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Adenoviral vector–mediated gene transfer of IL-13Rα2 chain followed by IL-13 cytotoxin treatment offers potent targeted therapy for cytotoxin-resistant cancers
Version of Record online: 8 MAR 2005
DOI: 10.1002/ijc.20995
Copyright © 2005 Wiley-Liss, Inc.
International Journal of Cancer
Volume 116, Issue 1, pages 1–8, 10 August 2005
Additional Information
How to Cite
Saito, M., Murata, T., Watanabe, K., Kawakami, K., Suzuki, M., Koji, T., Puri, R. K., Kitazato, K. and Kobayashi, N. (2005), Adenoviral vector–mediated gene transfer of IL-13Rα2 chain followed by IL-13 cytotoxin treatment offers potent targeted therapy for cytotoxin-resistant cancers. Int. J. Cancer, 116: 1–8. doi: 10.1002/ijc.20995
Publication History
- Issue online: 12 MAY 2005
- Version of Record online: 8 MAR 2005
- Manuscript Accepted: 22 DEC 2004
- Manuscript Received: 13 OCT 2004
Funded by
- Ministry of Education, Science, Technology, Sports and Culture of Japan
- Abstract
- Article
- References
- Cited By
- IL-13 cytotoxin;
- IL-13Rα2 chain;
- adenoviral vector;
- molecular targeting
Previous studies demonstrated that IL-13Rα2 chain–overexpressing cancer cells were highly sensitive to IL-13 cytotoxin (IL13-PE38QQR) and could be targeted by cytotoxin treatment. However, the majority of human tumors do not express high levels of IL-13Rα2 chain. To expand the IL-13 cytotoxin–mediated cancer targeting therapy, we combined cytotoxin treatment with gene transfer of IL-13Rα2 chain. We constructed a recombinant adenoviral vector carrying the human IL-13Rα2 gene (Ad-IL-13Rα2), which expresses high levels of IL-13Rα2 chain on infected cells. Human cancer cell lines A549 and HOS, which originally show no IL-13Rα2 expression and little sensitivity to IL-13 cytotoxin, were effectively converted to become sensitive to this cytotoxin after Ad-IL-13Rα2 infection. The CC50 of IL-13 cytotoxin for Ad-IL-13Rα2-infected A549 cells was <10 ng/ml, whereas the CC50 for uninfected or control vector-infected cells was >500 ng/ml. We also examined the antitumor activity of IL-13 cytotoxin in an established xenograft model of cytotoxin-resistant human lung tumor. Only a single i.t. injection of Ad-IL-13Rα2 markedly enhanced the sensitivity of established tumors to IL-13 cytotoxin treatment; furthermore, this antitumor effect was significantly sustained for more than 1 month after the last treatment with IL-13 cytotoxin. Taken together, these results suggest the combination of adenoviral vector–mediated IL-13Rα2 gene transfer and IL-13 cytotoxin administration can be an effective targeting approach for several types of IL-13 cytotoxin–resistant cancers which show no or little expression of IL-13Rα2 chain. © 2005 Wiley-Liss, Inc.
Targeted anticancer therapies have become increasingly successful in the treatment of various cancers. To limit nonspecific toxicity and to improve the efficiency of cancer therapy, tumor markers, which are generally overexpressed on the surface of tumor cells, can be selectively targeted. Previous studies of our and other groups have shown that IL-13Rα2 chain is one of the tumor-associated markers1, 2 that is overexpressed on several types of human cancer cell, such as RCC,3, 4, 5 glioma,4, 5, 6, 7, 8, 9, 10 AIDS-associated Kaposi's sarcoma,11, 12 head-and-neck cancer13, 14 and some ovarian cancers.4, 15 To establish a novel targeting therapy for such cancers, we had tried to target IL-13Rα2 chain by developing a potential therapeutic agent, IL-13 cytotoxin (IL13-PE38QQR),16 which is a recombinant fusion protein composed of IL-13 and a modified form of PE. Previous studies have proved that IL-13 cytotoxin is highly selective and potent in killing cancer cells that overexpress IL-13Rα2 chain.3, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 17 Preclinical studies regarding the safety and toxicity of IL-13 cytotoxin have been performed in mice, rats and monkeys; and all animals tolerated this therapy well, with minimal toxicity to vital organs.18 Based on these results, 3 phase I/II clinical trials of this cytotoxin in adults with malignant glioma have been initiated, and all are currently ongoing in the United States.18 However, the majority of human tumors do not express high levels of IL-13Rα2 chain;4, 14 thus, such tumor cells were practically resistant to this cytotoxin.14
IL-13 is a pleiotropic and Th2-dominated immune regulatory cytokine,19, 20 which plays a central role in allergic asthma21, 22, 23 and mediates diverse responses during helminth infection.24, 25 It is critical for tumor immunosurveillance26, 27 and modulates apoptosis or tumor cell growth.28, 29 Two different components of the IL-13 receptor have been identified, which are known as IL-13Rα1 chain30, 31, 32 and IL-13Rα2 chain.33, 34 IL-13Rα1 chain alone binds IL-13 with low affinity. Therefore, IL-13Rα1 chain requires the recruitment of IL-4Rα chain30, 31, 32, 35 to form a high-affinity heterodimer receptor complex, which mediates the IL-13 signal-transduction pathway. IL-13Rα2 chain by itself can bind IL-13 with higher affinity, but it does not mediate IL-13 signaling35, 36 because it has a very short cytoplasmic domain that lacks the conserved signaling motifs. In particular, IL-13Rα2 chain has a unique property of ligand internalization after binding,36, 37 which has also been identified as a soluble protein in mouse serum and urine;38 thus, it can be a specific inhibitor of IL-13 signaling or a decoy receptor for IL-13.39, 40, 41 Moreover, IL-13Rα2 chain may inhibit IL-13-mediated biologic functions in tumor growth and recurrence.26, 27, 42, 43
To utilize the attractive properties of IL-13Rα2 chain in IL-13 cytotoxin–mediated cancer therapy, we previously tried to combine the cytotoxin treatment with gene transfer of IL-13Rα2 chain.13, 14, 44, 45, 46 In the present study, we generated a replication-defective adenovirus, Ad-IL-13Rα2, which can express high levels of IL-13Rα2 chain in infected cells. We confirmed that adenovirus-mediated gene transfer of IL-13Rα2 chain significantly enhanced the in vitro cytotoxic effect of IL-13 cytotoxin in cytotoxin-resistant cancer cells and the in vivo antitumor activity of the cytotoxin in an established human tumor model which is resistant to the cytotoxin. Our results demonstrate that combination therapy of adenoviral gene transfer of IL-13Rα2 chain and IL-13 cytotoxin treatment can offer a potent targeting approach to cancer therapy.
Material and methods
Recombinant cytokines and immunotoxin
Recombinant human IL-4 and IL-13 were purchased from Peprotech (London, UK). Recombinant IL-13 cytotoxin (IL13-PE38QQR) was produced and purified as previously described.14
Cell lines
A549 (human lung cancer), PM-RCC (renal cell carcinoma) and 293 (human embryonic kidney) cell lines were cultured in DMEM (Sigma, St. Louis, MO) containing 10% FBS, 100 units/ml penicillin and 100 μg/ml streptomycin (Nacalai Tesque, Kyoto, Japan). Human osteosarcoma cell lines HOS and MG-63 were cultured in Eagle's minimum essential medium (Sigma) containing 10% FBS, 100 units/ml penicillin and 100 μg/ml streptomycin.
Construction of replication-defective ΔE1ΔE3 adenoviral vector
A replication-defective ΔE1ΔE3 adenoviral vector expressing the IL-13Rα2 chain gene under CMV promoter, which was termed Ad-IL-13Rα2, was constructed as described previously.47 The human IL-13Rα2 chain cDNA fragment was inserted into the NotI site of shuttle plasmid pCMV-SV2+,48 which contains a portion of the adenovirus type 5 (Ad5) genome and the CMV early promoter/enhancer. The shuttle plasmid carrying the expression cassette of IL-13Rα2 chain instead of the Ad5 E1 region and adenoviral genomic plasmid pJM17 (Microbix Biosystems, Toronto, Canada) containing the Ad5 genome with E1 and E3 deletions were cotransfected into 293 cells using the Calcium-Phosphate Transfection System (Life Technologies, Grand Island, NY). By homologous recombination between the above 2 plasmids in 293 cells, cytopathic effects were observed within 2 weeks after transfection, after which cells were harvested. Viruses were released from cells by freeze-thawing using liquid N2. Another replication-defective adenoviral vector expressing the β-galactosidase gene, named Ad-LacZ, was constructed in the same way and utilized as an adenoviral infection control. Both adenoviral vectors were purified from a single plaque clone, propagated in 293 cells, and concentrated by the cesium chloride-density gradient centrifugation method followed by dialysis. Stocks of recombinant viruses were prepared and stored at –80°C until use. The titer of recombinant virus stocks was checked on 293 cells by a plaque-forming assay.
Conditions of recombinant adenoviral vector infection
Cells were plated in 6-well (1.0 × 105/well) or 24-well (2.0 × 104/well) culture plates and incubated at 37°C in a humidified 5% CO2 incubator for 24 hr. Cells were infected with purified adenoviral vector at several concentrations in 800 μl (6-well plate) or 200 μl (24-well plate) of serum-free medium at 37°C in a humidified 5% CO2 incubator for 2 hr. Then, fresh complete medium was added to each well.
RT-PCR analysis
Total RNA of cells was extracted using Trizol reagent (Life Technologies). Each 2.5 μg of the RNA samples was used for an RT reaction. The reaction was performed at 37°C for 50 min in a final volume of 20 μl of reaction mixture containing 50 mM TRIS-HCl (pH 8.3), 3 mM MgCl2, 75 mM KCl, 0.5 mM of each dNTP, 40 units of RNase inhibitor (Nacalai Tesque), 500 ng of oligo-d(T)20 primer and 200 units of MMLV reverse transcriptase (Life Technologies). Each 1 μl aliquot of the cDNA samples was amplified in a final volume of 50 μl of PCR mixture containing 10 mM TRIS-HCl (pH 8.8), 2 mM MgCl2, 50 mM KCl, 0.2 mM of each dNTP, 2.5 units of SuperTaq DNA polymerase (Ambion, Austin, TX) and 0.2 μM each of specific primers for human IL-13Rα2 chain cDNA (5′-AATGGCTTTCGTTTGCTTGG-3′, 5′-ACGCAATCCATATCCTGAAC-3′, in Fig. 1; 5′-ATGGCTTTCGTTTGCTTGG-3′, 5′-CATGTATCACAGAAAAATT-3′ in Fig. 2), human IL-13Rα1 chain cDNA (5′-TATTTTAGTCATTTTGGCGACAAAC-3′, 5′-GTTATGTAGAGTGTGGAATTGCGCT-3′) or human IL-4Rα chain cDNA (5′-GACCTGGAGCAACCCGTATC-3′, 5′-CATAGCACAACAGGCAGACG-3′). An internal control experiment in RT-PCR analysis was performed using a human β-actin primer pair (R&D Systems, Minneapolis, MN) or GAPDH-specific primers (5′-ACCACAGTCCATGCCATCAC-3′, 5′-TCCACCACCCTGTTGCTGTA-3′). PCR was carried out for 30 cycles at 95°C for 30 sec, 55°C for 30 sec and 72°C for 60 sec. PCR product was run in a 1% agarose gel. The intensities of each amplified IL-13Rα2 cDNA band were analyzed by Scion Image software (version 4.02; Scion, Frederick, MD) and exhibited as values relative to internal control, β-actin or GAPDH.
Flow cytometry
A549 and HOS cells were infected with Ad-IL-13Rα2 at MOI 1–100 and, after 48 hr, harvested and incubated with FITC-conjugated mouse MAb against human IL-13Rα2 chain (1:100 dilution; Diaclone, Besançon, France) or FITC-conjugated mouse IgG1 isotype-matched control antibody (1:100 dilution; Dako Cytomation, Glostrup, Denmark) in the dark on ice for 2 hr. FITC-labeled cells were analyzed by a Coulter EPICS-XL flow cytometer (System II software; Beckman Coulter, Miami, FL).
Evaluation of sensitivity to IL-13 cytotoxin on cancer cell lines
Cancer cell lines (A549, HOS, MG-63 and PM-RCC) were treated with IL13-PE38QQR (final concentration 100 ng/ml) at 37°C in a humidified 5% CO2 incubator for 72 hr, washed and harvested. The cytotoxic effect of IL13-PE38QQR was evaluated by counting viable cells using trypan blue dye exclusion and exhibited as a value relative to that of nontreated control.
Analysis of the cytotoxic effect of IL-13 cytotoxin on adenoviral vector–infected cells
A549 and HOS cells were infected with either Ad-IL-13Rα2 or Ad-LacZ at different concentrations (MOI 0–100) followed by IL13-PE38QQR (100 ng/ml) treatment for 72 hr or infected with adenoviral vectors at MOI 10 followed by treatment with different concentrations of IL13-PE38QQR (0–500 ng/ml) for 72 hr. Cell viability was determined and exhibited as a value relative to that of nontreated control.
In vitro proliferation assay
A549 cells were infected with Ad-IL-13Rα2 or Ad-LacZ at MOI 10 and treated with IL13-PE38QQR (100 ng/ml every 3 days) for 14 days. The number of living cells was assessed on the indicated days (day 3, 7, 11 and 14).
Competitive effect of IL-13 on cytotoxicity of IL-13 cytotoxin
A549 cells were infected with Ad-IL-13Rα2 or Ad-LacZ at MOI 10 followed by IL13-PE38QQR (10 ng/ml) treatment and incubated in the presence or absence of recombinant human IL-13 or IL-4 (500 ng/ml) for 72 hr.
Human lung cancer xenograft model and treatment protocol
Four-week-old nude mice, BALB/cA jcl-nu/nu (Clea, Tokyo, Japan), were housed in sterilized filter-topped cages and maintained in a pathogen-free condition. Animal care was in accordance with the guidelines of the Nagasaki University Animal Research Advisory Committee. Tumorigenic human lung cancer A549 cells (5.0 × 106/body) were injected s.c. into the dorsal surface of mice, and that day was appointed as day 0 after transplantation of tumors. Palpable tumors developed within 5–7 days. Mice bearing A549 tumors received i.t. injection of 1.0 × 109 pfu/body of Ad-IL-13Rα2 or Ad-LacZ on day 5 and i.t. injection of IL13-PE38QQR (250 μg/kg/day) or PBS (excipient) on days 7, 8, 9, 10 and 11 using a 25 μl microsyringe (Hamilton, Reno, NV) with a 26-gauge needle. Tumor size was measured by caliper every 3 days. Approximate tumor size was calculated as the product of the 2 perpendicular diameters.
Statistical analysis
Data were presented as means ± SEM or SD. Differences between 2 groups were compared using 2-tailed Student's t-test. One-way ANOVA was used to examine differences between the multiple treatment groups in competition assay for the cytotoxic effect of IL-13 cytotoxin and in the tumor size measurement experiment. When ANOVA indicated a significant difference (p < 0.05), a post hoc multiple comparison test was performed using Dunnett's 2-tailed t-test or Tukey's HSD test. Differences were considered statistically significant at p < 0.05.
IL-13Rα2 chain expression and sensitivity to IL-13 cytotoxin in various cancer cell lines
To confirm the correlation between expression of IL-13Rα2 chain and sensitivity to IL-13 cytotoxin (IL13-PE38QQR) treatment in cells, several human cancer cell lines (A549, HOS, MG-63 and PM-RCC) were examined for mRNA expression of IL-13Rα2 chain by RT-PCR analysis and for sensitivity to IL-13 cytotoxin. As shown in Figure 1a, all of these cancer cell lines expressed mRNA of both IL-4Rα and IL-13Rα1 at equivalent levels, whereas expression of IL-13Rα2 mRNA was observed only in MG-63 and PM-RCC cells but not in A549 and HOS cells. In MG-63 and PM-RCC cells, treatment with IL-13 cytotoxin (final concentration 100 ng/ml) actually showed 70–80% of the cytotoxicity (Fig. 1b). By contrast, in both IL-13Rα2 mRNA-negative cell lines, A549 and HOS, IL-13 cytotoxin treatment showed <5% of the cytotoxicity, regardless of mRNA expression of IL-13Rα1 and IL-4Rα (Fig. 1a). In addition, we found that the other human cancer cell lines, Caco-2 (colon cancer), PANC-1 (pancreatic cancer) and MDA-MB-231 (breast cancer), expressing mRNA of IL-4Rα and IL-13Rα1 but not IL-13Rα2, were similarly resistant to IL-13 cytotoxin (data not shown). These results suggest that the cytotoxicity of IL-13 cytotoxin to these cancer cell lines is dependent on the expression of IL-13Rα2 chain and independent of expression of IL-13Rα1 chain and IL-4Rα chain.
IL-13Rα2 chain expression in cancer cells provided by adenoviral vector–mediated gene transfer
For the purpose of IL-13Rα2 chain delivery to target cancer cells, we generated a recombinant replication-defective adenoviral vector, Ad-IL-13Rα2.
To confirm the expression of IL-13Rα2 mRNA by Ad-IL-13Rα2 infection, A549 and HOS cells, which showed no expression of IL-13Rα2 chain (Fig. 1a), were infected with Ad-IL-13Rα2 at different infection-doses (MOI 1, 10), and mRNA expression was detected by RT-PCR. As shown in Figure 2a, expression of IL-13Rα2 mRNA in both A549 and HOS cells was detected within 24 hr after Ad-IL-13Rα2 infection at MOI 10 and at 48 hr after MOI 1 infection. The high level of IL-13Rα2 mRNA expression was sustained for 7 days after infection at MOI 1 and for 14 days at MOI 10 in A549 cells (data not shown).
To confirm the expression of IL-13Rα2 protein on the surface of Ad-IL-13Rα2-infected cells, we performed flow cytometry using FITC-conjugated antibody against human IL-13Rα2 chain. As shown in Figure 2b, expression of IL-13Rα2 chain was detected at the surface of both A549 and HOS cells infected with Ad-IL-13Rα2 vector in a dose-dependent manner.
Both RT-PCR and flow-cytometric data showed that expression of IL-13Rα2 chain on A549 cells was 2- to 4-fold higher than that on HOS cells (Fig. 2a, ratio of cDNA band intensity and Table I). We also evaluated the efficiency of adenoviral vector–based gene transfer in these cell lines using Ad-LacZ, which expresses the β-galactosidase gene, and observed that A549 cells were more susceptible to adenoviral infection compared to HOS cells (data not shown). Thus, the difference in adenovirus-mediated gene expression between these cell lines appears to be consistent with the susceptibility to adenoviral infection.
|Cell line/treatment
|Isotype control
|Ad-IL-13Rα2 MOI 1
|Ad-IL-13Rα2 MOI 10
|Ad-IL-13Rα2 MOI 100
|Ad-IL-13Rα2 MOI 1
|Ad-IL-13Rα2 MOI 10
|Ad-IL-13Rα2 MOI 100
In vitro IL-13Rα2 gene transfer by Ad-IL-13Rα2 augments sensitivity of cancer cells to IL-13 cytotoxin
To investigate an effect of Ad-IL-13Rα2 vector on the sensitivity of the target cancer cells to IL-13 cytotoxin, cytotoxin-resistant cancer cells A549 and HOS (Fig. 1b) were infected with Ad-IL-13Rα2 at different infection-doses (MOI 0–100) followed by treatment with IL-13 cytotoxin (100 ng/ml) for 72 hr and the number of viable cells determined. As shown in Figure 3a,b, both A549 and HOS cells infected with Ad-IL-13Rα2 vector at different infection-doses were effectively killed by IL-13 cytotoxin in a dose-dependent manner, whereas control cells which were uninfected or infected with Ad-LacZ vector did not respond to IL-13 cytotoxin at all. The sensitivity of Ad-IL-13Rα2-infected A549 cells to IL-13 cytotoxin was relatively higher than that of Ad-IL-13Rα2-infected HOS cells, which correlated with the difference in IL-13Rα2 chain expression between these 2 types of cancer cell after Ad-IL-13Rα2 infection (Fig. 2a,b).
Furthermore, we examined the dose effect of IL-13 cytotoxin (10–500 ng/ml for 72 hr) on both A549 and HOS cells infected with Ad-IL-13Rα2 at MOI 10 (Fig. 3c). When Ad-IL-13Rα2-infected A549 cells were treated with IL-13 cytotoxin at doses above 50 ng/ml, >80% of them were killed. The CC50 (Table II) of IL-13 cytotoxin for Ad-IL-13Rα2-infected A549 cells was <10 ng/ml, which was much more highly sensitive than that of PM-RCC (CC50 approx. 60 ng/ml). The viability of HOS cells infected with Ad-IL-13Rα2 was also reduced by IL-13 cytotoxin treatment in a dose-dependent manner (Fig. 3c, CC50 almost 200 ng/ml). However, both A549 and HOS cells after no infection or infection with Ad-LacZ vector were not sensitive to IL-13 cytotoxin (CC50 >500 ng/ml).
|Cell line/treatment
|CC50 (ng/ml)
|59.1 ± 8.4
|> 500
|Ad-LacZ MOI 10
|> 500
|Ad-IL-13Rα2 MOI 10
|7.1 ± 3.2
|> 500
|Ad-LacZ MOI 10
|> 500
|Ad-IL-13Rα2 MOI 10
|208.8 ± 15.6
We also examined the long-term effect of IL-13 cytotoxin treatment on the growth of target cells infected with Ad-IL-13Rα2. A549 cells were infected with Ad-IL-13Rα2 at MOI 10 and incubated in the presence of IL-13 cytotoxin (100 ng/ml) for 14 days. As shown in Figure 3d, A549 cells without adenoviral vector infection demonstrated vigorous tumor cell proliferation regardless of treatment with IL-13 cytotoxin. Ad-IL-13Rα2-infected cells hardly proliferated for the first 7 days despite the absence of treatment with IL-13 cytotoxin; but afterward, cells grew linearly in a similar fashion to untreated/uninfected cells. A similar tendency was also observed in Ad-LacZ-infected cells. The number of Ad-IL-13Rα2-infected cells evidently decreased along with IL-13 cytotoxin treatment, and almost all cells were killed within 3–7 days after initiation of the combination treatment.
IL-13, but not IL-4, competitively inhibits cytotoxicity of IL-13 cytotoxin in cancer cells infected with Ad-IL-13Rα2
IL-13 cytotoxin, which is a derivation of IL-13, can also bind to 2 types of receptor, physiologic IL-13Rα1/IL-4Rα and decoy IL-13Rα2. Previous studies have demonstrated that the attachment of radiolabeled IL-13 to the functional heterodimer receptor can be competed by IL-4 as well as IL-13,30, 31, 32, 49 whereas its attachment to IL-13Rα2 is competed by IL-13 but not IL-4.2, 9, 10, 15, 33, 36 To confirm that the cytotoxicity of IL-13 cytotoxin was specifically mediated through IL-13Rα2 chain delivered by Ad-IL-13Rα2 vector, A549 cells were infected with Ad-IL-13Rα2 at MOI 10 followed by treatment with IL-13 cytotoxin (10 ng/ml) in the presence of a 50-fold dose of human IL-13 or IL-4. As shown in Figure 4, IL-13 and IL-4 did not affect the growth of uninfected or Ad-LacZ-infected cells, regardless of treatment with IL-13 cytotoxin. In contrast, IL-13 cytotoxin was significantly cytotoxic to only Ad-IL-13Rα2-infected cells (p < 0.05); furthermore, the cytotoxic effect was completely abolished by a 50-fold amount of IL-13 (p < 0.01) but not the same amount of IL-4. These results strongly suggest that the cytotoxic effects of IL-13 cytotoxin are selectively mediated through IL-13Rα2 chain but not the functional receptor of the IL-13Rα1/IL-4Rα heterodimer complex.
Administration i.t. of Ad-IL-13Rα2 vector followed by IL-13 cytotoxin leads to significant regression and sustained growth inhibition of A549 tumor xenografts in nude mice
To confirm the combination effect of Ad-IL-13Rα2 infection and IL-13 cytotoxin treatment in vivo, we established a mouse xenograft model of the cytotoxin-resistant human tumor. A549 cells were s.c.-implanted in nude mice (day 0), and then the established tumors received i.t. injection of Ad-IL-13Rα2 or Ad-LacZ on day 5. Subsequently, IL-13 cytotoxin (250 μg/kg/day) or PBS was administered to the tumors on a daily basis for 5 days from day 7 through day 11. From adenoviral vector injection (day 5) to initiation of IL-13 cytotoxin treatment (day 7), no significant differences in the size of tumors between all experimental groups were observed (p > 0.1, one-way ANOVA), regardless of different adenoviral vector injections. However, as shown in Figure 5a,b, tumors treated with i.t. administration of Ad-IL-13Rα2 followed by IL-13 cytotoxin significantly showed immediate regression and sustained stationary growth (p < 0.05 vs. control groups from day 11 to day 54 after implantation) for 43 days. At the end of a 2-month course of i.t. combined treatment with Ad-IL-13Rα2 and IL-13 cytotoxin (on day 54), tumor size was approximately 70% less than that of control groups (p < 0.01). However, tumor size after Ad-LacZ injection followed by IL-13 cytotoxin treatment showed no statistical difference compared to tumors injected with Ad-LacZ followed by excipient injection (p > 0.1). Among all treated mice, noticeable histologic changes or apparent side effects were not observed in nontargeted tissues such as liver and kidneys; moreover, macroscopically detectable toxic signs including loss of body weight were not noticed either (data not shown).
We first demonstrated that delivery of IL-13Rα2 chain using adenoviral vector to IL-13 cytotoxin-resistant cancer cells (Fig. 2) remarkably enhanced the sensitivity of these cells to IL-13 cytotoxin in vitro (Figs. 3, 4). The cytotoxic activity of IL-13 cytotoxin to cancer cells appears to specifically require expression of IL-13Rα2 chain because IL-13Rα2-negative cells hardly showed any response to IL-13 cytotoxin treatment despite expression of both IL-13Rα1 and IL-4Rα (Fig. 1), and excess dose of IL-13, but not IL-4, significantly abolished the cytotoxic effect of IL-13 cytotoxin on Ad-IL-13Rα2-infected cancer cells (Fig. 4). Furthermore, we demonstrated that in vivo i.t. administration of Ad-IL-13Rα2 vector markedly enhanced the antitumor activity of IL-13 cytotoxin in the cytotoxin-resistant human tumor xenograft model (Fig. 5). These results are consistent with our previous findings, in which plasmid-based gene transfer of IL-13Rα2 chain showed enhanced sensitivity of tumor cells to IL-13 cytotoxin treatment.13, 14, 44, 45, 46 In the present study, only one injection of Ad-IL-13Rα2 was sufficient for IL-13 cytotoxin treatment to cause immediate and significant regression of these established tumors and sustained growth inhibition for more than 1 month after the last administration of IL-13 cytotoxin. Both in vitro and in vivo experiments demonstrated that the antitumor activity of combined treatment with Ad-IL-13Rα2 and IL-13 cytotoxin on the target cancer cells or tumors was independent of the toxic effects derived from adenoviral vector itself (Figs. 3–5).
The i.t. administration of IL-13 cytotoxin, even at high daily doses (250 μg/kg for 5 days), did not show any nonspecific toxic effects on tumors infected with the control vector (Fig. 5). Our previous studies have already confirmed that i.p. or i.v. approaches of IL-13 cytotoxin can also kill tumors expressing IL-13Rα2 chain8, 11, 13, 50, 51 without any nonspecific toxic signs;50, 51 thus, this cytotoxin should not cause nonspecific side effects or collateral damage to nontargeted normal cells. Moreover, our previous study also showed that the half-life of IL-13 cytotoxin in the blood is relatively short.11 If any serious side effect was caused by IL-13 cytotoxin, there is a possibility that it would be readily controlled by adjustment of the toxin dose even after gene transfer of IL-13Rα2 chain. Except for a certain kind of cancer cells,43 IL-13Rα2 chain by itself is not cytotoxic to targeted cancer cells, even though it is overexpressed on cells after gene transfer; this was confirmed in our data (Figs. 2–5). These findings support the contention that the high-level expression of IL-13Rα2 chain using our adenoviral vector on the surface of targeted cancer cells is not only a safe procedure but also a promising strategy for cancer targeting therapy using IL-13 cytotoxin, which itself is safe and specific.
Our previous study showed that in vivo overexpression of IL-13Rα2 chain inhibited tumorigenicity of pancreatic and breast cancer cell lines in nude mice.42 The mechanism of such antitumorigenic activity of IL-13Rα2 chain was attributed to infiltration of inflammatory cells, including neutrophils, and production of cytokines.42 In the end of our in vivo experiment, histologic analysis also showed that prominent infiltration of inflammatory cells was indeed induced in only Ad-IL-13Rα2-infected tumor tissues, whereas tumor tissues obtained from mice injected with Ad-IL-13Rα2 followed by IL-13 cytotoxin treatment were free from inflammatory cells (data not shown). Such infiltration of inflammatory cells was specifically observed in tumors administered Ad-IL-13Rα2 vector but not Ad-LacZ control vector (data not shown); thus, it appeared to be relatively a long-term and IL-13Rα2 chain–specific phenomenon, not a distinctive inflammatory response caused by adenoviral vector itself, because it had been observed nearly 2 months after administration of adenoviral vectors. In addition, a soluble form of IL-13Rα2 can prevent IL-13-mediated suppression of tumor immunosurveillance.26, 27 Therefore, i.t. introduction of IL-13Rα2 chain into established tumors may efficiently lead to activation of an antitumorigenic response and tumor immunosurveillance in tumor tissues.
To realize clinical applications of our combination approach based on in vivo injection of Ad-IL-13Rα2 vector, we need to achieve not only high efficiency but also more specific gene transfer and expression to target tumor cells because adenovirus-based vectors can infect a broad variety of mammalian cell types, not only cancer cells but also normal cells. Previously, it was reported that use of promoter and expression regulatory elements of tumor- or tissue-specific molecules, such as α-fetoprotein, carcinoembryonic antigen, human telomerase reverse transcriptase and prostate-specific antigen, successfully regulated the expression of transgenes in a tumor- or tissue-specific fashion after systemic gene transfer using adenoviral vector.52, 53, 54, 55 Therefore, these targeting techniques of adenovirus-based vectors will further improve our combination strategy.
In conclusion, our studies prove that adenoviral vector–mediated gene delivery of IL-13Rα2 chain combined with IL-13 cytotoxin treatment effectively and selectively kill targeted cancer cells in vitro and in vivo and suggest that our combined strategy offers a potent targeting approach for cancer treatment.
We thank Dr. B.H. Joshi (Laboratory of Molecular Tumor Biology, Center for Biologics Evaluation and Research, FDA) for providing purified recombinant IL13-PE38QQR; Dr. Y. Hishikawa and Ms. M. Kawakatsu (Laboratory of Histology and Cell Biology, Nagasaki University) for histologic analysis; and Dr. Y. Niu, Ms. Y. Ma, Ms. N. Okamoto, Ms. R. Takashima, Ms. I. Fujii and Mr. H.F. Rafidinarivo (Laboratory of Molecular Biology of Infectious Agents, Nagasaki University) for technical assistance. We also thank Dr. M. Kawakami (Laboratory of Molecular Tumor Biology, Center for Biologics Evaluation and Research, FDA) for critical reading of this manuscript. This work was supported in part by a grant-in-aid from the Ministry of Education, Science, Technology, Sports and Culture of Japan (to T.M.).
- 1Interleukin-13 receptor α2 chain in human head and neck cancer serves as a unique diagnostic marker. Clin Cancer Res 2003; 9: 6381–8., , , , , , .
- 2Receptor for interleukin 13 is a marker and therapeutic target for human high-grade gliomas. Clin Cancer Res 1999; 5: 985–90., , , , .
- 3Targeting of interleukin-13 receptor on human renal cell carcinoma cells by a recombinant chimeric protein composed of interleukin-13 and a truncated form of Pseudomonas exotoxin A (PE38QQR). Blood 1996; 87: 4333–9., , , , , , , .
- 4Structure of IL-13 receptor: analysis of subunit composition in cancer and immune cells. Biochem Biophys Res Commun 1997; 238: 90–4., , , .
- 5Expression of interleukin 13 receptor in glioma and renal cell carcinoma: IL13Rα2 as a decoy receptor for IL13. Lab Invest 2001; 81: 1223–31., , , , , , , , .
- 6Human glioma cells overexpress receptors for interleukin 13 and are extremely sensitive to a novel chimeric protein composed of interleukin 13 and Pseudomonas exotoxin. Clin Cancer Res 1995; 1: 1253–8., , , , .
- 7Interleukin-13 receptor α chain: a novel tumor-associated transmembrane protein in primary explants of human malignant gliomas. Cancer Res 2000; 60: 1168–72., , .
- 8Interleukin-13 receptor as a unique target for anti-glioblastoma therapy. Int J Cancer 2001; 92: 168–75., , .
- 9Receptor for interleukin (IL) 13 does not interact with IL-4 but receptor for IL-4 interacts with IL-13 on human glioma cells. J Biol Chem 1996; 271: 22428–33., , , , .
- 10IL-13Rα2 is a glioma-restricted receptor for interleukin-13. Neoplasia 2002; 4: 388–99., , , , .
- 11Interleukin-13 fusion cytotoxin as a potent targeted agent for AIDS-Kaposi's sarcoma xenograft. Blood 2000; 95: 3506–13., .
- 12Receptor for interleukin 13 on AIDS-associated Kaposi's sarcoma cells serves as a new target for a potent Pseudomonas exotoxin–based chimeric toxin protein. Clin Cancer Res 1997; 3: 151–6., , , , , , .
- 13Interleukin-13 receptor–targeted cancer therapy in an immunodeficient animal model of human head and neck cancer. Cancer Res 2001; 61: 6194–200., , , .
- 14Heterogeneity in interleukin-13 receptor expression and subunit structure in squamous cell carcinoma of head and neck: differential sensitivity to chimeric fusion proteins comprised of interleukin-13 and a mutated form of Pseudomonas exotoxin. Clin Cancer Res 2002; 8: 1948–56., , , .
- 15Human ovarian carcinoma cell lines express IL-4 and IL-13 receptors: comparison between IL-4- and IL-13-induced signal transduction. Int J Cancer 1997; 70: 230–40., , , .
- 16A novel chimeric protein composed of interleukin 13 and Pseudomonas exotoxin is highly cytotoxic to human carcinoma cells expressing receptors for interleukin 13 and interleukin 4. J Biol Chem 1995; 270: 16775–80., , , .
- 17Interleukin-13 receptors on human prostate carcinoma cell lines represent a novel target for a chimeric protein composed of IL-13 and a mutated form of Pseudomonas exotoxin. J Urol 1997; 158: 948–53., , , , , , , , , , .
- 18Interleukin-13 receptor–directed cytotoxin for malignant glioma therapy: from bench to bedside. J Neurooncol 2003; 65: 37–48., .
- 19Interleukin-13 is a new human lymphokine regulating inflammatory and immune responses. Nature 1993; 362: 248–50., , , , , , , , , , , , et al.
- 20IL-13 effector functions. Annu Rev Immunol 2003; 21: 425–56..
- 21Interleukin-13: central mediator of allergic asthma. Science 1998; 282: 2258–61., , , , , , .
- 22Requirement for IL-13 independently of IL-4 in experimental asthma. Science 1998; 282: 2261–3., , , , , , , , , , .
- 23Direct effects of interleukin-13 on epithelial cells cause airway hyperreactivity and mucus overproduction in asthma. Nat Med 2002; 8: 885–9., , , , , , , , .
- 24Schistosome infection of transgenic mice defines distinct and contrasting pathogenic roles for IL-4 and IL-13: IL-13 is a profibrotic agent. J Immunol 2000; 164: 2585–91., , , .
- 25A critical role for IL-13 in resistance to intestinal nematode infection. J Immunol 1998; 160: 3453–61., , .
- 26NKT cell-mediated repression of tumour immunosurveillance by IL-13 and the IL-4R-STAT6 pathway. Nat Immunol 2000; 1: 515–20., , , , , , , , .
- 27Unmasking immunosurveillance against a syngeneic colon cancer by elimination of CD4+ NKT regulatory cells and IL-13. Int J Cancer 2005; 114: 80–7., , , , .
- 28Interleukin-13 inhibits interleukin-2-induced proliferation and protects chronic lymphocytic leukemia B cells from in vitro apoptosis. Blood 1996; 87: 1022–9., , , , , , , , , .
- 29Interleukin 13 is secreted by and stimulates the growth of Hodgkin and Reed-Sternberg cells. J Exp Med 1999; 189: 1939–46., , , , , , , , , , , , et al.
- 30Cloning and characterization of a binding subunit of the interleukin 13 receptor that is also a component of the interleukin 4 receptor. Proc Natl Acad Sci USA 1996; 93: 497–501., , , , , .
- 31cDNA cloning and characterization of the human interleukin 13 receptor α chain. J Biol Chem 1996; 271: 29265–70., , , , , .
- 32Cloning of the human IL-13Rα1 chain and reconstitution with the IL-4Rα of a functional IL-4/IL-13 receptor complex. FEBS Lett 1997; 401: 163–6., , , , , , .
- 33Cloning and characterization of a specific interleukin (IL)-13 binding protein structurally related to the IL-5 receptor α chain. J Biol Chem 1996; 271: 16921–6., , , , , , .
- 34The murine IL-13 receptor α2: molecular cloning, characterization, and comparison with murine IL-13 receptor α1. J Immunol 1998; 161: 2317–24., , , , , , , , , , .
- 35Interleukin-13 receptor α′ but not α chain: a functional component of interleukin-4 receptors. Blood 1998; 91: 3884–91., , .
- 36The interleukin-13 receptor α2 chain: an essential component for binding and internalization but not for interleukin-13-induced signal transduction through the STAT6 pathway. Blood 2001; 97: 2673–9., , , .
- 37Identification of distinct roles for a dileucine and a tyrosine internalization motif in the interleukin (IL)-13 binding component IL-13 receptor α2 chain. J Biol Chem 2001; 276: 25114–20., , .
- 38Identification, purification, and characterization of a soluble interleukin (IL)-13-binding protein. Evidence that it is distinct from the cloned Il-13 receptor and Il-4 receptor α-chains. J Biol Chem 1997; 272: 9474–80., , , , , , , , , .
- 39The interleukin-4/interleukin-13 receptor of human synovial fibroblasts: overexpression of the nonsignaling interleukin-13 receptor α2. Lab Invest 1998; 78: 591–602., , , , , , , , , , , .
- 40Enhanced interleukin (IL)-13 responses in mice lacking IL-13 receptor α2. J Exp Med 2003; 197: 703–9., , , , , , , , , , , , et al.
- 41Regulation and function of the interleukin 13 receptor α2 during a T helper cell type 2-dominant immune response. J Exp Med 2003; 197: 687–701., , , , , , , , , , .
- 42In vivo overexpression of IL-13 receptor α2 chain inhibits tumorigenicity of human breast and pancreatic tumors in immunodeficient mice. J Exp Med 2001; 194: 1743–54., , , , .
- 43Soluble interleukin-13Rα2 decoy receptor inhibits Hodgkin's lymphoma growth in vitro and in vivo. Cancer Res 2004; 64: 3271–5., , , , , , , , , , , .
- 44Sensitization of cancer cells to interleukin 13-Pseudomonas exotoxin–induced cell death by gene transfer of interleukin 13 receptor α chain. Hum Gene Ther 2000; 11: 1829–35., , .
- 45Gene transfer of interleukin 13 receptor α2 chain dramatically enhances the antitumor effect of IL-13 receptor–targeted cytotoxin in human prostate cancer xenografts. Cancer Gene Ther 2001; 8: 861–8., , , .
- 46Interleukin-13 receptor-targeted cytotoxin cancer therapy leads to complete eradication of tumors with the aid of phagocytic cells in nude mice model of human cancer. J Immunol 2002; 169: 7119–26., , .
- 47Immune responses against replication-deficient adenovirus inhibit ovalbumin-specific allergic reactions in mice. Hum Gene Ther 2000; 11: 827–38., , , , , , , .
- 48In vitro and in vivo transfer and expression of human surfactant SP-A- and SP-B-associated protein cDNAs mediated by replication-deficient, recombinant adenoviral vectors. Hum Gene Ther 1995; 6: 277–87., , .
- 49The IL-13 receptor structure differs on various cell types and may share more than one component with IL-4 receptor. J Immunol 1997; 158: 756–64., , , , .
- 50Improved anti-tumor activity and safety of interleukin-13 receptor targeted cytotoxin by systemic continuous administration in head and neck cancer xenograft model. Mol Med 2002; 8: 487–94., , , .
- 51Specifically targeted killing of interleukin-13 (IL-13) receptor–expressing breast cancer by IL-13 fusion cytotoxin in animal model of human disease. Mol Cancer Ther 2004; 3: 137–47., , .
- 52Insulation from viral transcriptional regulatory elements enables improvement to hepatoma-specific gene expression from adenovirus vectors. Biochem Biophys Res Commun 2003; 307: 759–64., , , , , , , , .
- 53Regional delivery and selective expression of a high-activity yeast cytosine deaminase in an intrahepatic colon cancer model. Cancer Res 2003; 63: 658–63., , , , , , , .
- 54Chimeric PSA enhancers exhibit augmented activity in prostate cancer gene therapy vectors. Gene Ther 2001; 8: 1416–26., , , , , , .
- 55Tumor-specific gene therapy for undifferentiated thyroid carcinoma utilizing the telomerase reverse transcriptase promoter. J Clin Endocrinol Metab 2003; 88: 3531–8., , , , , , , , , , .


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StartUpHire is the place to find VP, Corporate Marketing jobs. Review the list of VP, Corporate Marketing career opportunities to the left. These VP, Corporate Marketing jobs represent some of the most rewarding careers available.
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at PulsePoint New York About You: Reporting to the Chief Operating Officer and leading a team of four, the (S)VP of Marketing will define, drive and implement a cohesive Corporate Marketing Strategy to position PulsePoint as THE leading global programmatic platform for brand5/27/2016 New York, NY Save This Job
Marketing Alameda, California Description Position at 24/7 Sr. Director, Web Marketing 247 is a rapidly growing Sequoia Capital-backed company that is transforming how the world's leading companies interact with consumers. 247 is an enterprise SaaS offering that enables4/26/2016 Alameda, CA Save This Job
Director, Product MarketingCompany Overview:Velocify is one of the fastest growing software-as-a-service companies in the country, providing the market-leading solution for intelligent sales automation and cloud-based telephony. Velocifyâs software is designed for6/12/2016 El Segundo, CA Save This Job
Reporting to the President, Tripwire seeks an experienced, strategic Vice President of Marketing. S/he will actively participate in business strategy with the executive management team. The VP of Marketing will own and drive all activities related to driving awareness and5/25/2016 Portland, OR Save This Job
Location: MBI Headquarters, Davis, CA Reports to: Vice President of Marketing Summary Headquartered in Davis, CA, Marrone Bio Innovations (MBI) (NASDAQ: MBII) is a global provider of bio-based pest management and plant health products for the agriculture, turf and ornamental,5/22/2016 Davis, CA Save This Job
Marketing New York, NY, United States Director/ VP of B2B Content & Communications Marketing xAd enables real-time, location-based marketing, without the guesswork. Working with xAd, marketers can deliver more relevant, personalized messages to the right people they want to5/7/2016 New York, NY Save This Job
COMPANY OVERVIEW OrthoAccel Technologies, Inc. is a privately owned medical device company engaged in the creation, manufacturing, marketing and sales of innovative solutions that enhance dental care and orthodontic treatment. Among the company?s innovations is AcceleDent, an6/8/2016 Bellaire, TX Save This Job
Marketing Denver, San Francisco or Boston/Burlington, United States Apply Send Jobvite The Solutions Marketing Manager oversees positioning and the go-to-market strategy for Ping Identity?s identity and access management solutions. They are responsible for developing messaging6/14/2016 Denver, CO Save This Job
"Responsible for defining, locating developing, negotiating and closing OEM business relationships within the IT Security, IT Operations and Cloud Service Provider vendor communities." Job Duties: ? Define OEM go-to-market plan, aligned with corporate goals, by researching OEM6/13/2016 Boston, MA Save This Job
We are seeking a hands on Marketing Specialist (Manager, Director, VP level) for the CATV product sector. The ideal candidate will possess a strong background in Product Management and Product Marketing and strategic direction planning with a solid understanding of MSO and data5/1/2016 Carlsbad, CA Save This Job
Location Massachusetts-Corporate - Franklin, MA 2038 US (Primary) Education Bachelor's Degree Category Sales/Marketing Shift Type Standard Business Hours Employee Type Full-time Willingness to Travel 10-25% Interactions is changing the way leading companies communicate with5/10/2016 Franklin, MA Save This Job
Marketing Alameda, California Description Position at 24/7 WEB PROGRAM MANAGER 247 is redefining the way that companies interact with consumers. The company?s customer engagement platform assists several hundred million visitors across all channels, and engages in 1.5 billion5/26/2016 Alameda, CA Save This Job
Marketing ? San Francisco DESCRIPTION Base CRM is an innovative SaaS company with offices in both Mountain View, CA and San Francisco, CA. We are the leading next-generation sales platform that is dramatically improving the way companies are managing their sales and customer5/22/2016 San Francisco, CA Save This Job
Location:Audubon, PA Job Code:DBDIR Description The Director, Business Development and Investor Relations (D-BD) is responsible for all activities related to the review and assessment of new technologies for the company, whether strategically targeted or otherwise, surgeon5/23/2016 Audubon, PA Save This Job
E-Poll Market Research, a dynamic market research company offering product and custom solutions to entertainment and media clients, is seeking a Vice President, Sales. The company has grown and evolved into a leading provider of syndicated products such as E-Score Celebrity and4/26/2016 Encino, LA Save This Job
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Sales Austin , Texas APPLY Responsibilities: Manage existing partners and the growth of new prospective Distribution, Channel, System Integration and Consultants to strengthen market and vertical sales/support coverage Train partners to qualify target accounts and6/13/2016 Austin, TX Save This Job
As an integral part of the sales and marketing team, the Lead Generation Representative responsibilities will include demand creation, qualifying potential customers, increasing awareness of Voxware? product and services, maintaining information in the company?s CRM, and handling5/18/2016 Atlanta, GA Save This Job
Job Description: Solectek Corporation is a leading manufacturer of long-range broadband wireless products used to transmit voice, data and video, including video surveillance and IP video. Founded in 1989 in San Diego, CA (Sorrento Valley), the company is a pioneer in4/30/2016 San Diego, CA Save This Job
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subset: commoncrawl


text:
24 Jun 16
Welcome – 歡迎光臨 – Bienvenue – ようこそ – Welkom – أهلاً وسهلاً Willkommen – स्वागत – Benvenuto – ברוכים הבאים – Bienvenido – Добро пожаловать – Bem-vindo – ยินดีต้อนรับ – välkommen! – καλωσόρισμα
Capital Markets
Georges Ugeux speaks about the reform of the Capital Markets in Iran. After 38 years, he returned to Iran multiple times in 2016 to assess the situation of Iranian Capital Markets and its re-connection to the rest of the world.
View Article (Mona Nezamivand, Bankdari Ayandeh Magazine)
The Flying Dragon Amazon Giveaway ends July 18!
Enter for a chance to win The Flying Dragon (Kindle): An intense, imaginative and intriguing financial thriller. Read More!
13 Jun 16
01 Jun 16
04 May 16
24 Apr 16
Developing a personal website has been an idea of mine for a long time.
The initial purpose was to aggregate my diverse activities and present it in one comprehensive location. I also intend to use it as a platform to promote ideas as well as other people and institutions and to serve as a resource to readers. This project was quite a journey – one that I am pleased to share with you.


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Mature man smiling, close-up, portrait
Credit: Amos Morgan
Creative #: AA044069
License type: Royalty-freeRoyalty-free pricing is based solely on the size of the product you need, not the specific use. You don't have to pay any additional royalties on a use-by-use basis. Once you license a royalty-free product, you may use it multiple times for multiple projects without paying additional fees. (Pornographic, defamatory, libelous or otherwise unlawful use of any image is, of course, prohibited.)
Collection: Photodisc
Release info: Model released
- 50-59 Years,
- Adults Only,
- Business,
- Business Person,
- Businessman,
- Chinese Ethnicity,
- Close-up,
- Color Image,
- Full Suit,
- Gray Hair,
- Headshot,
- Human Body Part,
- Human Face,
- Looking At Camera,
- Mature Adult,
- Necktie,
- One Man Only,
- One Mature Man Only,
- One Person,
- Photography,
- Portrait,
- Studio Shot,
- Suit,
- Toothy Smile,
- Vertical,
- Well-dressed,
- White Background,
Similar imagesSearch ResultsSame series
Mature Man Smiling Closeup Portrait Stock Photo50-59 Years,Adults Only,Business,Business Person,Businessman,Chinese Ethnicity,Close-up,Color Image,Full Suit,Gray Hair,Headshot,Human Body Part,Human Face,Looking At Camera,Mature Adult,Necktie,One Man Only,One Mature Man Only,One Person,Photography,Portrait,Studio Shot,Suit,Toothy Smile,Vertical,Well-dressed,White BackgroundPhotographer Amos MorganCollection: Photodisc


text:
New Delhi, June 29: Security has been tightened at Mumbai airport in view of attack at international airport in Istanbul . Get all the latest... ...http://www.oneindia.com/india/get-latest-news-updates-monday-june-29-here-2139901.html
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DETROIT -- In separate interviews before Tuesday night's All-Star Game, Baltimore second baseman Brian Roberts and closer B.J. Ryan both predicted that their teammate, Miguel Tejada, was destined to stand apart from the crowd. They refrained from anointing him as the Most Valuable Player-in-waiting. But just barely.
Sources: Durant-Thunder meeting 'went well'
6mMarc Stein
Source: Howard seeks a $24M per season deal
18mCalvin Watkins
Day not letting up at no-cut WGC
6mJason Sobel
Hamlin: 'I made a mistake, I didn't let Tony win'
4hBob Pockrass
Michigan State recruit Miles Bridges maxes out vertical test
7hMyron Medcalf
Ranking Tyson's 10 greatest knockouts
7hDan Rafael
1 Related
Collins: Tyson's tumultuous career
9hNigel Collins
Sources: Clips to sell Durant on forming 'Big 4'
4hRamona Shelburne and Arash Markazi
Sources: Heat tout Bosh, Spoelstra to free agents
8hMichael Wallace
Cowboys LB McClain suspended 10 games
4hTodd Archer and Adam Schefter
What to expect from free agency
6hScott Burnside
Anatomy of the Weber-Subban blockbuster
7hPierre LeBrun
Michael Phelps embraces his historic moment
10hWayne Drehs
Photos: A World Cup for the rest of the world
5hPhotography and reporting by Pete Kiehart for ESPN


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Please remember to follow the manual of style and code of conduct at all times.
Check BNN and Bulbanews for up-to-date Pokémon news and discuss it on the forums or in our IRC channel #bulbagarden on irc.systemnet.info.
Check BNN and Bulbanews for up-to-date Pokémon news and discuss it on the forums or in our IRC channel #bulbagarden on irc.systemnet.info.
Kabutops (Fossil 9)
From Bulbapedia, the community-driven Pokémon encyclopedia.
Revision as of 21:24, 24 March 2013 by Maverick Nate (Talk | contribs)
Kabutops (Japanese: カブトプス Kabutops) is a Fighting-type Stage 2 Pokémon card. It is part of the Fossil expansion.
Card text
Pokédex data
|Kabutops - Shellfish Pokémon
|4'3" (1.3 m)
|89 lbs. (40.5 kg)
|Pokédex entry
|Its sleek shape is perfect for swimming. It slashes prey with its claws and drains the body fluids.
Release information
This card was released in the English Fossil expansion as both a Holofoil and Non Holofoil card, originating from the Japanese Mystery of the Fossils as only a Holofoil card. The Non Holofoil print of this card was later reprinted in the English Legendary Collection expansion.
Although this Pokémon is level 30, Kabuto does not evolve into Kabutops until level 40.
Absorb is a move in the Pokémon games that Kabutops can learn. This card's English Pokédex entry comes from Pokémon Red and Blue, whereas the Japanese entry comes from Pokémon Red and Green.
|This article is part of Project TCG, a Bulbapedia project that aims to report on every aspect of the Pokémon Trading Card Game.
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- Seven new Pokémon revealed in new Sun and Moon trailer: Zygarde 50% Forme confirmed to have ...
- Shiny Xerneas codes to be available at Hyper Japan 2016 in London: To be available July 15 to 17


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Horsham Jobs
Looking for jobs in Horsham? Snagajob has great opportunities with the area's leading companies. And applying for Horsham jobs has never been easier. Look at the job listings below, click on a job title that interests you and hit the "apply now" button. Part-time jobs, teen jobs, student jobs and summer jobs are now just a click away. Start your job search now.
Horsham, PA Job Openings
UPSHorsham & Surrounding Areas
Horsham, PA
Qdoba100 Welsh Rd
Horsham, PA
Toys "R" Us/Babies "R" UsHorsham, PA
Horsham, PA
BioReference Laboratories200 Witmer Drive
Horsham, PA
Toys "R" Us/Babies "R" UsHorsham, PA
Horsham, PA
Qdoba100 Welsh Rd
Horsham, PA
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- Doylestown, PA (93 jobs)


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Written on: 17/05/2014 by Tyreez1987 (1 review written)
This company maliciously gained information off an employee and tricked us into an agreement. Possibly looking at losing my business after only being open one month. Extremely sad individuals and disgraceful company. AVOID!!! AVOID !! AVOID!!
Written on: 28/01/2015
We just closing after bein with em 4months worked hard us for ten yrs to be shut like that
As rated by our community of reviewers
" Utility warehouse is fair, reasonable with pricing, deal with queri... " Read More
" Absolute outrage is an understatement. I moved out of there rented ... " Read More
" Problem sorted quickly " Read More
" AVOID AT ALL COSTS " Read More
" NEVER USE THESE PEOPLE " Read More
Written by NPOWERHATER


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Big distributions, little RAM 6
It's that time again where I install the major, full desktop, distributions into a limited hardware machine and report on how they perform. Once again, and like before, I've decided to re-run my previous tests this time using the following distributions:
Fedora 18 (GNOME)
Fedora 18 (KDE)
Fedora 19 (GNOME
Fedora 19 (KDE)
Kubuntu 13.04 (KDE)
Linux Mint 15 (Cinnamon)
Linux Mint 15 (MATE)
Mageia 3 (GNOME)
Mageia 3 (KDE)
OpenSUSE 12.3 (GNOME)
OpenSUSE 12.3 (KDE)
Ubuntu 13.04 (Unity)
Xubuntu 13.04 (Xfce)
I even happened to have a Windows 7 (64-bit) VM lying around and, while I think you would be a fool to run a 64-bit OS on the limited test hardware, I've included as sort of a benchmark.
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SUSE Linux Enterprise 12 SP2 to Ship with GNOME 3.20, Public Beta Out Now
Today, June 30, 2016, SUSE has had the great pleasure of announcing the availabilty of a public beta release of its upcoming, commercial SUSE Linux Enterprise 12 Service Pack 2 operating system.
Review: Linux Mint 18 (Sarah)Portugal vs Poland Live Stream Poland vs Portugal Live Streaming
Review: Linux Mint 18 (Sarah)
If you were looking to jump the Ubuntu ship completely, then we recommend taking a look at our recent Review of Fedora 24. It's equally as good as Mint 18 and equally worthy of your consideration. Between Linux Mint 18 and Fedora 24, we reckon it's exciting times in the Linux world. With the exception and onset of the boring world of vanilla Ubuntu releases, Linux feels reinvigorated and fresh once again. Jump on board, because it can only get better from here.
Security Leftovers


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eSpeak is a compact open source software speech synthesizer for English and other languages, for Linux and Windows.
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House For Sale 28 Trevor Hosken Drive, Wiri, Manukau, New Zealand
Manukau5 bedrooms 2 bathrooms houses manukau
To manukau city shopping centre. Here is a good opportunity for a big family or investment. Please note...
8 photos
Similar properties: houses large bedrooms manukau city19/04/2016
House For Sale 16 Waimarino Place, Weymouth, Manukau, New Zealand Property.co.nz
Manukau3 bedrooms 2 bathrooms houses 3 bedrooms manukau
Is enviable situated on a quiet street in popular weymouth village with stunning views across to the manukau...
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House For Sale 25 Randwick Place, Randwick Park, Manukau, New Zealand
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This wonderfully private and very tastefully decorated home is tucked away in the heart of randwick park. Set on a 644m2 landscaped section this home has 3...
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House For Sale 29 Summermist Drive, Northpark, Manukau, New Zealand
Manukau4 bedrooms 3 bathrooms houses 4 bedrooms manukau
All seasons are beautiful in this immaculate and well maintained family home. Nestled down a driveway, the floor plan provides multiple sun washed living areas...
20 photos8/04/2016
House For Sale 12 Cairnsvale Rise, The Gardens, Manukau, New Zealand
Manukau4 bedrooms 3 bathrooms
If you39re looking for value for money then stop the bus. Beautifully presented 4 bedroom home with awesome views looking over alfriston. Modern and trendy...17/04/2016
House For Sale 35 Walmsley Road, Mangere, Manukau, New Zealand
Manukau5 bedrooms 3 bathrooms houses 5 bedrooms manukau
Heres your chance to secure your future with this premium land blocks bordering sought after mangere bridge. Under the proposed unitary plan the zoning is...
15 photos13/04/2016
House For Sale 7 Valencia Place, Manurewa, Manukau, New Zealand
Manukau4 bedrooms 2 bathrooms
This lovely weatherboard home has seen a family grow over the last 18 years and is now ready for new owners. Situated on a quiet street in a great area this...
12 photos20/04/2016
House For Sale 133 Hillside Road, Papatoetoe, Manukau, New Zealand
Manukau3 bedrooms 1 bathroom
3 bedrooms, 1 bathroom, this hillside road family home offers you charm and extravagance of sensational living and entertaining. The house has abundant...
9 photos20/04/2016
House For Sale 14 Burndale Terrace, Manurewa, Manukau, New Zealand
Manukau3 bedrooms 1 bathroom
Recently refurbished this spacious 3 bedroom 1 bathroom home with separate cabin, lies close to all the local manurewa amenities and ticks all the boxes. This...
9 photos22/04/2016
House For Sale 527 Weymouth Road, Weymouth, Manukau, New Zealand
Manukau2 bedrooms 1 bathroom houses 2 bedrooms manukau
This low maintenance brick tile property has been tastefully refurbished throughout and has so much more than meets the eye. With up to the minute deco a brand...
10 photos24/04/2016
House For Sale 487 Wallace Road, Papatoetoe, Manukau, New Zealand
Manukau2 bedrooms 1 bathroom
Excellent opportunity here with room to add value fantastic location with an easy walk to old papatoetoe town centre and train station. Here is your chance to...
5 photos24/04/2016
House For Sale 122 Franklin Avenue, Papatoetoe, Manukau, New Zealand
Manukau2 bedrooms 1 bathroom
This fantastic 2 bedroom 1 bathroom home suits both investors looking for a fantastic rental or first home buyers. Being close to all the local amenities is...
8 photos22/04/2016
House For Sale 520 Weymouth Road, Manurewa, Manukau, New Zealand
Manukau2 bedrooms 1 bathroom
When location counts, you can39t beat this solid brick and tile beauty for convenience. Located within a 1 minute walk to the train station and manurewa shops...
8 photos24/04/2016
House For Sale 887 Wallace Road, Papatoetoe, Manukau, New Zealand
Manukau2 bedrooms 1 bathroom
Excellent oppourtunity here with room to add value fantastic location with an easy walk to old papatoetoe town centre and transition. Here your chance to make...
5 photos22/04/2016
Cheap As Chips
Currently tenanted at $400pwk is this roomy three bedroom home with a great private section crossleased with glimpses of the manukau. Great existing rental or fantastic first home. Phone to view...4/06/2016
92% of the properties for sale have pictures. On average an advert for Condos Manukau has 8.333 pictures.
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Environmental Enforcement: DeMoulas Super Markets to Pay $33,736 in Chemical Reporting Case
DeMoulas Super Markets Inc. has agreed to pay $33,736 to settle claims by the Environmental Protection Agency (EPA) that it failed to follow federal chemical reporting requirements at a Massachusetts warehouse.
According to a complaint filed last month by the EPA's New England office, the company failed to submit a "material safety data sheet" for hazardous chemical Genetron 22, or a list of chemicals including Genetron 22, held at company's Market Basket perishables distribution warehouse in Andover, Mass.
The data sheet should have been submitted to the state emergency response commission, local emergency planning committee, and the local fire department with jurisdiction over the warehouse. Failure to supply the documentation was in contravention of the federal Emergency Planning and Community Right-to-Know to Act, the EPA said.
The Act requires the owner of a facility to report within three months information about hazardous chemicals present at the facility in amounts that exceed minimum threshold levels. Records indicated DeMoulas acquired the chemical on March 4, 2008.
Genetron 22 is a chemical containing chlorodifluoromethane and is used to service refrigeration systems.
The EPA said that DeMoulas Super Markets Inc. also failed to file required chemical inventory forms on March 1, 2008, for the calendar year 2007 for sulfuric acid, considered by the EPA an "extremely hazardous chemical," and for lead, gasoline, diesel fuel, and R507, all considered "hazardous chemicals."
Sulfuric acid is extremely corrosive and presents significant risks from contact, including lung damage from inhalation of vapors, according to the EPA. Diesel fuel is a flammable liquid and vapor and poses health risks from contact, including skin irritation and lung damage. Lead presents a reactivity risk and a threat to response personnel from contact, including skin and lung contact.
This week the EPA announced that its New England office was cracking down on improper handling and storage of chemicals in the region after a string of cases where chemicals had been mishandled.
Pharmco Products, a Connecticut company that deals in commercial alcohol and chemicals, agreed this week to pay $164,109 to settle claims leveled by the Environmental Protection Agency that it violated sections of the Federal Clean Air Act and the Emergency Planning and Community Right-to-Know Act.
And in February, Southern Maine Specialities – a metal finisher and electroplating company in Maine – and the Massachusetts-based Lamb & Ritchie roofing company agreed to pay over $38,000 and $32,000 respectively for violations of incorrect storage and handling of chemicals.
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Apple Steps Up Targeted e-Mail Campaign to iTunes Users
TMO Reports - Apple Steps Up Targeted e-Mail Campaign to iTunes Users
by , 10:00 AM EDT, May 12th, 2005
Apple Computer has begun to escalate its campaign of sending targeted e-mail to users of its online iTunes Music Store (iTMS), suggesting different types of music and updating users on new albums from previously bought artists.
The marketing ploy has increased in its intensity over the past few weeks, based upon responses from readers of The Mac Observer. Some readers have reported they have been receiving e-mails for over a month now reminding ITMS users of certain albums they have bought in the past and a 'heads up' that the same group has released new music. It appears only recently some users have begun receiving e-mails suggesting different artists based on previous purchases.
"Because you've downloaded music from iTunes in the past, we thought you'd like to know about their new album," one e-mail from Apple suggested to a TMO reader.
The suggestive selling has long been a favorite of Amazon.com and Netflix.com. The process is known as collaborative filtering, where purchase histories are analyzed to determine what else is likely to interest the buyer of a particular product. This approach allows users to navigate from hits that they know they like to more obscure titles they might never have thought of or heard of.
Netflix began the trend by cross referencing a consumers list of favorite movies with other customers to make movie recommendations. Customers then choose movies that have never seen before and Netflix ships the DVD to the customer using its advanced logistics system.
It appears Apple is at least beginning to do very much the same thing as Netflix in leveraging its library of 1.5 million songs to better sell music that might not otherwise be of interest to people who don't often browse or even search its extensive collection.
Some iTMS users have begun to receive suggestive e-mails, recommending they inquire about certain music.
"Netflixs most valuable asset is not their million strong customer marketing list but rather its detailed understanding of their users' tastes and preferences which allows it to make incisive recommendations of related movie titles," said Australian-based media strategist Mike Walsh. "When you consider the huge number of titles already on DVD, and add that to the explosion in content which will become available when the worlds libraries of movie and television content are digitised and placed online a reliable recommendation engine based on a large sample of customer interactions will be a powerful barrier to entry to new players."
"There's going to be a growing need for entertainment-oriented search technology and for personalization and recommendations," said Michael Ramsay, CEO and chairman of TiVo. "If you like this, you're going to like that."
An Apple spokesperson was not immediately available for comment.
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Latest Games
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Run Ronaldo Run
Ronaldo doing what he does best, eating hamburgers and scoring goals!
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It is your job to capture a secret message from the enemy, but watch out, there are a lot of enemies who are defending the one with the message. Try...
Gates vs. Jobs Battle
Fight as Bill Gates or Steve Jobs as you put the other one in their place. And you have light sabers in this star wars style fighting game.
Viper Challenge
Challenge your self against time and collect all star in less time to make a new record. You will get few obstacles on the way... Good Luck!
Berzerk Ball
Berzerk Ball, the expansion pack of the original Homerun game. A brand new look for the game, new weapons, new items and a sexy new character are all...
Snowboard Rush
The most insane Snowboard flash game ever made! Get huge Air and do 5 crazy stunts, while you earn coins and upgrade your gear!
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Play Extreme Trucks Game and drive the huge monster trucks over the rocky terrain, try to get a personal best by scoring a low time.
Ben 10 Savage Pursuit
Search a mutant-infested jungle for pieces of a mysterious device. Play this and other free online games at Cartoon Network.


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Phoenix cops arrest prostitutes, detain them without trial in churches, pressure them to take deals without access to lawyersCory Doctorow
Boing Boing
Mar. 03, 2014
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Putin on Brexit: "Some Don't Want to Dissolve National Borders"
Molly Crabapple sez, "I wrote this piece about a program in Phoenix called Project ROSE arrests sex workers in massive raids and brings them to a church, where they are held extra-judicially and offered alternative sentences without lawyers, judges, or due process."
Monica is a graduate of DIGNITY Diversion. Forced into this program by another prostitution arrest, Monica sat in a classroom from 8 AM to 4 PM, without food, while vice cops described girls overdosing on heroin. Jail was held over the heads of attendees until they finished the program, though many were going broke from their loss of sex-work income. Monica described the class as having the religious overtones of an Alcoholics Anonymous meeting. In keeping with the program's Catholicism, no condoms were provided. Neither was child care.Project ROSE Is Arresting Sex Workers in Arizona to Save Their Souls


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For more than 100 years The Salvation Army has been helping homeless people in Auckland through the Epsom Lodge Supportive Accommodation Unit, giving thousands of people more than just a roof over their heads.
At his lowest point, even hospitals and mental health units refused to see Max as he was too out of control. Today he's a completely different man, helping others through his experiences.
Carla* sold all her furniture and then began stealing to 'feed her kids'. Only her 'kids' were the pokie machines she had developed an addiction to, which she hated, but just could not break.
When I first walked into The Salvation Army I was what you'd call a broken woman
As the controversial TV show Married at First Sight hits our screens, we look at a real life arranged marriage and ask, 'who has got it right—them or us?'
Ken hit rock bottom as an alcoholic truck driver. But Jesus turned that around.
A graduation and a school formal-style celebration in Dunedin at the end of last year encapsulates the work of E&E and the transformation it makes in people's lives.
Helping people find a sense of belonging is at the heart of what Oamaru man Joseph Cropley does. It's led him on a life of teaching and giving back, and to being called a 'hero' by the students whose lives he's changed.
The Salvation Army is strengthening its resourcing for those serving as chaplains.


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*As a Cleveland Browns rookie in 2005, Braylon Edwards announced he'd give $10,000 in scholarships to 100 area eighth-graders if they could graduate high school with at least a 2.5 grade-point average and 15 hours' community service.
Six years later, 79 met the criteria and have begun their first year of college in campuses all across the country, including Harvard, reports the LA Times.
"Without this scholarship, I probably wouldn't be here," Bowling Green freshman David Gholston told ESPN's Rick Reilly.
Now, most people would think writing $790,000 in checks would be enough, but Edwards went one step further. In addition to the $10,000, he provided each of the students with a laptop computer.
"I'm supposed to give people a chance like I was given a chance," Edwards said.
But that's not the only amazing part of the season. Edwards is now a member of the San Francisco 49ers. His salary for this season? $1 million. In other words, he has donated almost his entire salary this season to help those 79 kids.
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August 05, 2014
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Structured Query Language (SQL) Jobs in the East Midlands
Jobs 1 to 15 of 92
.NET Developer, Leicester, 40-45k, plus bens.
I have an excellent opportunity for a .NET Developer to join an industry leading, global company that are growing rapidly... As the .NET Developer you will be working in a team of experts to deliver great results... As the .NET Developer, you must possess the following Requirements: Strong C# Skills...
Leicester, Leicestershire -
Austin Fraser
Salary: £40000 - £45000 per annum + bensPosted: 2 days ago
Senior Developer, Leicester, Up-to 50k plus bens.
I have an excellent opportunity for an experienced Developer to join an industry leading, global company that are growing rapidly. With office's all over the world, this is the right time to join a company that is going from strength to strength. This role offers great career prospects and...
Leicester, Leicestershire -
Austin Fraser
Salary: £45000 - £50000 per annum + BensPosted: 2 days ago
.Net Developer
C# / ASP.NET / SQL / HTML / CSS / JAVASCRIPT... You will utilise your C#, ASP.Net, HTML, CSS, and SQL skills to help us to continue building the best software solutions on the market... C# Net. HTML. CSS. SQL Server. JavaScript... jQuery. Windows Forms... Key Skills: C# / ASP.NET / HTML / CSS / JAVASCRIPT /...
Chesterfield, Derbyshire -
Salary: £30000 - £38000 per annumPosted: 2 days ago
.Net Developer
ASP.Net / C# / SQL Server... You will utilise your ASP.Net, C#, and SQL Server skills to help us stay at the top of the food chain in our sector of the market... C# SQL Server. IT'S NICE TO HAVE… JavaScript. JavaScript. OOP... Key Skills: ASP.NET / C# / MVC /...
Nottingham -
Salary: £30000 - £35000 per annumPosted: 7 days ago
Junior .Net Developer
C# / ASP.NET / SQL / HTML / CSS / JAVASCRIPT... You will utilise your C#, ASP.Net, HTML, CSS, and SQL skills to help us to continue building the best software solutions on the market... C# Net. HTML. CSS. SQL Server. JavaScript... jQuery. Windows Forms... Key Skills: C# / ASP.NET / HTML / CSS / JAVASCRIPT /...
Chesterfield, Derbyshire -
Salary: £20000 - £30000 per annumPosted: Yesterday
VB.Net/C# Developer
Following a year of growth, the time has come to recruit another VB.Net/C# Developer within the team. You will be an established VB.Net Developer with 3 years + experience in VB.Net, C#, SQL and ASP.Net... You will be assisting the team develop IT solutions in VB...
Daventry, Northamptonshire -
I4 Recruitment
Salary: £35k - 44k per year + Benefits+BonusPosted: 11 hours ago
.Net Developer
Net developer required for a company based in North Nottingham .Net Developer Your new company The opportunity have arisen to work for a growing apparel supplier based on the north side of Nottingham... What you'll need to succeed You will need a broad experience of the Microsoft development tools...
Sutton-In-Ashfield -
Hays Specialist Recruitment Ltd
Salary: £25000.00 - £30000.00 per annumPosted: 16 days ago
Developer - Operations Team
Developer (Operations team) Leicester Permanent We're working with a leading retailer in Leicester who are looking for a couple of support developers to join their HR and Finance Systems team due to continued growth over the past 18 months. Responsibilities: - Maintain and improve the infrastructure estate to meet continuously...
Leicester, Leicestershire -
Uniting Ambition
Salary: £22k - 40k per year + bonus, holidays, pension, discounts, childcare vPosted: Yesterday
Graduate .Net Developer - Derby - up to £22k
Visual Studio, C#, ASP.Net, Developer, Development, Software, HTML, CSS, T-SQL, Windows, Web, SQL, IIS, Derby- up to £22k. Looking for an exciting opportunity to join a growing development team? We have a chance for a .Net Developer to join an expanding and financially software company. You will be...
Derby, Derbyshire -
Ascent People Ltd
Salary: £22000 per annumPosted: Yesterday
Senior C# ASP.Net Developer
To be considered for this exciting opportunity candidates must have some commercial experience in the following: *ASP.NET. *C# *MVC. *SQL Server. *Agile experience. The successful Senior C# / ASP.NET Software Developer will have the chance to work on a number of projects with companies from a national to international...
Nottingham, Nottinghamshire -
Senitor Associates
Salary: £30000 - £60000 per annum + Excellent Benefits PackagePosted: 8 hours ago
Graduate / Junior Software Developers x2
Derby, up to £27,000p/a DOE plus benefits, easily commutable from Derby, Nottingham, Leicester and the surrounding areas. Have you recently graduated (including 2016) or you're a self-taught developer and you're looking for your first or next real career opportunity? Do you want to work for...
Derby -
Salary: up to £27,000p/a DOE plus benefitsPosted: 15 days ago
Senior IT Application Developer- Northampton
Senior IT Application Developer (C#/VB.Net/SQL Developer), Northampton... If you are looking for a role where you can: - Use your experience of VB/C# .Net, SQL, ASP.Net/Ajax stack to solve complex problems - Learn a challenging and complex problem domain - Work in small focussed project teams using...
Northampton -
Opus Energy
Salary: £38000.0000 - £45000.0000 per annum + Bonus & BenefitsPosted: 15 days ago
Support & Delivery Engineer - SQL Server, IIS
Support Engineer / Delivery / Application Support / SQL Server / Windows / IIS / SQL / Permanent / Leicestershire / East Midlands / £30,000. Established software house based in Leicestershire are looking for a passionate Support Engineer to join their growing team. The primary purpose of the role is to conduct investigations into client issues and ensure that...
Leicester, Leicestershire -
Spring Technology
Salary: £24000.00 - £30000.00 per annumPosted: 11 hours ago
SQL Server Database Developer - £37K - Leicester
Computer People are proud to be working with a true global market leader in their search for a SQL Database Developer... At the same time you can take advantage of mentoring and training from the Database Development Manager so your career will really flourish!... The primary duties will be involve...
Leicester, Leicestershire -
Computer People
Salary: £30000.00 - £37000.00 per annumPosted: 8 hours ago
Software Developer - C# .Net, - Nottingham - £45k - remote work
A brand new exciting opportunity has arisen for a Software Developer (C#.Net, MVC, SQL Server) to join a highly innovative HR technology solutions company based in Nottingham. As the Software Developer (C#.Net, MVC, SQL Server) you will be joining a dynamic and passionate team of individuals to develop...
Nottingham, Nottinghamshire -
Salary: £40000 - £45000 per annumPosted: 2 days ago
SQL Jobs in the East Midlands


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Favorite films
Recent activityAll
Recent reviewsMore
The Conjuring 2 2016
I'm not mad that I paid to see The Conjuring 2, and that's a victory in and of itself.
Despite a few cringeworthy decisions that I sincerely wish they would have reconsidered (yes, I am absolutely talking about that hilariously horrific CGI Crooked Man), The Conjuring 2 is a decent addition to James Wan's collection of stylish, almost nostalgic horror. His attention to detail is always especially impressive.
And what an oddly beautiful moment it was to be curled up…
They Came Together 2014
Second watch and still a goddamn utter delight. There is not a single thing I don't love about this movie.
Popular reviewsMore
Taken 2008
Whatever, I love this movie. And although I'm sure my own father would try his best to rescue me, I'd like to officially request that Liam Neeson be the one assigned to come after me if I ever get kidnapped and sold into human trafficking. He seems to know his way around an International kidnapping, whereas my dad knows his way around Pawn Stars and getting the best deals on deli meats.


text:
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Something rotten in Ireland's Israeli embassy
international | anti-war / imperialism | other press Tuesday December 18, 2012 16:23 by maolcholann - none Dublin
A response to the latest in a series of provocative, offensive, and frankly fratboyish Facebook posts coming from Israels embassy in Ireland.
Theres Something Rotten in Ireland
Comments (6 of 6)
If Jesus were alive today, both he and Mary would probably be murdered trying to by-pass the ZIo-Nazi Wall around Bethlehemby Zionism=Nazism Wed Dec 19, 2012 13:26
If Jesus were alive today, both he and Mary would probably be murdered by a Zio-Nazi soldier or a Zio-Nazi colonist/"settler".
The message appeared on the IsraelinIreland Facebook page which is linked to on the official embassy site on Monday morning. The post comprised a painting of Mary and Jesus, accompanied by the following caption:
The message sparked immediate heated debate, but was taken down within hours.
Zio-Nazi propaganda reaches new heights of stupidity.
Never underestimate the mendacity, blatant racism and all-round stupidity of your average Zio-Nazi.
They really have no idea how ordinary sane people think.
by Zionism=Nazism Wed Dec 19, 2012 13:33
Banksy's Christmas Card shows Joseph and Mary walled-out of Bethlehem.
by Biblical wise Man. Thu Dec 20, 2012 15:13
Herod was looking for the Gold and Frankincense and Myrr starched away by Mary and Joseph.
by Anto Fri Dec 21, 2012 14:06
by BUTSeriously Thu Dec 27, 2012 01:13
Jerusalem & Hebron ain't no settlements. Jews have never occupied another peoples' land in all their 4000 year history - despite being dispersed throughout the nations. How about you?
Ranting while treating the Bible as an historical source can be detrimental to the little that remains of your sanityby @ The Troll Thu Dec 27, 2012 19:36
Could you show us the evidence you have that "Moses" ever even existed?


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...I've worked with dozens of students who endured Algebra for the first time, and I know what works and what doesn't. I'm a strong authority when it comes to getting my students to understand, in part because of my experience, and in part because I know the curriculum and common test practices very, very well. Algebra 2, in most high schools, follows the year where a student takes Geometry.
Needham, MAView Tutor
...In my many years as a homeroom teacher and advisor, I have gained extensive experience in helping students develop effective study skills and organizational strategies.I have the expertise to tutor elementary students (K-6th). I have 12+ years of experience teaching students in grades 4-6. My sp...
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...Precalculus is both preparation for a following calculus course, AND often a last course in discrete math, so it covers a wide range of topics, some of which are not specifically useful to calculus (such as probability and combinatorics). It relies on solid skills from previous courses in Algeb...
Acton, MAView Tutor
I love to tutor and am excited to find myself with enough time to take on a small number of students. I graduated from MIT and am currently working on a start-up part time and at MIT as an instructor. I miss the one-on-one academic environment and am keen to share some of my knowledge.
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...In that time I have learned a ton, met many inspiring people, and generally had a great time. Every student is different. I like to tailor my specific methods to each individual.
Belmont, MAView Tutor


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Summer Worship Schedule
5:15 pm Worship Service
8:30 am Worship Service
10:00 Worship Service (outdoor, weather permitting)
This Weekend
July 2 - July 3
Seeing the Blessings We Have
We often are more aware of what we think we lack than what we already have. Faith gives us a lens to see all of the amazing blessings God has already given.
Upcoming Events
July 4
Independence Day - Office Closed
You can help us Invite Neighbors to St. John's at the Fourth of July Parade!
We meet at 8:45 am in the Fellowship Hall to
1) hand out FREE cold water bottles to parade goers
2) put flyers in the windshields of cars in our parking lot and
3) bring up the end of the parade with our St. John's banner.
It's fun and will take only an hour.
Kids are welcome!
July 19
Summer Outreach Evening
Newborn Kit Assembly
July 25-29
Vacation Bible School
For information on any of these events, please contact us here
St John's Preschool
June 28, 2016
St John's Lutheran Church
20275Davidson Road
Brookfield, WI 53045
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