
AIKEN – The Lancaster Dixie Boys U14 all-stars team saw its state championship hopes dashed with an 11-6 loss to an undefeated Hampton team on Tuesday night at Citizens Park.
Hampton, of the Edisto area, built an 8-1 lead and held on for the five-run victory.
"I'm proud of these guys because after they fell behind 8-1 in the game tonight [Tuesday]. They showed a lot of grit and battled all the way to the final out," said Lancaster Dixie Boys 14U manager Jimmy Jackson. "We gave it a good shot."
In perhaps one of the most action-packed USCS Winged Outlaw Sprint Car Southern Thunder Series features ever at Lancaster Super Speedway, Eric Riggins Jr. worked his way past the field Saturday for the win.
When the green flag dropped, Morgan Turpen, who won at Carolina Speedway on Friday, roared into the lead with series point leader Terry Gray, Nick Snyder, Lance Moss and Riggins Jr. close behind.
Early on, Turpen, the "Queen of the Wings," stayed at the front of the pack until Snyder's car came to life.
AIKEN – The Lancaster Dixie Baseball Boys (14U) All Stars team stayed alive Monday in its quest for a state title with a 14-3 win over tournament host Aiken at Citizens Park.
Lancaster's 11-run win came after the local Dixie boys stars fell to Sumter, 14-4, for their first loss in the double-elimination field, as well as in all-star play.
Aiken broke out to a 3-0 lead in the top of the first, but Lancaster answered with a flurry and scored nine runs on seven hits to take a 9-3 lead after one inning.
I am so glad that the worst thing Indian Land residents are currently worrying about are their roads.
We here on the other end of the county located closer to Haile Gold Mine have our own concerns. The difference between ours and theirs lies in a much more all-encompassing, potentially harmful environmental impact, such as what Oceana Gold did to Brazil.
Oh, I know, they must not have all the regulations we have here. But what we do have are unconcerned politicians.
Elementary art teachers Melissa Caughman and Lisa Stamper decided in January as they were walking down Lancaster's Main Street to bring more art to town.
While brainstorming ideas, they named their new studio "a little art studio" with the intention to have their own storefront one day. Since January, Caughman and Stamper have been raising money through workshops and selling artwork to put toward a lease in downtown Lancaster. They're halfway there.
Teamwork and steady play sparked the winning team in the Lancaster Golf Club's par-three tournament July 23.
The winning foursome of Skip Small and Jason Blakley, along with the father-son duo of William Pate and Junior Pate, posted a 10-under 44 to notch a one-shot win.
"It was fun and quite different, very challenging," said Small, the team's A player.
Each team in the field included an A, B, C and D-level player.
Copyright © 2016
The Lancaster News is your source for local news, sports, events, and information in Lancaster County and Lancaster, South Carolina, and the surrounding area..


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I reinstalled v0.4 after the initial roadbumps got fixed. However since then I haven't actually needed to use the plugin to install any updates, etc. I tried installing a new plugin today and it kept saying my username and password were incorrect at the sftp screen. I new that wasn't the case so I reverted to v0.3. That same username and password worked fine with v0.3.
Not sure what changed, but I'll need to hang with v0.3 till this is fixed in v0.4. let me know if you'd like me to test anything regarding this issue.


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The 55-voice Augsburg Choir, under the direction of Peter Hendrickson, celebrates a long tradition of excellence in choral music. Praised for its high level of musicianship, the ensemble performs a diverse repertoire including compositions by well-known Scandinavian and American composers as well as a wide variety of music from throughout the world. The Augsburg Choir has premiered works by Knut Nystedt, Stephen Paulus, Steve Heitzeg, Carol Barnett, Egil Hovland, Kari Tikka, and Sigvald Tveit.
The Augsburg Choir's annual Advent Vespers services are regularly broadcast on public radio. In 2004, the 25th anniversary year, Twin Cities Public Television recorded and broadcast Advent Vespers, which was subsequently awarded a Regional Emmy Award. The Choir regularly tours locally, nationally, and internationally. In addition to national tours throughout the United States, the Choir has toured in Ireland, Sweden, Norway, Finland, Russia, Estonia, Hungary, the Czech Republic, Germany and China.
The Choir, in it's 82nd year, upholds the College's history of choral excellence dating back to the early part of this century. While honoring its roots in the Scandinavian Lutheran choral tradition, the Augsburg Choir has developed a generous and free choral sound, performing music from China to Colombia and South Africa to Russia. This choral tradition encompasses not only the mission of Augsburg but also the legacy of noted conductors such as Larry L. Fleming and Leland B. Sateren.
2015-2016 Performances:
Friday, October 9, 2015
Homecoming Convo: 10 a.m.
Hoversten Chapel, Augsburg College
Saturday, October 10, 2015
Homecoming Chapel: 10 a.m.
Hoversten Chapel, Augsburg College
Wednesday, October 21, 2015
Chapel: 10:40 a.m.
Hoversten Chapel, Augsburg College
Wednesday, November 18, 2015
Chapel: 10:40 a.m.
Hoversten Chapel, Augsburg College
Advent Vespers
Friday, December 4 at 5 & 8 p.m.
Saturday, December 5 at 2, 5, & 8 p.m.
Central Lutheran Church
Reservations Required
Sunday, February 8, 2015
8:30 & 9:45 a.m. Worship
St. Philip the Deacon Lutheran Church
17205 County Road 6, Plymouth, Minn.
Wednesday, March 2, 2016
Chapel: 10:40 a.m.
Hoversten Chapel, Augsburg College
Saturday, April 9, 2016
5 p.m. Mass
Basilica of Saint Mary
Corner of Hennepin Avenue & N. 17th Street, Minneapolis
Sunday, April 10, 2016 – 4 p.m.
Normandale Lutheran Church
6100 Normandale Road, Edina
Friday, April 29, 2016 – 8 p.m.
Home Concert – Augsburg Choir
Hoversten Chapel, Foss Center
Augsburg College
Saturday, April 30, 2016
8:30 a.m., Augsburg Baccalaureate
Hoversten Chapel, Augsburg College
May 2-11, 2016
The Augsburg Choir Tour


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|"Jersey Gods" begins in February '09
In New Jersey, a good man can be hard to find.
And while some ladies looking for the love of their lives may move their search across the Hudson River to Manhattan's social scene, the savvy Jersey girl at the heart of Image's February-launching "Jersey Gods" ongoing series broadens her search a little further to include cosmic depths of outer space, courtesy of writer Glen Brunswick and illustrator Dan McDaid.
"I want us to get into that [cosmic] world in a very accessible way," Brunswick ("The Gray Area," "Killing Girl") told CBR, explaining the fun of the series -- announced at this weekend's Baltimore Comic-Con -- is watching the culture clash between Jersey heroine Zoe and her war god hubby Barock's very separate lifestyles. "I think the idea is that Iâm giving a real modern sensibility to what Kirby did in the â70s and making it accessible and humanizing it by starting with this girl that hopefully everybody will relate to.
âWe follow the life of this girl that lives in New Jersey, and sheâs this high maintenance piece of work. Sheâs an amazing girl, but sheâs tough, and most of the guys she dates wonder if sheâs worth the effort that she puts them through. I think sheâs really kind of loveable in this slightly complicated mess sort of way, and sheâs a great window to center the book on. When we see the gods for the first time, itâs really through her eyes."
|Pages from "Jersey Gods"
When the gods crash through into our world, they select as their battleground a mall in Jersey suburb Cherry Hill, which happens to be the site of Zoe's latest broken relationship. As Barock pummels his way from shop to shop in combat with an evil god from his home planet of Nebulon, Zoe falls instantly for what she views as a perfect man who can live up to her high expectations. And as the old saying goes, first comes love, then comes marriage, then comes the crime fighting, family drama and otherworldly hilarity inherent in a cosmic god taking a bride in the 'burbs.
âWe see how they meet, and weâre going to get to see their courtship to a degree and what itâs like to marry a god from the perspective of the in-laws on earth and all the nonsense the gods have to do â the crazy gods that come to the wedding and turn everything on its ear,â said Brunswick, adding that the heart of the series will focus on life for the pair once they've settled in Cherry Hill full time. "Now, if [Barock] can just manage to survive his new in-laws, his household chores and his brideâs crazy social agenda, he might just carve out a moment or two to save the galaxy and protect the seedier parts of New Jersey."
Of course, while the earthbound adventures of a married couple comprise the driving force of "Jersey Gods," Brunswick assured readers that he's giving series artist Dan McDaid â a third place finalist in CBR's 2007 Comic Book Idol competition â plenty of opportunity to bring the slick, action-fueled visuals into the mix, as the history of Nebulon and its many gods play an equally important role.
|Pages from "Jersey Gods"
"10,000 years ago, [Barockâs] planet had this huge civil war, and his father was killed,â the writer explained. âAnd this war has affected all of the gods. They were like human beings kind of like us 10,000 years ago, but after this cataclysmic war, the survivors developed these godlike abilities. And itâs this war that changed them that the evil gods also refer back to when they think about their revenge. Barock lost his father, his brother was mortally wounded, but he did things â heroic things that his adversaries want to take revenge for."
Those story beats combine with Barock's own mysterious past to add a layer of mystery and excitement to the series over the long run. "Something has happened to his relationships, and heâs been a warrior for a very long time," Brunswick teased. "We donât know whatâs happened thatâs got him off to the idea of having a love relationship. We even see the difference between him and his best friend Helius who is a romantic lothario and is trying to coax him back into a relationship with a woman. Heâs a warrior, but there was a time he felt differently, and weâll learn more about that as the series goes along.
"This is a world like our own where there are no superheroes, where itâs like a first contact when the gods come down. The last time the gods came down, there was no media to document it, so the gods literally are the basis for all these earth myths that weâve had over the centuries. Theyâve lived for 10,000 years, but this is the first time that theyâll actually be documented."
|Pages from "Jersey Gods"
Aiding in the documentation of these gods will be a who's who lineup of A-list comics talent as the series first issues will include special alternate covers by the likes of Mike Allred, Darwyn Cooke, Paul Pope and Erik Larsen. Beyond that, Brunswick is inviting one of the industries biggest writers to contribute a back up serial that will run starting with "Jersey Gods" #2, Mark Waid. "I happened to show him this 'Jersey Gods' book, and he was really, really into it," Brunswick said. "Markâs going to do a backup story that involves the Nebulon civil war from 10,000 years ago. In each issue, rounding out the Jersey Gods universe will be this war story where we get to look into their past."
While Brunswick is also hoping to expand the backups in later issues with contributions from "Firestarter" scribe by Phil Hester, the main showcase of romance and action every month will be himself and McDaid. And with the new name on art chores, the writer isn't a bit worried about how things will turn out.
|Page from "Jersey Gods"
"I thought he was incredible and very polished. Heâs got a very Darwyn Cooke quality to his work,â Brunswick said. âI think that Darwyn is a big influence in Danâs work, and visually that really comes through. Iâm really, really excited about the way things worked out with Dan, and heâs really throwing himself into this. And not only is he a great storyteller, but he really does the thing that all great storytellers do in that he makes the characters more interesting by giving them bits of business. Even if theyâre not in the script, he puts these bits in, and as Iâm doing my rewrite once I get pages in, I can alter them to make them better and to fit his visuals. I feel that the really great artists do that. Back when I was working with John Romita, Jr. he was doing that kind of thing, and we got a chance to make the pages better. So Iâm really excited that Dan is this dazzling artist that at this point is pretty much undiscovered, but he wonât be for long."
Fans can discover "Jersey Gods" now in the form of two preview stories that ran in Image's recently released "PopGun 2" anthology before the first issue of the regular series hits just in time for February's New York Comic-Con.
|"Jersey Gods" art by Dan McDaid


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This has been a terrific plug-in so far. I was even able to figure out how to do a version of parent/child select lists without losing *all* my hair. :) Thanks for putting it together.
Is there an easy way to take the name of a field group and find the associated post ID so that I can look up what advanced custom fields are on that named group? For example, I have defined a set of fields for the post's related web site, title, subtitle and image. Going from the name "Related Information" how can I get to the list of fields post ID = 4? Ultimately I am trying to put together a custom field search page that lists options based on the custom fields on the post rather than a single generic search box.


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Essar to Spend $750 Million to Revamp Zimbabwe Iron and Steel
India's Essar Group took control of the Zimbabwe Iron and Steel Co. in a deal that will see the Zisco mill refurbished, said Welshman Ncube, the southern African nation's Industry Minister.
"In broad terms, the agreement provides for Essar to take over and release the Zimbabwean government from its debt obligations," Ncube said in Harare, the capital, today. "Essar will also complete all work relating to the re-lining of the blast furnaces and coke oven batteries," he said. Essar, through its Essar Africa Holdings Ltd. unit, will invest $750 million in the first phase of the project, Ncube added.
Zisco, which was state controled, hasn't operated since 2004 when it closed due to mounting debts and ageing equipment. Essar will have a 54 percent stake in the company, Ncube said.
Essar Africa's Vice Chairman Ravi Ruia said his company would have to first put in place "coal, uninterrupted electricity supplies, transport and logistics" in order to re-open the steel mill.
Zimbabwe President Robert Mugabe blocked the sale of Zisco to ArcelorMittal in June last year, saying the company was "too big" for the country.
Essar, which won a subsequent bid for Zisco, plans to produce steel within 12 to 15 months from now, Ruia said.
In a second phase, Essar will rehabilitate the rail link between Redcliff-based Zisco and the Hwange Colliery Co. in western Zimbabwe. Hwange is Zimbabwe's biggest digger of coal, which is needed to fire furnaces at Zisco's plant.
Essar will be the steelmaker's biggest shareholder, with government holding a minority 35 percent and the balance held by other shareholders, Ncube said.
"Zisco is totally insolvent, with debts in excess of $340 million. It has no value as a company and its debts exceed its assets," Ncube said, adding that most bidders valued the company at about $45 million during the due diligence process.


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503,427 deals (and counting)
from around the web, shared and ranked by a community of deal fiends like you.
Go to Deals.Woot.
Does your office allow the wearing of graphic t-shirts to work? Ours does! Hmm .. I wonder if no shirts is OK. Only one way to find out!
The ninja watercooler is back; I barely resisted the first time. And Night Shift....
Why resist temptation, when it's so much more fun to give in?
theco2 wrote:Why resist temptation, when it's so much more fun to give in?
True, but then shirtwoot! gets all my mad money...and I have to acquire new storage solutions...
Anyone else receive a small tshirt instead of the tote an item was supposed to be??
View All →


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Michael Schwendt wrote:
Yes, and I've successfully build using that repo baseurl before - I've not changed that entry. I'm getting the same error on fc3, fc4, fc5, and fcdevel mock builds. What are the correct repo url's I need to have?Executing /usr/sbin/mock-helper yum --installroot /var/lib/mock/fedora-5-i386-core/root groupinstall build build-minimal build-base http://newmirror.linux.duke.edu/pub/fedora/linux/core/5/i386/repodata/repomd.xml: [Errno 14] HTTP Error 404: Content-Length: 345Well, this repo baseurl is in invalid actually. Does it match your configuration files?
-- John


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Not Medical Advice: No, the white, oval pill with the imprint IP 465 is ibuprofen 600mg, not Vicodin. Ibuprofen is a pain reliever used to treat from various conditions such as headache, dental pain, menstrual cramps, muscle aches, or arthritis.
Possible side effects of ibuprofen are nausea, vomiting, headache, constipation, diarrhea, drowsiness or dizziness. If your condition persists or gets worse seek medical help right away. Daily use of alcohol and/or tobacco, especially when combined with this medication, may increase your risk for stomach bleeding.
According to a recent episode of The Dr. Oz Show, a large study revealed that two common over-the-counter pain relievers significantly increases the risk of deafness in women. Dr. Oz claims "Women who took ibuprofen or acetaminophen four to five times a week had a 12 percent increased risk in hearing loss..." The study Dr. Oz refers to is an article published in the American Journal of Epidemiology that studied 62,000 women between the ages of 31 and 48 over the span of 14 years.


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10 Superheroes Jason Bourne Could Escape From
Film, Comic Books
"The problem's plain to see: Too much technology!"
Continue Reading »
"Well he said, 'One thing, before I graduate / Never let your fear decide your fate.'"
Continue Reading »
In this feature, I spotlight five scenes/moments from within comic book stories that fit under a specific theme (basically, stuff that happens frequently in comics). Here is an archive of all the patterns we've spotlighted so far.
This week, based on a suggestion from reader Stephane S., we take a look at five instances where Captain America's unbreakable shield was broken…
Comics Should Be Good accepts review copies. Anything sent to us will (for better or for worse) end up reviewed on the blog. See where to send the review copies.


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Eliza sports a vintage bonnet from her mom's infant wardrobe
Eliza with her new friend Alida Myers
Eliza does a pretty good job holding her own bottle, although she sometimes struggles with proper bottle angle.
Eliza's first trip to Our Nation's Capital. Here she poses in Dupont Circle
With friends Josh & Beth Kvernon and their son Nathaniel
A mini Harvard House reunion atop a downtown DC apartment building. (Note Capitol dome to the left of Eliza's head)
Bath time for baby!
Eliza's first meal sitting on her high chair. So big!
Eliza thinks she is grown up because now she can suck on a real spoon, eat Cheerios, and is sprouting her second tooth.
Easter baby
Eliza's sleeptime acrobatics have increased greatly since we stopped swaddling her!
Eliza jumps
Eliza stands up (with help)


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After Record Sale, Another Richter Heads to Auction
German painter Gerhard Richter became the world's priciest living artist last week when his rainbow-colored "Abstract Painting" from 1994 sold for a record-setting $34.2 million at Sotheby's in London. Now, the auction house is hoping to capitalize on Richter's momentum by offering up another of the artist's abstracts from 1990, also called "Abstract Painting," in its Nov. 13 sale of contemporary art in New York. Asking price: $16 million.
Richter, who is age 80 and based in Cologne, has already created roughly 3,000 paintings over the course of his six-decade career, but collectors are paying a premium now for the series of large, colorful abstracts he painted using a squeegee between 1988 and 1994. From this series, only 65 paintings stand at least 6 feet tall—an appealing size for collectors seeking wall-power art—and a third of this tall group is already tucked into museums. That partly explains why collectors from Russian billionaire Roman Abramovich to dealer Larry Gagosian are jockeying to own the rest—42—still circulating in the marketplace.
More on the Art Market
Sale of Richter Painting Sets Record
Dealers say the anonymous American seller of this latest Richter at Sotheby's timed his move well: This particular abstract is over 6 feet tall, and its blue-gray palette was painted using heavy layers of paint in a style that's similar to the $15.1 million abstract bought by Mr. Abramovich during the market's last peak in 2008. By contrast, the current record-holder that Eric Clapton sold for $34.2 million last Friday was created with thinner paint layers and brighter colors.
Sotheby's also plans to test demand for the artist's family portraits on Nov. 13 by selling two paintings, both titled "IG," depicting Richter's second wife, artist Isa Gentzken. The house expects both to sell for at least $3 million apiece.
Write to Kelly Crow at email@example.com


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Information about your product
Thanks for choosing us in retirement - you're alongside more than one million people around the UK who also have their annuity with us.
Take a look at the articles below for more information on your annuity with PrudentialPrudential Annuities Customer Information Looking to 'sell' your annuity income?
For existing customers who currently have a plan with Prudential, and for customers who previously had a Stonehaven Lifetime Mortgage.
We no longer offer the Prudential Lifetime Mortgage product to new customers.
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We've been helping people with their investments for over 160 years and know how important it is to be financially stable.
Our range of investments are designed to help you build your financial foundation, whether you're saving over the medium to long term, or investing for your retirement.
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Preparing for the future is something that we all have to think about.Saving in a pension is a tax-efficient way of building up a fund to help provide for your retirement. And how you take your money when you get there is now more flexible than ever. Of course, as pensions are investment-based, there is the potential you may not get back what you put in.
Take a look at the articles below for more information on your pension with PrudentialAdditional Voluntary Contributions Section 32 Final Salary Pension (Defined Benefit) Prudential Personal Pension Money Purchase Plan Group Personal Pension Free Standing Additional Voluntary Contributions (FSAVC) Flexible Retirement Plan (with SIPP options) Executive Pension Plan Company Pension Transfer Plan (Bulk Section 32) Stakeholder Pension Self-Invested Personal Pension Retirement Annuity Plan
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18 Resort Reviews
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I'm longing to return now that they have been getting great snow for weeks in a row (The snow conditions weren't great when I visited soon after some rain, but the groomed trails were in decent shape and the glades and back country I was able to ski were firm but reeked of potential.) The views were amazing and the runs long and varied, although many had steep drops near the bottom- beginners are best off parking at the main lodge at the SUMMIT of this resort, which is on a bluff ("massif") overlooking the St Lawrence River. The terrain accessed from mid-stations and the upper summit quad seemed best for developing skiers. For the advanced skiers & riders, at the north edge of the resort there is an entrance to a hike-in back country area bounded between an in-bounds goaded trail and a long toboggan run that offers an easy escape route leading to the river lodge at the base. The hike is worth the fantastic glades that vary from tight to quite open, and with moderate to steep pitch. Modern detachable quads and an 8 person gondola serve all terrain.
I was very impressed by the local/healthy cafeteria foods; large meals were about $18 (US or CDN) including tax,and were delicious. No soda or fried foods available, which is fine by me. Nearby Quebec City was fantastic, too.
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Here is the site in question.
I am using a modified version of Foliage Mod by Derek Punsalan and this lightbox plugin.
All I need is basically for a way to have the images in the bottom portion of the screen behave in the same way as the images that live in the posts. I am not very experienced with Javascript at all. I've tried to call the same scripts listed on the header into "bottom.php" but no dice.
I've been back and forth with this for a while now and I can't think of what else to do. According to rhe plugin documentation, it scans the body of the page for images to assign the Javascript effects to them so it's obviously not scanning the bottom template for these even though it's still part of the body of the page. The links in the bottom are sctructured in the same way and have the same syntax as all other images on posts.
Any thoughts or advice would be greatly appreciated.