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The Thai Phase III clinical trial (RV144) showed modest efficacy in preventing HIV-1 acquisition. Plasma collected from HIV-1-uninfected trial participants completing all injections with ALVAC-HIV (vCP1521) prime and AIDSVAX B/E boost were tested for antibody responses against HIV-1 gp120 envelope (Env). Peptide microarray analysis from six HIV-1 subtypes and group M consensus showed that vaccination induced antibody responses to the second variable (V2) loop of gp120 of multiple subtypes. We further evaluated V2 responses by ELISA and surface plasmon resonance using cyclic (Cyc) and linear V2 loop peptides. Thirty-one of 32 vaccine recipients tested (97%) had antibody responses against Cyc V2 at 2 weeks postimmunization with a reciprocal geometric mean titer (GMT) of 1100 (range: 200–3200). The frequency of detecting plasma V2 antibodies declined to 19% at 28 weeks post-last injection (GMT: 110, range: 100–200). Antibody responses targeted the mid-region of the V2 loop that contains conserved epitopes and has the amino acid sequence KQKVHALFYKLDIVPI (HXB2 Numbering sequence 169–184). Valine at position 172 was critical for antibody binding. The frequency of V3 responses at 2 weeks postimmunization was modest (18/32, 56%) with a GMT of 185 (range: 100–800). In contrast, naturally infected HIV-1 individuals had a lower frequency of antibody responses to V2 (10/20, 50%; p=0.003) and a higher frequency of responses to V3 (19/20, 95%), with GMTs of 400 (range: 100–3200) and 3570 (range: 200–12,800), respectively. RV144 vaccination induced antibodies that targeted a region of the V2 loop that contains conserved epitopes. Early HIV-1 transmission events involve V2 loop interactions, raising the possibility that anti-V2 antibodies in RV144 may have contributed to viral inhibition.
Vaccination with the heterologous prime-boost vaccine regimen ALVAC-HIV (vCP1521) and AIDSVAXB/E (bivalent rgp120) was associated with a 31.2% reduction in HIV-1 acquisition at 42 months in the modified intent to treat population and provided the first opportunity to study the immune correlates of HIV-1 vaccine-induced protection in humans.1–3 Evaluation of the vaccine regimen in Phase I/II studies showed that this vaccine induced both cellular and humoral immune responses.4,5 Immunogenicity studies have demonstrated the induction of neutralizing antibodies against tier 1 strains of HIV-1, antibody-dependent cell-mediated cytotoxicity (ADCC), antigen-specific lymphoproliferative responses, and CD8+ cytotoxic T lymphocyte (CTL) responses.4–9 Vaccination with recombinant Env proteins alone in AIDSVAX B/E (Vax003 trial) and AIDSVAX B/B (Vax004 trial) generated strong humoral immune responses but did not decrease HIV-1 acquisition in Thai injecting drug users or North American men who have sex with men, respectively.10–12
The role of antibodies in preventing HIV-1 acquisition has been suggested by passive13–16 and active immunization studies in nonhuman primates17,18 as well as by preliminary data in mother–infant transmission studies,19 but has not been confirmed as a general principle of protection in humans. Recently it was found that antibodies against subtype B gp120 V1V2 loops appeared to correlate with decreased risk of infection in RV14420; however, broadly neutralizing antibodies were not seen in that study,21 and neither neutralizing antibodies nor antibody-directed cell-mediated cytotoxicity appeared to correlate with infection risk.20 Nonetheless, several regions of the HIV-1 gp120 glycoprotein contain epitopes that induce antibodies that may play a role in protection. Epitopes found in the second variable (V2) loop have been associated with HIV-1 Env functions that are important in infectivity, CD4 receptor and CCR5/CXCR4 coreceptor usage, escape from antibody neutralization, the formation of Env trimers, and the protection of the coreceptor binding site.22–28 Sequence variation within the V1/V2 loops alters neutralization resistance and viral escape is associated with V2 loop mutations.29–33 Compared to viruses circulating in chronically infected persons, transmitted viruses (subtypes A and C) appear to have shorter V1/V2 regions and fewer N-linked glycosylation sites.30,34–36
The V2 loop contains a putative α4β7 integrin binding motif (LDI/V) at amino acid (aa) residues 179–181 (HBX2 numbering system) that was shown to interact with the gut mucosal homing receptor.23,24 Under physiological conditions, the α4β7 receptor facilitates the migration of lymphocytes to the gut and some have postulated that HIV-1 exploits this interaction to infect lymphocytes localized at mucosal tissues.23,24,37,38 However, further studies are needed to confirm the role of α4β7 in viral transmission with unliganded gp120 trimers.39
Broadly neutralizing antibodies that interact with quaternary neutralizing epitopes (QNE) of trimeric native Env spikes have been shown to interact with conserved aa embedded within the variable (V) loops of gp120 including the V2 loop.40–47 Furthermore, antibodies found in individuals that exhibit potent cross-clade neutralizing activity have been shown to target conserved regions of the V1, V2, and V3 loops, providing evidence that V2 contributes to the formation of QNE that can induce potent cross-clade neutralizing antibodies.25,43 The structure of scaffolded V1/V2 complexed with the broadly neutralizing monoclonal antibody PG9 has been resolved.47 This potent antibody binds to glycans and aa sequences within the V1/V2 regions. Antibodies against the V2 loop target several conserved residues that are sensitive to glycosylation patterns and changes within the loop alter the efficiency by which antibodies neutralize HIV-1.26,30,41,48
T cell peptide epitope mapping studies with peripheral blood mononuclear cells (PBMCs) from HIV-1-uninfected RV144 vaccine recipients using the interferon-gamma ELISpot assay demonstrated preferential CD4+ T cell responses to the V2 loop region compared to other regions of gp120.49 Antibody epitope mapping studies were undertaken to assess whether there was a similar preferential targeting of regions within gp120.
In this study we report that the RV144 vaccine regimen generated antibodies to the V2 region of gp120. Using cyclic as well as linear peptides, we demonstrate that the vaccine-induced antibodies recognize the mid-region of the V2 loop. A role for these antibodies in protection from HIV acquisition was recently suggested by a genetic sieve analysis of subjects who enrolled in RV144 and became HIV-1 infected.50
The design and findings of the RV144 Phase III clinical trial have been published previously.1 In this study we used three sets of RV144 plasma samples (sets A, C, and Z) that were randomly generated by the Statistical Center for HIV/AIDS Research Prevention (SCHARP), Seattle, WA, and the EMMES Corporation, Rockville, MD from volunteers who received all immunizations according to schedule and were HIV seronegative at the end of the study (Table 1). It was not possible to use the same sample set for all studies due to the limited volumes of sample available. However, all plasma separations were performed at the same centralized laboratory. Each plasma set consisted of uninfected subjects in the per-protocol population with ratios of 4:1 vaccine:placebo for both male and female subjects. Simple random sampling was used within each of the four strata. For peptide microarray and biotinylated linear peptide analyses, plasma samples from 80 vaccine and 20 placebo recipients, designated plasma set A and set C, were used, respectively (Table 1). For cyclic (Cyc) peptide analysis, plasma samples from 32 vaccine and 8 placebo recipients, designated plasma set Z, were used (Table 1). RV144 plasma samples from three different time points were tested: baseline or prevaccination (visit 1), 2 weeks (visit 8), and 28 weeks (visit 9) post-final inoculation. The RV144 trial was conducted under a protocol approved by IRBs from the Thai Ministry of Public Health, Mahidol University, the Scientific Subcommittee of the National AIDS Committee, Thailand, the Royal Thai Army, Thailand, and the U.S. Army Medical Research and Materiel Command, United States (http://clinicaltrials.gov number NTC00223080).
Plasma samples from Thai blood donors infected with HIV-1 circulating recombinant form (CRF) 01_AE (WRAIR protocol #1617) were used in ELISA and Biacore assays with Cyc V2 peptides.
CHO-expressed recombinant gp120 proteins derived from MN and A244 were kindly provided by Dr. Marc Gurwith, Global Solutions for Infectious Disease, San Francisco, CA. HIV-1 CRF01_AE 92TH023 gp120 protein was expressed in 293T cells and purified using Galanthus nivalis lectin columns.
Peptide microarrays and Cyc peptides were synthesized by JPT Peptide Technologies. Peptides were cyclized by disulfide bond formation (Fig. 1A) and the purity was determined to be greater than 90% by high-pressure liquid chromatography and mass spectrometry. The aa sequences of Cyc V2 and V3 peptides were based on vCP1521 Env glycoprotein of HIV-1 CRF01 AE (92TH023 strain) GenBank accession number EF553537.1 (Fig. 1). Cyc V2 peptides of varying lengths as well as those with scrambled mid-region (Scr MR) or scrambled flanking regions (Scr Fl) were synthesized with or without biotin at the amino terminus of the peptide. HIV-1 strains 92TH023 and A244 have identical V2 loop mid-regions (Fig. 1B). Cyc V3 peptide was not biotinylated. Cyclic nonbiotinylated peptides were used in all ELISAs and biotinylated peptides were used for all Biacore binding studies. The sequences of Cyc peptides are shown in Table 2. The aa sequences of the scrambled regions of the mid-region of Cyc V2 Scr MR and the flanking regions of Cyc V2 Scr Fl are shown in bold (Table 2). The integrin binding motif, LDI, is underlined.
Peptides representing the more sequence-conserved segment in the first two-thirds of the V2 loop were selected from the set of all recorded V2 sequences in the Los Alamos National Laboratory (LANL) database in order to maximize predicted functional diversity. The sequences of the linear V2 peptides are shown in Table 2, and the integrin binding motif in each of the linear peptides is underlined. Polar and charged amino acids mediate most chemical functions of proteins such as solvent interactions (surface accessibility), binding, posttranslational modification, and catalysis. Peptide 1 was selected as the V2 loop mid-region from a strain with the most charged and polar aa among circulating strains from subtype A, strain QB585.2102M.Ev1v5.C. Peptide 3 was selected as the V2 loop mid-region sequence found in circulating strains with the most polar and charged aa and also exhibiting the most common V2 length of 39 aa from subtype A, strain 01TZA341. Peptide 2 represents the sequence most commonly found in circulating subtype B strains recorded in the LANL database as of May 2011, strain 878v3_2475. Peptide 4 represents the consensus sequences for V2 in the region covered by peptides 1–3. Peptide 6 contains the N-terminal 14 aa of peptide 1. Peptides were biotinylated at the N-terminus during synthesis and were purchased from Genemed Biotechnologies, Inc., South San Francisco, CA.
Carboxymethylcellulose 5 (CM5) chips, streptavidin (SA) chips, and the amine coupling kit were purchased from GE Healthcare, Piscataway, NJ. Lysozyme, Costar Spin-X centrifuge tube cellulose acetate filters (0.22μm pore size), Dulbecco's phosphate-buffered saline (D-PBS), Tween 20, and Thimerosal were purchased from Sigma-Aldrich, St. Louis, MO. Affinity purified sheep antihuman IgG (gamma chain specific) antibodies were purchased from The Binding Site, Birmingham, UK. Skim milk was from Applichem, St. Louis, MO. ABTS peroxidase substrate solution was from KPL, Gaithersburg, MD. Horseradish peroxidase-conjugated goat antihuman IgG antibodies were from Bethyl Laboratories, Montgomery, TX. Strepta Well plates were from Roche, Mannheim, Germany. Alkaline phosphatase-conjugated goat antihuman IgG was purchased from SouthernBiotech, Birmingham, AL. Anti-IgG Cy5 antibody was from Jackson ImmunoResearch Laboratories, West Grove, PA. Superblock T20 PBS blocking buffer, Immulon 2HB U bottom, and Immulon 4HBX flat ELISA plates were purchased from Thermo Fisher Scientific, Rockford, IL.
Each array consisted of 1423 tiled Env peptides arranged in three identical subarrays, allowing for triplicate evaluation of the entire set of peptides. Peptide sequences covered the entire gp160 subtype consensus sequences from six HIV-1 group M subtypes (A, B, C, D, CRF01, and CRF02) and a consensus group M gp160, Con-S,51 for a total of 1423 peptides (15-mers overlapping by 12 aa). We used consensus sequences rather than the vaccine strains, as we wished to focus on cross-reactive responses rather than type-specific responses to the vaccine immunogens. The peptides were generated using an alignment of the seven consensus sequences with LANLs PeptGen tool (www.hiv.lanl.gov), so that peptides remain in register throughout the Env despite insertions and deletions, and identical peptides found in more than one subtype were represented only once. Microarray development was performed using the HS4800 Pro Hybridization Station (Tecan, Männedorf, Switzerland). All arrays were blocked with Superblock T20 PBS blocking buffer for 0.5h at 30°C, followed by a 2-h incubation at 30°C with heat-inactivated plasma diluted 1:100 in Superblock T20. Arrays were then incubated for 45min at 30°C with a secondary antibody, 1.5mg/ml anti-IgG Cy5 antibody diluted 1:1000 with Superblock T20. Washes between all steps were with PBS containing 0.1% Tween. Arrays were scanned at a wavelength of 635nm using an Axon Genepix 4300 Scanner (Molecular Devices, Sunnyvale, CA) at a PMT setting of 600, 50% laser power. Images were analyzed using Genepix Pro 7 software (Molecular Devices). Spot intensities were corrected for prevaccine results.
Peptide microarray spot intensities were extracted from the raw images using the GenePix software, and peptide intensities were calculated as follows: (1) Spot intensities were defined as foreground median intensity minus background median intensity. (2) Resulting intensities were log2 transformed and centered by subtracting the average (log2) intensities of the empty (control) spots on the slide. (3) For each unique peptide, a single intensity was defined as the median of the triplicate intensities on the slide for that peptide. To remove effects unrelated to true peptide-binding activity, such as nonspecific binding of primary antibody in the sample, array normalization was performed in an attempt to remove such effects. Nonspecific binding may be related to physicochemical properties of the peptide, we used the z-scales to correct for nonspecific binding biases.52 To arrive at a summary z-scale score for each peptide, the z-scale values of the individual aa comprising that peptide were summed. Here, the normalization was done in two stages: (1) a regression model for the probe intensities was derived from their overall z-scores and (2) each probe was normalized by subtracting its predicted intensity (representing the bias) from the observed intensity. A linear model with t4 distributed errors was used, which provides more robustness against probe outliers as described previously.53 This normalization was performed on each array separately. Once the data were normalized, the probe intensities were smoothed and a score was calculated for each peptide that was used to detect binding regions. Based on the assumption that an antibody binds a region of approximately 9 aa or more, normalized peptide intensities were smoothed using a sliding window mean statistics of size 9 aa, i.e., peptides with HXB2 positions within 9 aa of one another were grouped together. This smoothing step can be made subtype specific (e.g., for subtype A) or across all subtypes to borrow strength across multiple peptides. For baseline correction, samples were corrected by subtracting, for each sample, the peptide intensities measured prevaccination for that sample.
Once scores have been calculated and corrected for baseline, the score cutoff to call positive peptides can be set arbitrarily, determined based on simple validation, or based on a false detection rate (FDR) cutoff. Here the placebo group from the RV144 data was used to estimate the FDR and set at an appropriate threshold. Peptides with log2-fold change greater than 1.1 (with respect to baseline) were called positive, which led to an FDR of approximately 5%.
Briefly, U-bottom 2HB plates were coated with either 1μg/ml of cyclic peptide or with 3μg/ml of recombinant gp120 in D-PBS at 4°C overnight. Wells were washed three times with wash buffer (PBS, 0.1% Tween 20, and 0.01% Thimerosal, pH 7.4) using Microplate Washer ELX405, Bio Tek, Winooski, VT, and blocked with blocking buffer (D-PBS, 5% skim milk) for 2h at room temperature. Plasma was initially diluted 1:100 in blocking buffer and then serial 2-fold dilutions were performed and added to wells for 2h at room temperature. Wells were washed five times with wash buffer and HRP-conjugated goat antihuman IgG (1:25,000) was added to wells for 1h at room temperature. Plates were washed five times with wash buffer, 100μl/well of substrate was added, and color was allowed to develop at room temperature for 1h in the dark. Plates were read at A405nm using an ELISA reader Spectramax 340 PC, Molecular Devices. The data are expressed as end point titers, with the titers being defined as the reciprocal of the highest dilution that yielded an absorbance value above 2.5 times the background value (wells that did not contain recombinant protein or peptides).
Briefly, StreptaWell plates (Roche) were coated with 1μg/ml biotinylated linear V2 peptides for 1.5h at 37°C and then washed six times with PBS containing 0.05% Tween-20, pH 7.4. Plates were incubated for 1.5h at 37°C with RV144 plasma diluted 1:100 in RPMI media containing 15% fetal bovine serum. The plates were washed six times and alkaline phosphatase-conjugated goat antihuman IgG (1:2,000) was added for 1.5h at 37°C. After washing, 10% diethanolamine substrate was added for 30min to develop color, and the plates were read at A405nm. All reagents were added in a volume of 50μl/well and each sample was run in duplicate.
ELISA antibody titers were calculated as the reciprocal plasma dilution using serial 2-fold dilutions of plasma from 1:100 to 1:12,800 and are expressed as the reciprocal end-point titer. Geometric mean titers (GMT) were calculated from the reciprocal end-point titers. Data analyses and graphs were generated using Graphpad Prism version 5. Statistical comparisons were made using nonparametric tests and a two-sided p-value of <0.05 was considered significant.
To confirm the findings of the ELISA studies, surface plasmon resonance (SPR) measurements were conducted with a Biacore T100 using CM5 or SA chips prepared as described below. Lysozyme was immobilized onto a CM5 chip using the amine coupling kit. Unbound free amines were quenched with ethanolamine. All solutions used during the immobilization steps had a flow rate of 10μl/min and all experiments were performed at 25°C. The immobilization wizard packaged within the T100 control software was used to immobilize 500nM lysozyme in 20mM sodium phosphate, pH 7.4 (5min contact time) in flow cell 1. Cyc V2 biotinylated peptides were prepared at 0.1–1μM in Tris-buffered saline, pH 7.4, and allowed to flow for 2–10min over the streptavidin-coated surface of the SA chip, which resulted in immobilization of approximately 895, 1,401, and 2,073 response units (RU), respectively, for the 16, 25, and 42 aa containing Cyc V2 peptides. Cyc V2 Scr MR (1900 RU) and Cyc V2 Scr Fl peptides (1,850 RU) were immobilized to the respective flow cells.
Plasma samples were heat inactivated (56°C, 45min), centrifuged, and the supernatant filtered prior to use. The plasma was diluted 1:50 in Tris-buffered saline, pH 7.4. The diluted plasma samples were passed over the chip surface at 30μl/min for 3min followed by a 5-min dissociation period. At the end of the 5-min period, a 50nM solution of affinity-purified gamma chain-specific sheep antihuman IgG antibody was passed over the peptide-coated Ig-bound surface for 2min at a flow rate of 10μl/min. After a 70-s dissociation period, the biotinylated peptides were regenerated using a 30-s pulse of 50mM HCl, a 30-s pulse of 100mM EDTA in 20mM Tris, pH 7.4, another 30-s pulse of 50mM HCl, followed by a 1min injection of Tris-buffered saline, pH 7.4. Nonspecific binding was subtracted and data analysis was performed using the BIA evaluation 4.1 software. The reported RUs for the IgG-specific values are the difference between the average value of a 5-s window taken 60s after the end of the anti-IgG injection and the average value of a 5-s window taken 10s before the beginning of the anti-IgG injection. The data (RU) are presented as dot plots for individual plasma samples. A representative experiment of three independent experiments is shown.
To investigate the antibody responses induced by the RV144 vaccine regimen, we performed ELISAs using the vaccine matched recombinant gp120 proteins from the three HIV-1 vaccine strains, MN, 92TH023, and A244. A244 and MN comprised the protein boost and 92TH023 was in the canarypox vector. Set Z plasma samples from visit 1 (prevaccination), visit 8 (two weeks), and visit 9 (28 weeks) post-last inoculation were tested against gp120 proteins (Fig. 2). The visit 1 responses were negative except for three vaccinees' samples where the responses were above the cutoff value for the assay (Fig. 2A). However, the visit 1 responses for these three samples were markedly lower than visit 8 and 9 samples and were considered nonspecific. Of the 32 vaccinees, 31 (97%) had robust antibody responses against all three recombinant proteins at visit 8 with reactivity against MN and A244 being the highest followed by 92TH023 (Fig. 2A). One of the eight placebo subjects had antibody responses to both 92TH023 and A244 proteins at all three visits but no responses to MN (Fig. 2B). Although the frequency of positive responses at visit 9 did not change 6 months after administration of the last vaccination, the magnitude of antibody responses against all three gp120 proteins declined with antibody levels to 92TH023 being the lowest followed by A244 and MN (Fig. 2A).
Binding antibodies in RV144 set A plasma samples were assessed by peptide microarray analysis using linear overlapping peptides covering the entire V2 loop of all major genetic subtypes and CRFs of gp120 (Fig. 3). Of the seven sets of HIV-1 gp120 peptides tested, six displayed antibody responses against V2. Notably, there was little or no reactivity with the V2 peptides of subtype B despite the fact that the vaccinees had received gp120 protein boosts derived from both subtypes B and CRF01_AE (Fig. 3A). Based on this analysis, the RV144 vaccine induced antibody responses that specifically targeted 15-mer peptides covering the V2 loop with 25% (20/80) of the vaccinated subjects having antibodies to this region (Fig. 3B).
Based on the antibody responses to linear peptides that span the V2 loop in peptide microarray analysis, we concluded that antibodies interacted with the mid-region of the V2 loop having the following aa sequence—KQKVHALFYKLDIVPI—and corresponding to HXB2 aa 169–184 (Fig. 1). The mid-region of the V2 loop contains highly conserved aa including the α4β7 integrin binding motif LDI/V23 and ALFY, but also a less conserved region (KQKVH). To investigate further the induction of V2 loop-specific antibodies in RV144, we synthesized Cyc V2 peptides based on the canarypox Env glycoprotein 92TH023 (see Materials and Methods, Table 2, and Fig. 1). 92TH023 and A244 V2 loops vary by two aa at positions 188 and 189 (HXB2 numbering) but the mid-region is identical. The Cyc V2 peptide contained 42 aa extending from aa 158 to 199 (corresponding to HIV-1 HXB2 aa 157–196). To analyze the antibody responses targeting the flanks and mid-region of the V2 loop, two additional Cyc peptides were used (Table 2).
Plasma samples from visits 1 and 8 were tested against the Cyc V2 peptides by ELISA (Fig. 4). No antibody responses to Cyc V2 were detected in prevaccinated (visit 1) plasma. Of the 32 vaccinated subjects tested at visit 8, 31 had positive IgG antibody responses against Cyc V2 (97%) at visit 8 (Fig. 4A). Antibody responses to the Cyc V2 Scr Fl peptide were almost identical to those against the Cyc V2 peptide. Unlike Cyc V2 and Cyc V2 Scr Fl peptides, the IgG antibody responses against Cyc V2 Scr MR were significantly reduced with only 16/32 (50%) plasma samples being above background values (p<0.001) indicating that the majority of antibody responses against V2 targeted the mid-region of the loop (Fig. 4A). Identical responses were seen with Biacore for all three peptides (data not shown).
To better characterize the antibody-binding epitope within the Cyc V2 peptide, two shorter Cyc V2 peptides, Cyc V2 (25aa) and Cyc V2 (16aa), were synthesized and tested by Biacore analysis (Table 2 and Fig. 4C). There was a single positive responder among the vaccinees at visit 1 to the Cyc V2 (25aa) peptide (Fig. 4C). Both low-frequency and weak binding were observed with the Cyc V2 (25aa) peptide (5/32, 16%; p<0.001) but no binding was observed with Cyc V2 (16aa) (Fig. 4C). No antibody responses were detected in plasma samples from visits 1 and 8 of placebo recipients to any of the Cyc V2 peptides (Fig. 4C).
To determine the durability of antibody responses against the V2 loop, the antibody responses at visits 8 and 9 were compared using the Cyc V2 peptide. The frequency of antibody responses at visit 8 was high 31/32 (97%), with a GMT of 1,100 and a range of 200–3,200 (Fig. 5A). The frequency and magnitude of antibody responses decreased significantly at visit 9 (6/32, 19%) compared to visit 8 (p<0.001) with a GMT of 110 and a range of 100–200 (p<0.001). Biacore analysis using the same peptide and plasma samples showed similar results with antibody responses to V2 diminishing by visit 9 (Fig. 5B). Reactivity was not detected in placebo recipients or vaccinees prior to immunization by either ELISA or Biacore assays (Fig. 5A and B).
Antibody responses to V2 were also examined using five linear biotinylated V2 loop-derived peptides that are heterologous in sequence to the RV144 immunogens (Table 2 and Fig. 6). In this ELISA, RV144 plasma samples from set C were used (80 vaccine and 20 placebo recipients). Antibody responses against all peptides were detected at visit 8 with peptide 6 having the highest frequency (93%) of responses followed by peptides 1 (84%), 3 (68%), 4 (39%), and 2 (38%), respectively (Fig. 6A). Poor responses against linear peptides 4, 6, and consensus B V2 peptides (microarray analysis) indicated that the presence of valine at position 172 is important for antibody binding. The responses to linear peptides declined to background levels at visit 9 and were similar to those observed with the V2 cyclic peptide using ELISA and Biacore (Fig. 5). Two samples from placebo recipients reacted with the linear peptides 1 and 6 and one sample reacted with peptide 2 (Fig. 6B). The sample reacting with peptides 1 and 6 (shorter version of peptide 1) was not the same as with peptide 2. These responses were considered nonspecific.
V2-specific IgG antibody responses generated by the RV144 vaccine regimen were compared to those induced by HIV-1 infection. Antibodies to Cyc V2, Cyc V2 Scr MR, and Cyc V2 Scr Fl were measured using plasma samples from 20 HIV-1 CRF01_AE-infected individuals and four HIV-1-uninfected Thai subjects. Ten of the 20 infected subjects (50%) had antibodies to Cyc V2 (GMT: 400; range: 100–3,200) (Fig. 4B), which were significantly lower than the frequency and magnitude for the RV144 vaccine group (p<0.001). Similar antibody-binding responses in frequency and magnitude were measured with the Cyc V2 Scr Fl peptide suggesting that the antibody responses in HIV-1 infection also target the mid-region of V2 loop and are similar to those generated by the RV144 vaccine regimen (Figs. 4A and and5).5). The magnitude of antibody responses to the Cyc V2 Scr MR peptide was low, 30% (6/20, Fig. 4B), and antibody responses were not detected in HIV-1-uninfected controls (Fig. 4B).
Biacore analysis using plasma samples from 18 of the 20 HIV-1-infected subjects tested by ELISA gave similar results (data not shown).
Antibody responses to V2 were also compared to those generated against the V3 loop. Plasma from 32 vaccinated and eight placebo recipients at visits 1 and 8 were tested against Cyc V3 (Table 2 and Fig. 7). The frequency (18/32, 56%) and magnitude (GMT: 200; range: 100–800) of antibody responses against V3 were lower in vaccinated subjects (Fig. 7). In comparison, the Cyc V3 antibody responses in HIV-infected individuals were both higher in frequency (19/20, 95%) and magnitude (GMT: 3,600; range: 200–12,800) (Fig. 7). Antibody reactivity against the Cyc V3 peptide was not detected in uninfected control subjects or vaccine recipients at visit 1. Unlike HIV-1-uninfected RV144-vaccinated subjects, HIV-1-infected individuals had a higher frequency of antibody responses against 92TH023 V3 (95%) compared to V2 (50%; p=0.003). Antibody responses against MN Cyc V3 were not tested.
The levels of Cyc V2 and V3 antibody responses were compared in the same HIV-1-infected individuals. Seven out of 20 (35%) subjects tested had Cyc V3 loop antibody responses with a GMT of 4,800 (range: 1,600–12,800) but undetectable levels of antibodies against the Cyc V2 loop (Fig. 8A). In 3/20 (15%) subjects, the V2 response was high (GMT: 3,200) but still below the V3 antibody responses (GMT: 5,100; range: 3,200–6,400). All HIV-1-infected subjects had higher responses to Cyc V3 than to Cyc V2 (Fig. 8A). Of the 20 infected subjects, one (5%) did not have an antibody response to either V2 or V3 loop peptides.
Of the RV144 vaccinated subjects, 31/32 (97%) had an antibody response to the Cyc V2 peptide, but only 18/32 (56%) had antibody responses to the Cyc V3 peptide (Fig. 8B). None of the 32 vaccinated subjects had a V3 loop antibody response that was higher than that of V2 (Fig. 8B). One of the vaccinated individuals did not have antibody responses to either the V2 or V3 loops.
The estimated efficacy in the RV144 vaccine trial was 31.2% by 42 months in the modified intent to treat population,1 but data through months 12 and 18 showed estimated efficacy of 60% and 44%, respectively.54 Initial studies demonstrated that binding antibody to vaccine gp120 antigens was found in 95% of vaccine recipients 2 weeks post-final vaccination but waned substantially over the next 6 months. Using peptide microarray analysis, ELISA, and Biacore studies we demonstrated that the immunogens used in the RV144 trial induced antibodies to the V2 loop of gp120 from diverse HIV-1 subtypes. These responses were high in frequency and magnitude but also declined rapidly to nearly undetectable levels at 28 weeks post-last vaccination. Haynes et al.20 have shown that IgG against a conformational V1/V2 epitope from 2 weeks postvaccination was inversely correlated with infection rate in the RV144 trial. While diminution of the V2 antibody may have corresponded to waning vaccine efficacy in RV 144, a mechanistic link between the two observations is purely speculative at this point. Further research in the "time-dependent" correlates analysis will be required to pursue this hypothesis.
Using scrambled cyclic and linear peptides of the V2 loop we determined that antibodies induced by RV144 target the mid-region of the loop, which includes highly conserved aa including the integrin-binding motif LDI/V but also less conserved regions. It is likely that RV144 antibodies have multiple binding epitopes within the mid-region and further studies are needed to characterize their binding specificities.55 Two monoclonal antibodies isolated from RV144, CH58, and CH59 have different but partly overlapping binding.56 It is likely that not all antibody responses targeting the V2 loop are protective and certain epitopes could be more protective than others in preventing HIV-1 acquisition. Furthermore, volunteers who develop antibodies to Cyc V2 may or may not develop antibodies to gp70, a scaffold glycosylated protein containing the V1/V2 loops used in the case control study.20 Scrambling the flanks of the V2 loop had minimal effect on the frequency or magnitude of binding in vaccinated and infected subjects indicating that the majority of the antibodies target epitopes within the mid-region. To further define the binding epitopes of these antibodies, Cyc V2 (16aa) and Cyc V2 (25aa) peptides were used. Of these two peptides, weak responses were observed only to Cyc V2 (25aa). Peptide Cyc V2 (25aa) includes all aa found in the V2 mid-region except KQKV at the N terminus of the peptide, suggesting that these four aa could be important for epitope recognition by V2 antibodies. Alternatively, conformational changes of shorter cyclized peptides may not bind to antibodies. It is possible that certain conformational antibodies induced by the recombinant proteins used in the vaccine regimen may not bind to peptides. Furthermore, antibodies targeting glycosylated epitopes within the V1/V2 loops were not investigated in this study.
Recently, a proposed structure of scaffolded V1/V2 loops complexed with the broadly neutralizing monoclonal antibody PG9 has been reported.47 V1/V2 loops form an intertwined four-stranded β sheet domain composed of four antiparallel β-strands designated A, B, C, and D. PG9 interacts with N-linked glycans on the V1/V2 loop but also interacts with aa within strand C, which is part of the V2 loop region. Strand C is part of the mid-region of the V2 loop and interacts strongly with antibodies induced by the RV144 vaccine regimen. In addition to its interaction with PG9, strand C contributes to the formation of a nonglycosylated hydrophobic core and participates with the other three strands to form a single topological entity.47 The consequence of interaction of this topological entity with antibodies is currently unknown.
Comparing antibody responses and sequence alignments of peptides used in the study, we conclude that valine at position 172 (HXB2 numbering) could be important in epitope formation and reactivity. Valine at this position is characteristic of CRF01_AE V2 loops and differs from the glutamate found most commonly at this position in subtype B strains. Linear and cyclic peptides lacking valine at position 172 were poorly reactive or nonreactive with the samples tested.
The generation of antibody responses against the mid-region of the V2 loop was also confirmed by linear biotinylated peptides. Of the five peptides tested, peptide 6 had the highest response followed by peptides 1 and 3. Peptide 6 is a shorter version of peptide 1 and lacks the LDI motif. The high antibody response to this peptide suggests that the core epitope recognized by antibodies to linear peptides does not include LDI; however, this does not imply that either the LDI is not targeted by antibodies to conformational epitopes in V2 or is not sterically blocked by V2 antibodies to the core epitope. In general, linear peptides had weaker responses than the 92TH023 Cyc V2 peptide possibly due to the lack of high homology with the aa sequences of the V2 loop used in the vaccine and to conformational differences as cyclic peptides may assume a more complex conformational folding.57–61
Using microarray analysis, RV144 vaccinee plasmas were reactive with six of the seven sets of overlapping V2 peptides from consensus gp160 proteins from group M and subtypes A, C, D, CRF01, and CRF02; only those from subtype B were negative. The frequency of antibody responses to the V2 loop by microarray peptide analysis was much lower than that observed for cyclic V2 peptides using ELISAs and Biacore. The difference in binding could be attributed to aa sequence variability but also to conformation, adsorption, or peptide length.
In contrast to the V2-specific IgG antibody responses induced by the RV144 vaccine regimen, the Cyc V3 loop responses in RV144 were of lower magnitude (GMT of 200) and frequency (56%), thus indicating that RV144 did not induce vigorous antibody responses to the clade E V3 loop region. However, responses to the cyclic V3 loop in HIV-1-infected individuals were higher in magnitude (GMT of 3,600) and frequency (95%). Infected subjects had robust antibody responses to the Cyc V3, but 35% of those having strong antibody responses to V3 lacked antibodies to the Cyc V2. There was an 18-fold higher response to the Cyc V3 peptide in HIV-1-infected individuals compared to the RV144 vaccinees. However, V2 loop-specific antibodies in infected individuals were much less frequent (50%) and of lower magnitude than those induced by the RV144 vaccine regimen. Whether this observation of high V2 and low V3 responses in vaccinees compared to HIV-1-infected subjects is related to the vaccine antigen is unknown. Vaccine-induced antibody responses have many similarities to antibodies found in infected individuals but generally are not neutralizing in cell assays.62
Heterosexual transmission is usually a single infectious event and productive infection reflects the expansion of the founder virus.36,63,64 The V2 loop has a number of features that might involve initial events in infection. It has a highly conserved length among early or transmitted/founder isolates and length and glycosylation of the V2 loop are commonly associated with escape from neutralizing antibodies.30,35,65 Furthermore, V2 contains a putative α4β7 integrin-binding motif of uncertain significance.39 Although V2 has been implicated in early virus transmission the precise mechanisms of attachment and cell-to-cell dissemination are not well defined.23,24,36,65–67 Rolland et al. have reported that sieve mutations in HIV-1 breakthrough infections in RV144 contain mutations at positions 169 and 181 in V2.50 T cell epitope mapping studies using PBMCs from HIV-1-uninfected RV144 vaccine recipients and the interferon-gamma ELISpot assay also showed preferential targeting of the V2 loop and that the responding cells were CD4+.49 Taken together with the potential role of antibody to V2 as a correlate of protection,17,20 these findings suggest that the RV144 vaccine regimen may have induced protection by targeting the V1/V2 loops.
The generation of humoral immune responses by ALVAC-HIV and AIDSVAX B/E to the mid-region of the V2 loop raises hypotheses to test in regard to correlates of protection against HIV-1 infection. The V2 loop (like the V3 loop) contains many conserved features, and several anti-V2 monoclonal antibodies show broad cross-reactivity41,48,68 (S. Zolla-Pazner, unpublished), suggesting that antibodies can target conserved elements in the V2 loop and that this region of Env might be pivotal for the induction of protective antibodies by successful vaccines. Since replacement of various aa in the V2 loop can alter the structure and function of the Env rendering HIV-1 less infectious or more sensitive to neutralization,33,48,69 the choice of Env and/or V2 sequences used in rational vaccine design should be carefully considered.
The authors thank SCHARP for assisting with the microarray analysis, James Swetnam for providing the biotinylated subtype B linear peptides, Kelly Soderberg for coordination of sample shipment and project management, Rob O'Connell for constructive reading of the manuscript, Robert Parks, Ryan Meyerhoff, and Krissey Lloyd for ELISA technical assistance, and Ellen Turk for performing the peptide microarray analysis. These studies were supported in part by an Interagency Agreement Y1-AI-2642-12 between U.S. Army Medical Research and Materiel Command (USAMRMC) and the National Institutes of Allergy and Infectious Diseases. In addition, this work was supported by a cooperative agreement (W81XWH-07-2-0067) between the Henry M. Jackson Foundation for the Advancement of Military Medicine and the U.S. Department of Defense. This research was funded, in part, by the U.S. National Institute of Allergy and Infectious Diseases. Peptide microarray work was funded by the Bill and Melinda Gates Foundation's Collaboration for AIDS Vaccine Discovery (OPP38744) and the NIH Vaccine Research Center. S.Z.P. and T.J.C. receive funding from NIH Grants HL59725 (S.Z.P.) and AI084119 (T.J.C.) and the Department of Veterans Affairs. Supported in part by grants from the Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery to Dr. Haynes.
MOPH-TAVEG Collaboration: MOPH: Dr. Supachai Rerks-Ngarm, Dr. Prayura Kunasol, Dr. Nakorn Premsri, Dr. Chawetsan Namwat, Prof. Prasert Thongcharoen Mahidol, Prof. Punnee Pitisuttithum, Dr. Jaranit Kaewkungwal. RTA: MG Sorachai Nitayaphan, MG (ret) Chirapa Eamsila. MHRP: Dr. Nelson L. Michael, Dr. Jerome H. Kim, Dr. Robert C O'Connell, Dr. Merlin L. Robb, Dr. Robert Paris, Charla Andrews, Dr. Jeffrey Currier.
AFRIMS: Dr. Viseth Ngauy, Dr. Mark S. de Souza, Dr. Nicos Karasavvas, Ms. Rapee Trichavaroj, Ms. Susan T. Mason, Ms. Bessara Nuntapinit, Ms. Nampueng Churikanont, Dr. Michael Benenson, Ms. Patricia Morgan.
Abstract presentations: AIDS Vaccine 2011: Global Collaboration and Coordination to Advance HIV Vaccine Research, September 12–15, 2011, Bangkok, Thailand.
Disclaimer: The views expressed in this article are those of the authors and do not reflect the official policy of the Department of the Army, Department of Defense, the Department of Veterans Affairs, or the U.S. Government.
No competing financial interests exist.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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CPRconstruction87 Feedback reviews, 100% positive
I am well established in my area, hard working, trust worthy and I will take the time with you to go through exactly what you need . I do an all-around construction trade service and specials in carpentry / joinery. No job is too big or small . I always pride myself in...
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By jules73_20 on 9th Jan, 2017
Upright Joinery30 Feedback reviews, 100% positive
hi,my name is frankie a joiner with 40yrs experience and i'm available for all types of joinery and building work.i usually work on my own these days but have other joiners and tradesmen available when required.i will travel to work anywhere for the smallest to the larger...
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By liz_eti on 16th Sep, 2017
S Mckenzie Joinery17 Feedback reviews, 100% positive
i have been a joiner for over 20 years undertaking all aspects of joinery to a high standard i undertake all aspects of joinery from interior to exterior 1st and second fix and work together with all clients to deliver required finish at a competitive price
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By patkin10 on 22nd Sep, 2017
MILLAR RENOVATIONS &PROPERTY MAINTENANCE53 Feedback reviews, 100% positive
My name is Derek & i run a renovations company, i am also a joiner to trade with many years of experience in all aspects of joinery & construction work. Within my company we also offer a single source solution to all projects which include fully qualified tradesmen.
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By mairij_74 on 4th May, 2017
SB Joinery12 Feedback reviews, 92% positive
I am a time served joiner but have also spent a lot of my career working between the workshop and in domestic and commercial buildings. I have more than 12 years experience in joinery. I cover all aspects of joinery and we pride ourself in being reliable and working to a...
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By mcl_create on 4th Aug, 2017
PM Joiners20 Feedback reviews, 100% positive
Qualified Joiner with 17years experience. Specialising in finishing works, internal doors and architraves. My company also supplies all trades while fitting kitchens, ensuring that my trusted GasSafe engineer, plaster and Electrician turn up at the required stages. Ensuring...
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By ally49 on 19th Apr, 2017
GMC Kitchens and Bedrooms11 Feedback reviews, 100% positive
kitchens and bedrooms supply and fit Remodels undertaken door replacement and worktops Integrated apps supply and refitted small joinery jobs undertaken
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By sheila_mcneill on 19th Sep, 2017
Kiwi and copper9 Feedback reviews, 100% positive
Hi I'm Josh owner of Kiwi and Copper. I am a new Zealand and moved to Northern Ireland in pursuit of a fiery red head. Hence kiwi and Copper, Now we're married and settled on the north coast. I am a bespoke furniture specialist creating custom pieces to client...
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By barbara75 on 28th Apr, 2017
HK Joinery and Property Maintenance6 Feedback reviews, 100% positive
We at Hk joinery pride ourselves in our attention to detail & customer satisfaction! Through the years we've built up a top class service of all trades! From design to completion Tell us What you'd like an we'll make it happen and more!
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By janet18 on 23rd Aug, 2016
Aldem5 Feedback reviews, 100% positive
We are Reliable, and Fair, We Like To See The Job First Before We Give a Price, We Can Get Building Control Certificates For All Loft Conversions, We Give Free Estimates. *******Very Keen Prices For Loft Conversions , or just get a Loft Ladder Fitted *******Great...
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Reference taken by MyBuilder on 5th Nov, 2009
argyll leisure pool systems4 Feedback reviews, 100% positive
What You dream about, new or old we build. 40 years of construction experience.Specialist approved installers, timber preservation and restoration experts. Swimming pool,hot tub sauna and steam room design and build contractor. Part time lecturer in construction at...
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By carlg1966 on 27th Apr, 2017
Handyman Me8 Feedback reviews, 100% positive
Mission statement: To provide a reliable and quality service, which the customer is always happy with. Open and honest advice is given to provide the best solution.
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By emxb on 1st Sep, 2013
Burke Construction4 Feedback reviews, 100% positive
We are a second-generation building firm of 3 brothers. Have been working in the construction industry for 15 years or more. Our father was a building contractor and we have just continued the family business so it really has been part of our life from a young age and we...
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By mullan on 18th Apr, 2017
D Mckenna1 Feedback review, 100% positive
Am at this game over 20 years . from houses, Hospatails,Schools,offices,Setting up for films ,shopfitting and ect
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By pdonnelly on 25th Jun, 2017
Bradley Contracts3 Feedback reviews, 100% positive
Here at Bradley Contracts we carry out work on residential, commercial and retail properties. Maintenance and new work from green field site to turn key completion. We have over 15 years experience in construction works since starting out in 2001. Every job is carried out to...
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By kevinq on 9th May, 2017
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subset: commoncrawl


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The ladies of Wellington Group love exploring Austin food, especially when it's gluten free. But as you probably know, finding good gluten free restaurants can be a hassle. To simplify things for all of you GF foodies, here's a list of our top 5 favorite gluten free spots in Austin. Happy eating!
- Gluten Free Buns from Hopdoddy Burger Bar
- Gluten Free Desserts from Better Bites Bakery
- Gluten Free Tartines from Blue Dahlia Bistro
- Gluten Free Pasta from Olive & June
- Gluten Free Pizza from Sagra Trattoria and Bar


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Ford dowel pin: 171ci. - 2.8L - V6: .940 x .570 - Photo ID 62934
Page Descritpion: Ford dowel pin: 171ci. - 2.8L - V6: .940 x .570 auto part photo. Engine auto part photos, Engine Camshaft Parts auto part photos, Engine Camshaft Dowel Pin auto part photos. Ford auto part photos. Image gallery of car part pictures
Page Keywords/Tags: Engine Camshaft Dowel Pin Images, Ford Car Photos, Engine Camshaft Dowel Pin Auto Parts Images, Ford Images, Engine Camshaft Dowel Pin Car Photos, Ford Parts Pictures
|©Copyright 1998-2017
All World, Inc.
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|NOTICE: All World, Inc. is a venue for companies wishing to buy or sell their surplus auto parts inventories. Names, brands and products named on these pages have registered trademarks belonging to their respective owners. Representation of inventories on these pages is the responsibility of the listing companies.


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Cold stays...rain returns!
We do have some sleety mix in our southern counties this AM, but most of us will stay dry as another cold front plows through. Winds will pick up later AM from the north and keep temps in the 30s today.
A cold DRY night is expected as temps fall well into the 20s...even some teens north. After a dry start Sat (even some breaks in the clouds), clouds thicken and we could see dome sleet/rain mix develop PM esp south of DFW.
That would spread in during the evening as a rain OR freezing rain mix as temps will be close to freezing.
That will continue Sunday AM as a cold rain...then end midday or afternoon as we get back to near 40.
Next week is chilly, but not AS cold, with lots of clouds Mon-Tue and a chance for some light rain Tuesday night.


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Tumi Lawrence aka SliZz Da WhiZz, is a professional broadcaster with an endearing airwaves attitude and began his radio career in 2004. Such expertise of his has been garnered over the years with working experience from different radio stations he has worked with covering different belts and styles of radio programming. Over the years, he has hosted sponsored syndicated radio shows aired on a number of radio stations across the nation. He recently won the award for the Radio Presenter Of The Year at the EYA Award. Before joining Star FM Lagos, he…Read More
Category: SliZz Da WhiZz
Sliz Da Wiz
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Posted January 21, 2014 in -Ended
Holler If Ya Hear Me
Featuring rapper Tupac Shakur's music and lyrics, Holler If Ya Hear Me offers a powerful story focusing on the lives of two childhood friends. Acclaimed director and Tony nominee Kenny Leon (2010 Fences) will bring this new musical to life, along with choreographer Wayne Cilento. Leon, who says the project is the "greatest artistic challenge" of his life, has called Tupac Shakur's music a "mountain of beauty." Holler If Ya Hear Me looks to be one of the most innovative musicals that Broadway has seen in a long time.


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by Keith Lokkesmoe
Special to Sun Thisweek
Dakota County Tribune
Sex trafficking is none other than the business of selling and buying women, girls and boys to be sexually used and abused by men. It is not just a problem occurring overseas or in inner cities. It is happening in every city and every suburb in the Twin Cities and many towns across Minnesota. This menace has the potential to rob our sons, daughters, nieces, nephews, friends, and grandchildren of not only their innocence, but their future.
When people think of sex trafficking, they usually think of the victims — the survivors. But sex trafficking would not occur if men were not purchasing sex. In order to stop the exploitation of trafficking, we must reduce the demand. What is behind this demand? It starts with what people see. Scantily clad women are used to sell products on television. The Sports Illustrated swimsuit edition is sold to men by women selling their air-brushed bodies as symbols of value. Sitcoms and movies joke about bachelor parties with "hookers" as being normal. College football team members sexually use and abuse a woman and believe their actions are justified.
One step deeper in driving demand lies the Commercial Sex Industry. This conglomeration of pornography sites, adult entertainment businesses, escort services, and strip clubs is a multi-billion-dollar industry. Men and boys who've seen scantily clad women in Sports Illustrated or nudity in a mainstream movie now want something more. So, they begin searching their laptops and smartphones for porn sites that will give them the graphic stuff — movies, pictures, chat sites — which inflame their desire and twist their minds into thinking that women like this stuff.
Finally, for many, simply watching is not enough anymore. They need an actual encounter with a woman. So, they venture into strip clubs, sex clubs, and ultimately search for "escort" websites. Hundreds of ads appear each week on these sites, and each new ad has men calling within three minutes of posting. The vast majority of women being prostituted today have been sexually abused as a child; 99.9% of them are completely controlled and abused by their pimps. They are not free women working in dignity. They are daily assaulted by men thinking that they have every right to use and abuse these women as they see fit.
If we want to stop sex trafficking, we need to reduce the frequency at which men pick up the phone or click on their computer to order sex. What to do? Here are six areas we can focus on:
1. Expose pornography as a public health issue. In April 2016, Utah became the first state to declare pornography as a public health issue. All states should follow suit. Dr. Mary Anne Layden, director of the Sexual Trauma and Psychopathology Program Center at the University of Pennsylvania, has researched the violent nature of today's pornography and concludes that the use of pornography leads to more acceptance of violence against women, creating more sexually violent fantasies to get aroused, engaging in more sexual harassment behaviors, and a greater likelihood to use physical coercion to have sex.
2. Expose strip clubs as harbors of abuse and forerunners of prostitution. We have many first-hand reports of the daily abuse occurring in these establishments.
3. Protect young people from internet and mall traffickers.
Predators have developed sophisticated strategies for convincing young girls to meet with them, such as posing as a teenage boy. They also obtain nude pictures from teens and use these to blackmail and exploit. Sex traffickers also routinely work at shopping malls, coffee shops — anywhere that teenage girls frequent. We need to educate teens on these methods.
4. Change the laws to make hard core pornography illegal. There are national obscenity laws against pornography. There have been some challenges to enforcement of obscenity laws based on First Amendment concerns. However, prosecutors today, in light of the dire harm being caused by pornography and its link to trafficking, would do well to take up this issue with renewed vigor.
5. Increase the penalties for persons who buy or sell people. Minnesota is a national leader in terms of passing and funding Safe Harbor legislation, which has been expanded upon to now provide services for women or men being prostituted until age 24. We need to ensure that new legislation increases the penalties for both buyers and traffickers to felony level status.
6. Confront the mindset that women are sex objects and men are helpless to control themselves. Rape culture asks the woman first what she did to encourage the man. It assumes that the women's answer to sexual advances is yes — unless there is clear and convincing evidence otherwise. It absolves men of responsibility and casts them as unable to control their desires. We can do better than this.
This is a dark and complex problem. But it is not unsolvable. Only if the American public remains ignorant, will the traffickers stay in business. Once the majority of Americans are made aware of this menace, that it stalks their very own daughters and sons and brothers and sisters and grandchildren, we can all rise up and begin to stop this plague in our midst. Please join us in this battle.
Trafficking Justice will have its Freedom Weekend March 17-19, which will provide specific information on this problem and what people can do about it. The weekend is a collection of seven different events at multiple locations in the Twin Cities. There will be international, national, and local speakers who are working at the front lines of the battle against sex trafficking today. All events are free.
Go to www.traffickingjustice.com/Freedom-Weekend.html for all of the details.
Keith Lokkesmoe, of Lakeville, is the executive director of Trafficking Justice — a group in the Minneapolis area comprised of churches, faith-based organizations, and individuals who have come together to fight for freedom from "modern-day slavery" known as human trafficking. Columns reflect the opinion of the author.


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Rattan Diffuser Reeds - 100 Bulk pack
$39.95 / 100 packQty:
Angel Aromatics have all your reed diffuser supplies. Our diffuser reeds are made from rattan and is available in a bulk pack of 100.
Rattan diffuser reeds diffuse the fragrance by allowing it to draw the scent upwards and out into the air.
Also available in smaller size and in a 10 pack of reeds - see more products below.
Click here to view more reed diffuser supplies (sets and refills).Additional Information
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Angel Stock Location: 993
Gorgeous home fragrances, handmade in Australia. Angel Aromatics are an Australian manufacturer based in Sydney. Decorative scented homewares to fragrant candles, oils and sprays. All home fragrance products are Made in Australia using the highest quality fragrances and essential oils which are paraben free, vegan friendly and not tested on animals. Home fragrance ranges include: balsa wood decorative scented flowers, timber curls, diffusers, oils, sprays, candles, melts. Try our famous Annan scent. Custom Manufacturing and wholesale bulk supply available.


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After a break of a few years 16 of us in a Painting Group returned again this year to this lovely hotel, right on the beach which we were delighted to find has continued to stay abreast of the times. The refurbishments carried out over...More
Getting you more information on this room
Getting you more information on this room
"This hotel has a variety of different rooms from single use, to penthouse suite. Sea or Mountain views."
"With a sea view room you can listen to the waves lapping on the short and watch the world go by"
"Sea view rooms have excellent views, but the other rooms have good views of the mountains."


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Mint Unused World War II US Army Field Dressing, two packages, shrink wrapped for protection. Not for consumption or use. Item is for display only. Made by Handy Pad Supply Co., of Worcester, Mass. These packets are the Large Carlisle First-Aid Dressing, 4" by 2 1/4". See photos.
This is the last of a massive 12k piece antique pharmacy collection. Some of the best saved for last. It will take us a while but we will be actively adding items. So in addition to the 150+ pharmacy and drugstore items available for sale right now we will be adding more. All are ON SALE, tins, glass bottles, apothecary jars, unopened bottles in original boxes. All bottles are clean. NO damage, NO cracks, NO chips.
To purchase with MC or VISA please call Drury House Antiques 802-824-4395 EST or use PAYPAL
Need help with your order, Call us 802-8244395 EST 8 AM to 8 PM
Sale Final Days


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Hearing aids in Cherry Hill, NJ
We found 40 hearing clinics with 210 reviews within 21.3 miles of Cherry Hill, NJ. Show clinics near me.
Hearing professionals near you
Total Hearing Care - Cherry Hill (1.7 miles)
1050 N Kings Hwy Ste 101 Cherry Hill, NJ 08034
Elizabeth Cook, MA, FAAA
Elizabeth Cook is a truly dedicated professional audiologist in NJ who brings over 30 years of (read more)
H A Mckee & Associates (2 miles)
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HearMD - Haddonfield (2.4 miles)
130 N Haddon Ave Haddonfield, NJ 08033
Dorothy DeFeo, M.A., CCC-A
Dorothy earned her Masters of Arts degree and her CCC-A - Certificate of Clinical Competence (read more)
Associates In Hearing HealthCare - Barrington (4.2 miles)
121 Clements Bridge Rd Barrington, NJ 08007
Elizabeth Kennedy, Ph.D., CCC-A
Dr. Elizabeth Kennedy is the principal owner of Associates in Hearing Healthcare, a private practice (read more)
Audiology Partners, LLC (5.2 miles)
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Audiology Partners LLC (5.3 miles)
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HearMD - Voorhees (6.1 miles)
200 Bowman Dr D-285 Voorhees, NJ 08043
Caryn Bross, MS, CCC-A
Caryn Bross received a Master of Science degree in Audiology from Teachers College, Columbia (read more)
REM Audiology Associates, P.C. - Voorhees (6.4 miles)
1000 White Horse Rd Ste 510 Voorhees, NJ 08043
Elizabeth Patterson, AuD, CCC-A, FAAA
With over 35 years of experience in the area of audiology, Elizabeth Protti-Patterson has developed (read more)
Becker Hearing Center (6.7 miles)
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Jefferson Hearing and Balance Center (6.8 miles)
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Main Line Audiology Consultants - Ce… (7.5 miles)
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Advanced Hearing Systems Inc - Phila… (7.6 miles)
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Associates in Hearing Healthcare - Marlton (7.8 miles)
406 Lippincott Dr Ste G Marlton, NJ 08053
Elizabeth Kennedy, PhD, CCC-A, FAAA
Elizabeth has worked with individuals with hearing loss for over 25 years. She has worked in a (read more)
Temple Head & Neck Institute (8.1 miles)
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REM Audiology Associates P.C. - Marlton (8.5 miles)
775 Route 70 East Suite F 120B Marlton, NJ 08053
Maegan Mapes, Au.D., CCC-A
Licensed Doctor of Audiology Maegan Mapes earned her bachelor's degree at the University of (read more)
REM Audiology Associates P.C. - Phi… (8.6 miles)
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HearMD - Mount Laurel (8.7 miles)
204 Ark Rd Ste 102 Mount Laurel, NJ 08054
Laura Berman, M.S., CCC-A
Laura is a licensed and highly qualified Audiologist for this location. She earned her Masters (read more)
HearMD - Woodbury (8.8 miles)
620 N Broad St Ste 1 Woodbury, NJ 08096
Laurie Denisi, BA, HIS
Laurie Denisi received her Bachelor of Science Degree from St. Joseph's University. She is a (read more)
Temple Head & Neck Institute (9.7 miles)
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Aria Health Torresdale Campus (9.9 miles)
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Melissa Schwartz, DO, LLC/Montgomery… (10.3 miles)
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HearMD - Medford (10.8 miles)
103 Old Marlton Pike Ste 219 Medford, NJ 08055
Linda Goldstein, AuD, CCC-A
Linda earned her Audiology degree from Salus University and her M.A. degree from Temple University. (read more)
HearMD - Willingboro (11.1 miles)
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Listen 2 Life Hearing Center - Phila… (11.4 miles)
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PA Ear Institute (11.7 miles)
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Main Line Audiology Consultants - Je… (12.4 miles)
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Hearing Hut (13 miles)
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Main Line Audiology Consultants - Narberth (13.3 miles)
916 Montgomery Ave Narberth, PA 19072
Kathy Landau-Goodman, Au.D.
Dr. Kathy Landau Goodman earned her Doctorate degree in Audiology from Central Michigan University (read more)
Beaver Hearing Aid Center (13.3 miles)
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Listen 2 Life Hearing Center - Phila… (13.8 miles)
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Ask Physicians Hearing Centers (14.5 miles)
301 W Chester Pike Ste 101 Havertown, PA 19083
Cindy Connor, AuD, CCC-A, Director of Audiology
Licensed doctor of audiology Margaret (Cindy) Connor is the Director of Audiology at Head (read more)
HearMD - Sewell (14.8 miles)
239 Hurffville Crosskeys Rd Ste 265 Sewell, NJ 08080
Drexel Hill Hearing Aid Center (15.2 miles)
4801 State Rd Drexel Hill, PA 19026
David Epstein, HIS
Mr. David Epstein is a state-licensed hearing aid dispenser offering quality hearing aids powered by (read more)
Listen 2 Life Hearing Center - Willo… (15.9 miles)
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Temple Head & Neck Institute (16.6 miles)
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Advanced Hearing Systems Inc (18.1 miles)
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Audiology and Hearing Aid Center (18.5 miles)
65 W Street Rd Suite B104 Warminster, PA 18974
Holli Lish, Au.D.
Holli has been working in the field of audiology for over 20 years, employed in both private (read more)
Premier ENT Associates Bucks Mercer … (19.2 miles)
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Boricua College forestry department ranking
Think about what career is best suited for you, and where you want to study. Request more information now.
Think about what career is best suited for you, and where you want to study. Request more information now.
Think about what career is best suited for you, and where you want to study. Request more information now.
Get the facts about Boricua College forestry department ranking, and registering for elective seminars. Which college degrees give you the best chances of finding employment after graduation? Nursing is one of the fastest-growing job areas, and for good reason. As the population ages, medical care will continue to expand rapidly. Concentrating on Boricua College forestry department ranking, for example, you may gain an advantage in job recruiting on-campus. Check with the environment department for course registration deadlines, and forestry program requirements. Please use the form above to request admissions info for Boricua College.
Agricultural biotechnology, and crop science in general, includes plant cross-breeding techniques, trait selection, and genetic engineering in some cases. Biotech crops can make farming more profitable by increasing crop quality and may in some cases increasing crop yields. Where biotech crops are grown in proximity to related plants, the potential for the two plant species to exchange traits via pollen must be evaluated. In fact, according to the USDA National Agricultural Statistics Service (NASS), herbicide-tolerant crops are particularly compatible with reduced-tillage commercial agriculture systems that help preserve topsoil from erosion.
USDA researchers seek to solve major agricultural problems and to better understand the basic plant biology relating to crop science. Researchers may use biotechnology processes to discover things that may not be possible by more conventional means. This includes introducing new or improved traits in plants, animals, and microorganisms and creating new biotechnology-based products such as more effective diagnostic tests, improved farm animal vaccines, and better antibiotics. The USDA Economic Research Service (ERS) conducts research on the economic aspects of the use of genetically engineered plants, including the rate of adoption of biotechnology by farmers. The regulation of genetic engineering aims to assess and manage the risks of developing genetically modified organisms (GMO), including commercial food crops and fish. Beyond major requirements within the environment department, you may consider Siena Heights University restaurants as a complementary elective to your primary course of study.
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My little trip!
So, this August I took a little trip across western Europe. After all my travels abroad I thought it was about time I went to visit some of the awesome friends I have met at Fashion Weeks and have a luxury inter railing trip...
I think my trip was more luxury than standard because:
a) I didn't stay in one fucking hostel. Do I look like I stay in hostels?
b) I was visiting Fashion Friends Darling
c) I did a lot of extra work and saved a lot of money (all gone now!)
So now that settled... on with the tour. (If I come across as smug its because I am)
Waking up late on the morning I caught the Eurostar made this morning totally awesome... not.
I arrived in Paris and met Jean Luc (@Systeme_D) – after a hearty breakfast we hit Versaille, my trademark beehive intact and wandered through the grounds and palace wondering why the visitors weren't bowing to me as with my hair and stature I am clearly Marie Antoinnette reborn. No one even asked if I had any cake.
The next day (after another hearty breakfast to soak up the previous evenings wine) I hit Le Marais. Paris's best vintage shopping quarter. I emerged significantly better off with bags full of gorgeous clothes (including a shin length A-Line leather skirt for 20euros) – this is when I wandered star struck into Adidas and found the Jeremy Scott collection on sale. 125 Euros you say? Done!
We headed to HD Diner for a highly alcoholic milkshake then to the Helmut Newton exhibition. This photographer was incredibly before his time and so many of his shots it is hard to believe were taken before the digital era. Really enlightening exhibition (also depressing.... my work will never be that good). If you are in Paris before the exhibition ends it is well worth a visit.
As we were leaving the discovery was made that the mens toilet matched my jacket perfectly. An impromptu photoshoot ensued.
My last day in Paris was a rather morbidly cultural trip to the Père Lachaise Cemetery where such 'celebs' as Oscar Wilde are buried. His tomb is surrounded by graffitied Perspex and the tomb itself is hideous. Shame. Jim Morrisons grave is littered with whiskey bottles and the gravestones around it are graffitited with messages to the dead legend. This seemed really, really wrong to me.
People could ignore the resting places of other humans to scrawl a message to a dead guy without a thought to the families of the surrounding people.
My train to Madrid was pretty un-interesting apart from the sullen Japanese girl who refused a glass of wine like I was a pedophile and tried to eat an entire can of peaches out of a plastic cup with out a spoon....
Arriving in Madrid I discovered that my hotel was well above its 2* standard. Clean and everything was brand new. I was expecting a little bit of single room scum but there weren't even pubes on the sheets! (I kid... I do genuinely have standards.)
After visiting the gorgeous Royal Palace in Madrid (apparently bigger than Versaille) I made my way to Retiro Park – a huge sprawling park with an amazing parthanon inspired creation over looking a lake and a palace built on the basis of Crystal Palace – yes I went all the way to Spain to hang out in Crystal Palace. The lakes were home to rowing boats and some snappy little terrapins that kids were feeding with hotdogs. Despite being a vegetarian I was considering buying a hot dog to join in this fun. I then bumped into two awesome German girls that were on the same train as me from Paris and continued to ruin there trip with my blatant English-ness plying them with rum and partying the night away in empty clubs. We know how to do it.
Then on to Lisbon!
(Where no one wakes up before midday – remember that when you are expecting to meet up with people. 11am means 2pm.)
(Where no one wakes up before midday – remember that when you are expecting to meet up with people. 11am means 2pm.)
I was staying with the wonderful Fiona Bunnett – a charming Gal originally from Durham or Newcastle.... somewhere northern who moved to Portugal where she is a model, journalist, celebrity etc. In addition to this she is fucking awesome.
My first day I got incredibly lost in Lisbon for hours, eventually ending up at the Castle Jorge where the views are really quite spectacular despite the tourists and dudes pedalling strings of beads.
Next I headed to the Guava Showroom (post to come) where I also met the wonderful Valentim Quaresma and had a look around his studio and showroom.
The evening consisted of a classy sunset drink at Silk Bar on the penthouse balcony (Oh I do say!) with Fiona and Tiago (Miranda – head of ModaLisboa International Press i.e. god, when you are at modalisboa and a tourist). Next was the Martini party who was holding a pagent for Miss Martini! This was hosted by a very famous Portuguese comedian (whose name escapes me...) didn't understand any of his jokes but he set the lovely ladies tasks. They were excruciating 'I hope your children/parents don't see this on TV' tasks including sexily pouring and serving a glass of martini (bottle neck stroking...) and tempting a man tom buy them a drink – grinding, hair swishing and lip-licking were a must for most of these tasks.
After helping to drink the bar dry we headed home for Pizza and a girly chat on the sofa!
The next day I spent another fruitless day sunning myself and not getting a tan (seriously.) before heading out for dinner and drinks with Vitor, Marcio, Diogo and Ines at a lovely traditional restaurant called 'Buenes Aires'. This was all washed down with around a gallon of wine and sangria.
Fiona joined us and we headed to Lux.... I have distant half memories of being very drunk in this club during ModaLisboa.
Myself and Vitor let lose here on beer and classy attendees danced up a storm... then at 5.30am my body clock utterly failed and literally I could not wait to get home and go to bed!
My next evening was an impromptu dinner party at Mia's house. Mia is one half of the design duo Os Burgueses who's last collection was a sophisticated mix of urban metal and military chic.
We sucked a wine box dry and then tormented the poor frequenters of Grindr with this photo of Vitor. Still want some milk on your face?
My final evening was a trip to the beach with the lovely Ines and Diogo before heading to another dinner get together, drinks in a square and then a drum and bass club 'Ritz'.
In short, a blur of drinks, parties and lounging in the sun desperately trying to make my 'porcelain' skin slightly more terracotta.
Onwards to the Algarve!
These final days were fab!
First night there was a famous beach party, before heading to Wonderful Beach club!
My final day in the Algarve we went to Tavira island and spent the day on the beach and Fiona and Helio (Barbie and Ken!) had a pretty impressive sea food dish.
Then of course, we partied until dawn again.....
Goodbye Europe, I'll be back soon...
Thank you so much to Jean Luc and Fiona for letting me stay and to everyone else for coming out to see me, putting up with my complete lack of language skills and for generally helping me have an amazing holiday.
Much Love
Jemima Daisy x
Onwards to the Algarve!
These final days were fab!
First night there was a famous beach party, before heading to Wonderful Beach club!
My final day in the Algarve we went to Tavira island and spent the day on the beach and Fiona and Helio (Barbie and Ken!) had a pretty impressive sea food dish.
Then of course, we partied until dawn again.....
Goodbye Europe, I'll be back soon...
Thank you so much to Jean Luc and Fiona for letting me stay and to everyone else for coming out to see me, putting up with my complete lack of language skills and for generally helping me have an amazing holiday.
Much Love
Jemima Daisy x


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Firehawks Carry Sharp to Long-Awaited Victory Scott Sharp took the checkered flag today at Kentucky Speedway in the AMBER Alert Portal Indy 300 for his first victory since 2003. Sharp, driving for Fernandez Racing, started in seventh position...
Firehawks Carry Sharp to Long-Awaited Victory
Scott Sharp took the checkered flag today at Kentucky Speedway in the AMBER Alert Portal Indy 300 for his first victory since 2003. Sharp, driving for Fernandez Racing, started in seventh position and took the lead late, beating Vitor Meira across the finish line by just 0.0779 of a second.
Sharp won on a Firestone Firehawk tire specification used twice already this season--at Texas Motor Speedway and Kansas Speedway. But the low wear rate at Kentucky Speedway, along with incredibly hot temperatures, turned out to be a little tougher on the tires and some teams experienced problems during the race.
Despite the harsh conditions, the tires held up well throughout and the race winner had no complaints about their performance.
"I gotta tell you, I think the biggest constant in the IRL is Firestone and their tremendous support," said Sharp. "I've driven other tires, and I can tell you that Firestone tires are consistently flawless. From start to finish today, from lap one to lap 50, they were tremendous."
Quote from Joe Barbieri, Manager, Firestone Racing: "This one turned out to be quite a challenge. Having run this same spec at two different tracks this year we weren't expecting any problems, but conditions were pretty harsh, in terms of tires, all weekend long. Most teams were able to make adjustments and, in the end, the tires held up and it turned out to be a fine show for the fans here at Kentucky. It was great to see Scott Sharp and Fernandez Racing back in the winner's circle."
Firestone Performance Award winner: Vitor Meira of Rahal Letterman Racing led lap 115 today to claim the Firestone Performance Award, which commemorates the fact Firestone has competed in every IRL IndyCar Series event since the league's 1996 inception. Today's race was the 115th in league history. Meira will be presented a $10,000 check during pre-race festivities for the Honda Indy 225 at Pikes Peak International Raceway on Aug. 21.
Race temperatures ranged from 109 to 122 degrees track, with a constant ambient of 86 degrees (F).
Race temperatures ranged from 43 to 50 degrees track, with a constant ambient of 30 degrees (C).


text: Zedge Company Profile (NASDAQ:ZDGE) Company Profile Analyst Ratings Earnings History Earnings Estimates Dividend History Insider Trade Headlines Social Media Chart About Zedge (NASDAQ:ZDGE) Zedge, Inc. is a provider of content distribution platforms, centered on self-expression, enabling both creators looking to promote their content and consumers utilizing such content to express their identity, feelings, tastes and interests. The Company's platform enables consumers to personalize their mobile devices with free, ringtones, wallpapers, home screen application icons and notification sounds. The users primarily access the Company's content through its smartphone application, called Zedge. The Company's smartphone application is available in the Google Play and iTunes application stores. The Company has developed a distributed platform that consists of both open source and proprietary technologies centered on content management and discovery, Web and application development, data mining and analytics, deep learning, mobile content/device compatibility, advertising and reporting. Industry, Sector and Symbol: Sector: Technology Industry: Mobile Application Software Sub-Industry: N/A Symbol: NASDAQ:ZDGE CUSIP: N/A Web: N/A Capitalization:Market Cap: $18.43 millionOutstanding Shares: 9,647,000Average Prices:50 Day Moving Avg: $1.85200 Day Moving Avg: $2.5452 Week Range: $1.65 - $4.36 P/E:Trailing P/E Ratio: N/AP/E Growth: 0.00Sales & Book Value:Annual Revenue: $9.94 millionPrice / Sales: 1.85Book Value: $1.09 per sharePrice / Book: 1.75 Profitability:EBIDTA: ($277,000.00)Net Margins: -9.45%Return on Equity: -9.04%Return on Assets: -7.62%Misc:Average Volume: 22,852 shs.Short Ratio: 1.29 Frequently Asked Questions for Zedge (NASDAQ:ZDGE) What is Zedge's stock symbol? Zedge trades on the NASDAQ under the ticker symbol "ZDGE." Who are some of Zedge's key competitors? Some companies that are related to Zedge include Windar Photonics PLC (WPHO), Sanderson Group plc (SND), Cerillion PLC (CER), MobileSmith (MOST), Scisys Plc (SSY), Trakm8 Holdings PLC (TRAK), Inuvo (INUV), Intercede Group plc (IGP), Sopheon Plc (SPE), Instem PLC (INS), Global Invacom Group Ltd (GINV), Artilium plc (ARTA), Cyanconnode Holdings PLC (CYAN), ENTREC Corp (ENT), EG Solutions plc (EGS), Audioboom Group PLC (BOOM), Redstoneconnect PLC (REDS) and Universe Group plc (UNG). How do I buy Zedge stock? Shares of Zedge can be purchased through any online brokerage account. Popular online brokerages include Vanguard Brokerage Services, TD Ameritrade, E*TRADE, Scottrade, Fidelity and Charles Schwab. What is Zedge's stock price today? One share of Zedge stock can currently be purchased for approximately $1.91. MarketBeat Community Rating for Zedge (NASDAQ ZDGE)Community Ranking: 2.5 out of 5 ( )Outperform Votes: 28 (Vote Outperform)Underperform Votes: 29 (Vote Underperform)Total Votes: 57MarketBeat's community ratings are surveys of what our community members think about Zedge and other stocks. Vote "Outperform" if you believe the stock will outperform the S&P 500 over the long term. Vote "Underperform" if you believe the stock will underperform the S&P 500 over the long term. You may vote once every thirty days. Analyst Ratings Consensus Ratings for Zedge (NASDAQ:ZDGE) (How are Consensus Ratings Calculated?) MarketBeat calculates consensus ratings using the most recent rating from each brokerage that has rated a stock within the last twelve months. Since brokers often use different ratings systems, each rating is normalized to a standardized rating score of 1 (sell), 2 (hold), 3 (buy) or 4 (strong buy). Consensus ratings scores are calculated using the mean average of the number of normalized sell, hold, buy and strong buy ratings. Each stock's consensus rating is derived from its calculated consensus ratings score (0-1.5 = Sell, 1.5-2.5 = Hold, 2.5-3.5 = Buy, >3.5 = Strong Buy). MarketBeat's consensus price targets are a mean average of the most recent available price targets set by each analyst that has set a price target for the stock in the last twelve months.MarketBeat will no longer include ratings and price target data in its consensus calculation if a broker drops coverage and MarketBeat has received a report that coverage was dropped. Ratings from certain research firms that issue ratings using purely quantitative methods (such as Zacks, Vetr and ValuEngine) are not included in consensus calculations. MarketBeat's consensus ratings and consensus price targets may differ from those calculated by other firms due to differences in methodology and available data.Ratings Breakdown: No ratings tracked in the last 12 months.Consensus Rating:N/A (Score: NaN)Consensus Price Target: N/A Analysts' Ratings History for Zedge (NASDAQ:ZDGE) Show: Only the Most Recent Rating From Each Brokerage All Ratings For This Stock No equities research coverage for this company has been tracked by MarketBeat.com Earnings Earnings History for Zedge (NASDAQ:ZDGE)Earnings History by Quarter for Zedge (NASDAQ ZDGE)DateQuarterConsensus EstimateActual EPSRevenue EstimateActual RevenueRelease LinkConference CallDetails6/13/2017Q3 2017($0.02)$2.53 millionViewN/A9/29/2016Q416($0.08)$2.50 millionViewListen(Data available from 1/1/2011 forward) Estimates Earnings Estimates for Zedge (NASDAQ:ZDGE)No earnings estimates for this company have been tracked by MarketBeat.com Dividends Dividend History for Zedge (NASDAQ:ZDGE)No dividend announcements for this company have been tracked by MarketBeat.com Insider Trades Insider Trading and Institutional Ownership History for Zedge (NASDAQ:ZDGE)Insider Trades by Quarter for Zedge (NASDAQ:ZDGE)Transaction DateInsider NameTitleBuy/SellNumber of SharesAverage Share PriceTotal TransactionDetails3/21/2017Stephen M. GreenbergDirectorSell5,783$3.26$18,852.58 3/17/2017Howard S JonasDirectorBuy6,432$3.36$21,611.52 3/14/2017Howard S JonasDirectorBuy722$3.40$2,454.80 3/13/2017Howard S JonasDirectorBuy3,766$3.39$12,766.74 3/9/2017Howard S JonasDirectorBuy5,892$3.37$19,856.04 3/6/2017Howard S JonasDirectorBuy3,771$3.39$12,783.69 2/27/2017Howard S JonasDirectorBuy1,990$3.36$6,686.40 2/15/2017Howard S JonasDirectorBuy1,700$3.40$5,780.00 2/7/2017Howard S JonasDirectorBuy1,761$3.39$5,969.79 2/6/2017Howard S JonasDirectorBuy10,607$3.37$35,745.59 2/2/2017Howard S JonasDirectorBuy18,686$3.32$62,037.52 1/30/2017Howard S JonasDirectorBuy23,037$3.30$76,022.10 1/26/2017Howard S JonasDirectorBuy26,733$3.29$87,951.57 1/23/2017Howard S JonasDirectorBuy15,849$3.25$51,509.25 1/19/2017Howard S JonasDirectorBuy14,206$3.21$45,601.26 1/17/2017Howard S JonasDirectorBuy15,818$3.22$50,933.96 1/12/2017Howard S JonasDirectorBuy19,702$3.18$62,652.36 1/9/2017Howard S JonasDirectorBuy12,706$3.21$40,786.26 1/5/2017Howard S JonasDirectorBuy12,582$3.20$40,262.40 1/3/2017Howard S JonasDirectorBuy16,076$3.07$49,353.32 12/28/2016Howard S JonasDirectorBuy14,391$3.04$43,748.64 12/27/2016Howard S JonasDirectorBuy5,891$3.05$17,967.55 12/23/2016Howard S JonasDirectorBuy3,800$3.06$11,628.00 12/22/2016Howard S JonasDirectorBuy14,019$3.08$43,178.52 12/19/2016Howard S JonasDirectorBuy11,457$3.19$36,547.83 12/12/2016Howard S JonasDirectorBuy100$3.25$325.00 12/9/2016Howard S JonasDirectorBuy2,300$3.20$7,360.00 11/15/2016Howard S JonasDirectorBuy2,922$3.07$8,970.54 11/14/2016Howard S JonasDirectorBuy9,446$3.07$28,999.22 11/10/2016Howard S JonasDirectorBuy15,967$2.95$47,102.65 11/7/2016Howard S JonasDirectorBuy5,100$3.00$15,300.00 11/3/2016Howard S JonasDirectorBuy10,923$3.00$32,769.00 10/19/2016Howard S. JonasDirectorBuy17,457$3.16$55,164.12 10/18/2016Howard S JonasDirectorBuy16,998$3.00$50,994.00 10/17/2016Howard S JonasDirectorBuy15,823$3.00$47,469.00 10/14/2016Howard S. JonasDirectorBuy25,871$2.75$71,145.25 (Data available from 1/1/2013 forward) Headlines Headline Trends for Zedge (NASDAQ:ZDGE) Latest Headlines for Zedge (NASDAQ:ZDGE) Source: All Sources Trusted Sources MarketBeat.com Bloomberg MarketWatch Motley Fool NASDAQ Reuters Seeking Alpha The Street The Wall Street Journal Yahoo Finance DateHeadlineETFs with exposure to Zedge, Inc. : August 24, 2017finance.yahoo.com - August 25 at 1:00 PMZedge, Inc.: Gathering momentum, can it sustain its performance?finance.yahoo.com - August 15 at 4:13 PMETFs with exposure to Zedge, Inc. : July 11, 2017finance.yahoo.com - July 11 at 3:21 PMZedge, Inc. :ZDGE-US: Earnings Analysis: Q3, 2017 By the Numbers : June 16, 2017finance.yahoo.com - June 17 at 8:02 AMETFs with exposure to Zedge, Inc. : June 14, 2017finance.yahoo.com - June 14 at 4:30 PMZedge Inc (ZDGE) Posts Quarterly Earnings Resultswww.americanbankingnews.com - June 14 at 9:36 AMZedge Announces Third Quarter Fiscal 2017 Resultsfinance.yahoo.com - June 14 at 2:27 AMZedge to Report Third Quarter Fiscal 2017 Resultsfinance.yahoo.com - May 31 at 11:27 AMTop Brands Utilizing Zedge Collections to Gain Access to the Smartphone Home Screenfinance.yahoo.com - April 27 at 11:25 AMZedge to Present at the Joseph Gunnar Pioneers 2017 Conferencefinance.yahoo.com - April 26 at 8:07 AMZedge Launches Premium Ringtone App on iTunes Featuring Marimba Remixes of Chart Topping Music from Drake, Ed Sheeran and Other Leading Artistsfinance.yahoo.com - April 25 at 8:23 AMZedge Introduces a Deeper Personalization Experience to iPhone Usersfinance.yahoo.com - April 7 at 1:40 PMZEDGE, INC. Financialsfinance.yahoo.com - March 24 at 2:50 AMInsider Selling: Zedge Inc (ZDGE) Director Sells 5,783 Shares of Stockwww.americanbankingnews.com - March 23 at 4:29 PMInsider Buying: Zedge Inc (ZDGE) Director Buys 6,432 Shares of Stockwww.americanbankingnews.com - March 20 at 4:55 PMZEDGE, INC. Files SEC form 10-Q, Quarterly Reportbiz.yahoo.com - March 17 at 3:35 PMZedge, Inc. (ZDGE)seekingalpha.com - March 17 at 5:33 AMZedge Announces Second Quarter Fiscal 2017 Resultsfinance.yahoo.com - March 17 at 5:33 AMZEDGE, INC. Files SEC form 8-K, Results of Operations and Financial Condition, Change in Directors or Principal Officbiz.yahoo.com - March 17 at 5:33 AMZedge Inc (ZDGE) Director Howard S. Jonas Buys 5,892 Shareswww.americanbankingnews.com - March 10 at 1:00 AMZedge to Report Second Quarter Fiscal 2017 Resultsfinance.yahoo.com - March 2 at 6:17 AMZEDGE, INC. Files SEC form 8-K, Submission of Matters to a Vote of Security Holdersbiz.yahoo.com - January 20 at 6:20 PM Social Chart Zedge (ZDGE) Chart for Monday, September, 25, 2017 This page was last updated on 9/25/2017 by MarketBeat.com Staff


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I am new to Teradata. Can some one help me to convert Sqlserver functions in to Teradata Function. I really appreciate for your help.
Hi Sasha,
Whaw!!!! All I can think of now is providing the links for you to see and code. You can download the docs.
You know the functionalities better than anyone else. I always suggest people that if we migrate from one DB to another DB, it is better not to think much of code by code unless you are an expert in both. The features in Oracle, SQL server, Teradata etc are a bit different and their syntaxes are different but structures are similar. Say for oracle, it is declare...begin(nesting begin end)....exception ...end;
Thanks Raja! I coundn't find any syntax examples for functions.
Can someone help me with this?
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3 October 2014
Post by James Doherty
Cover photo by Department of Foreign Affers
Reading time ~
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Nicolas Maire of the Swiss Tropical and Public Health Institute tells James Doherty how his team is using Crowdcrafting.org to help fight Malaria in Africa.
Rural Geolocator is a project hosted by Crowdcrafting which asks the public to identify houses in satellite images, currently of two locations in Africa: Rusinga Island in Lake Victoria, Kenya and Majete Wildlife Reserve in Malawi.
Field teams trying to stop the spread of Malaria need to deliver technologies and advice to indigenous populations to prevent them from contracting the disease. To deliver these tools, the teams need to know where people live. While they try their best to visit everyone, inevitably they miss some people out who, perhaps, live in remote areas which are difficult to access. Rural Geolocator helps to ensure that the field teams can visit everyone in a particular district.
I work for the Swiss Tropical and Public Health Institute, an associated institute of the University of Basel. We set up Health Demographic Surveillance Systems (HDSS) to intensively survey populations for which there is no routine registration of births and deaths. We try to enumerate these populations and understand the healthcare challenges they face.
Photo by Nicolas Maire HDSS enumerator team getting ready for a field survey in Majete, Malawi.
Teams investigating the effect of Malaria on populations in Africa realised that the data we collect would be really useful for their research – so they got in touch to ask for our help.
On Rusinga Island they install solar-powered mosquito traps outside people's houses. Odour baits within the traps mimic the smell of humans and catch mosquitoes before they go indoors. The intervention begins at one randomly selected house and expands radially until a cluster of houses with the intervention is created. Other clusters are created until all houses on the island have traps installed.
In Majete they use a variety of techniques, including sealing houses with insect screens, using biological agents to treat stagnant water which provides a breeding ground for mosquitoes, and using the 'roll-back' technique: A combination of mosquito nets and medication.
We need help to analyse thousands of satellite images (imported from Bing) to locate structures where people might live. There is a diverse range of possibilities of what constitutes a 'potentially inhabitable structure' that only the human brain can interpret. We couldn't use computer algorithms for these tasks.
An unexpected bonus of using Crowdcrafting has been the outreach achieved – lots more people know about the research projects because we've crowdsourced the data.
Photo by Nicolas Maire Geolocators at the Open Day of the Swiss TPH in June 2014.
Our volunteers have already invested a lot of time and effort collecting data sets for the Rusinga and Majete projects so the first thing to do is analyse them. We will create maps of potentially inhabitable structures to overlay maps of houses visited by the field teams which will help to identify missed households. Ultimately we hope that using satellite imagery and crowdsourcing will greatly improve our data to guide both field research and the implementation of malaria control measures.
Want to help field workers in Africa fight Malaria? Here's your chance: Rural Geolocator
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On our farm we grow good food.
We produce high quality meats, poultry and eggs, while keeping animal welfare, soil health, and sustainability as the highest priorities.
Local, free-range chicken, pork, lamb, and eggs
A signature line of gourmet fresh sausages you won't find anywhere else
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Find us at the City Market on 104th Street every Saturday from 9-3
See our list of products and pricing or shop online