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Vaccines targeting glycan structures at the surface of pathogenic microbes must overcome the inherent T cell–independent nature of immune responses against glycans. Carbohydrate conjugate vaccines achieve this by coupling bacterial polysaccharides to a carrier protein that recruits heterologous CD4 T cells to help B cell maturation. Yet they most often produce low- to medium-affinity immune responses of limited duration in immunologically fit individuals and disappointing results in the elderly and immunocompromised patients. Here, we hypothesized that these limitations result from suboptimal T cell help. To produce the next generation of more efficacious conjugate vaccines, we have explored a synthetic design aimed at focusing both B cell and T cell recognition to a single short glycan displayed at the surface of a virus-like particle. We tested and established the proof of concept of this approach for 2 serotypes of
Zinaida Polonskaya, Shenglou Deng, Anita Sarkar, Lisa Kain, Marta Comellas-Aragones, Craig S. McKay, Katarzyna Kaczanowska, Marie Holt, Ryan McBride, Valle Palomo, Kevin M. Self, Seth Taylor, Adriana Irimia, Sanjay R. Mehta, Jennifer M. Dan, Matthew Brigger, Shane Crotty, Stephen P. Schoenberger, James C. Paulson, Ian A. Wilson, Paul B. Savage, M.G. Finn, Luc Teyton
Direct delivery of aerosolized vaccines to the respiratory mucosa elicits both systemic and mucosal responses. This vaccine strategy has not been tested for Ebola virus (EBOV) or other hemorrhagic fever viruses. Here, we examined the immunogenicity and protective efficacy of an aerosolized human parainfluenza virus type 3–vectored vaccine that expresses the glycoprotein (GP) of EBOV (HPIV3/EboGP) delivered to the respiratory tract. Rhesus macaques were vaccinated with aerosolized HPIV3/EboGP, liquid HPIV3/EboGP, or an unrelated, intramuscular, Venezuelan equine encephalitis replicon vaccine expressing EBOV GP. Serum and mucosal samples from aerosolized HPIV3/EboGP recipients exhibited high EBOV-specific IgG, IgA, and neutralizing antibody titers, which exceeded or equaled titers observed in liquid recipients. The HPIV3/EboGP vaccine induced an EBOV-specific cellular response that was greatest in the lungs and yielded polyfunctional CD8+ T cells, including a subset that expressed CD103 (αE integrin), and CD4+ T helper cells that were predominately type 1. The magnitude of the CD4+ T cell response was greater in aerosol vaccinees. The HPIV3/EboGP vaccine produced a more robust cell-mediated and humoral immune response than the systemic replicon vaccine. Moreover, 1 aerosol HPIV3/EboGP dose conferred 100% protection to macaques exposed to EBOV. Aerosol vaccination represents a useful and feasible vaccination mode that can be implemented with ease in a filovirus disease outbreak situation.
Michelle Meyer, Tania Garron, Ndongala M. Lubaki, Chad E. Mire, Karla A. Fenton, Curtis Klages, Gene G. Olinger, Thomas W. Geisbert, Peter L. Collins, Alexander Bukreyev
Recombinant adenoviral vectors (rAds) are lead vaccine candidates for protection against a variety of pathogens, including Ebola, HIV, tuberculosis, and malaria, due to their ability to potently induce T cell immunity in humans. However, the ability to induce protective cellular immunity varies among rAds. Here, we assessed the mechanisms that control the potency of CD8 T cell responses in murine models following vaccination with human-, chimpanzee-, and simian-derived rAds encoding SIV-Gag antigen (Ag). After rAd vaccination, we quantified Ag expression and performed expression profiling of innate immune response genes in the draining lymph node. Human-derived rAd5 and chimpanzee-derived chAd3 were the most potent rAds and induced high and persistent Ag expression with low innate gene activation, while less potent rAds induced less Ag expression and robustly induced innate immunity genes that were primarily associated with IFN signaling. Abrogation of type I IFN or stimulator of IFN genes (STING) signaling increased Ag expression and accelerated CD8 T cell response kinetics but did not alter memory responses or protection. These findings reveal that the magnitude of rAd-induced memory CD8 T cell immune responses correlates with Ag expression but is independent of IFN and STING and provide criteria for optimizing protective CD8 T cell immunity with rAd vaccines.
Kylie M. Quinn, Daniel E. Zak, Andreia Costa, Ayako Yamamoto, Kathrin Kastenmuller, Brenna J. Hill, Geoffrey M. Lynn, Patricia A. Darrah, Ross W.B. Lindsay, Lingshu Wang, Cheng Cheng, Alfredo Nicosia, Antonella Folgori, Stefano Colloca, Riccardo Cortese, Emma Gostick, David A. Price, Jason G.D. Gall, Mario Roederer, Alan Aderem, Robert A. Seder
During seasonal influenza epidemics, disease burden is shouldered predominantly by the very young and the elderly. Elderly individuals are particularly affected, in part because vaccine efficacy wanes with age. This has been linked to a reduced ability to induce a robust serum antibody response. Here, we show that this is due to reduced quantities of vaccine-specific antibodies, rather than a lack of antibody avidity or affinity. We measured levels of vaccine-specific plasmablasts by ELISPOT 1 week after immunization of young and elderly adults with inactivated seasonal influenza vaccine. Plasmablast-derived polyclonal antibodies (PPAbs) were generated from bulk-cultured B cells, while recombinant monoclonal antibodies (re-mAbs) were produced from single plasmablasts. The frequency of vaccine-specific plasmablasts and the concentration of PPAbs were lower in the elderly than in young adults, whereas the yields of secreted IgG per plasmablast were not different. Differences were not detected in the overall vaccine-specific avidity or affinity of PPAbs and re-mAbs between the 2 age groups. In contrast, reactivity of the antibodies induced by the inactivated seasonal influenza vaccine toward the 2009 pandemic H1N1 virus, which was not present in the vaccine, was higher in the elderly than in the young. These results indicate that the inferior antibody response to influenza vaccination in the elderly is primarily due to reduced quantities of vaccine-specific antibodies. They also suggest that exposure history affects the cross-reactivity of vaccination-induced antibodies.
Sanae Sasaki, Meghan Sullivan, Carlos F. Narvaez, Tyson H. Holmes, David Furman, Nai-Ying Zheng, Madhuri Nishtala, Jens Wrammert, Kenneth Smith, Judith A. James, Cornelia L. Dekker, Mark M. Davis, Patrick C. Wilson, Harry B. Greenberg, Xiao-Song He
Diarrheal diseases represent a major health burden in developing countries. Parenteral immunization typically does not induce efficient protection against enteropathogens because it does not stimulate migration of immune cells to the gut. Retinoic acid (RA) is critical for gut immunity, inducing upregulation of gut-homing receptors on activated T cells. In this study, we have demonstrated that RA can redirect immune responses elicited by s.c. vaccination of mice from skin-draining inguinal LNs (ingLNs) to the gut. When present during priming, RA induced robust upregulation of gut-homing receptors in ingLNs, imprinting gut-homing capacity on T cells. Concurrently, RA triggered the generation of gut-tropic IgA+ plasma cells in ingLNs and raised the levels of antigen-specific IgA in the intestinal lumen and blood. RA applied s.c. in vivo induced autonomous RA production in ingLN DCs, further driving efficient induction of gut-homing molecules on effector cells. Importantly, RA-supplemented s.c. immunization elicited a potent immune response in the small intestine that protected mice from cholera toxin–induced diarrhea and diminished bacterial loads in Peyer patches after oral infection with Salmonella. Thus, the use of RA as a gut-homing navigator represents a powerful tool to induce protective immunity in the intestine after s.c. immunization, offering what we believe to be a novel approach for vaccination against enteropathogens.
Swantje I. Hammerschmidt, Michaela Friedrichsen, Jasmin Boelter, Marcin Lyszkiewicz, Elisabeth Kremmer, Oliver Pabst, Reinhold Förster
The in vivo therapeutic efficacy of DC-based cancer vaccines is limited by suboptimal DC maturation protocols. Although delivery of TLR adjuvants systemically boosts DC-based cancer vaccine efficacy, it could also increase toxicity. Here, we have engineered a drug-inducible, composite activation receptor for DCs (referred to herein as DC-CAR) comprising the TLR adaptor MyD88, the CD40 cytoplasmic region, and 2 ligand-binding FKBP12 domains. Administration of a lipid-permeant dimerizing ligand (AP1903) induced oligomerization and activation of this fusion protein, which we termed iMyD88/CD40. AP1903 administration to vaccinated mice enabled prolonged and targeted activation of iMyD88/CD40-modified DCs. Compared with conventionally matured DCs, AP1903-activated iMyD88/CD40-DCs had increased activation of proinflammatory MAPKs. AP1903-activated iMyD88/CD40-transduced human or mouse DCs also produced higher levels of Th1 cytokines, showed improved migration in vivo, and enhanced both antigen-specific CD8+ T cell responses and innate NK cell responses. Furthermore, treatment with AP1903 in vaccinated mice led to robust antitumor immunity against preestablished E.G7-OVA lymphomas and aggressive B16.F10 tumors. Thus, the iMyD88/CD40 unified "switch" effectively and safely replaced exogenous adjuvant cocktails, allowing remote and sustained DC activation in vivo. DC "licensing" through iMyD88/CD40 may represent a mechanism by which to exploit the natural synergy between the TLR and CD40 signaling pathways in DCs using a single small molecule drug and could augment the efficacy of antitumor DC-based vaccines.
Priyadharshini Narayanan, Natalia Lapteva, Mamatha Seethammagari, Jonathan M. Levitt, Kevin M. Slawin, David M. Spencer
Worldwide rates of systemic fungal infections, including three of the major pathogens responsible for such infections in North America (Coccidioides posadasii, Histoplasma capsulatum, and Blastomyces dermatitidis), have soared recently, spurring interest in developing vaccines. The development of Th1 cells is believed to be crucial for protective immunity against pathogenic fungi, whereas the role of Th17 cells is vigorously debated. In models of primary fungal infection, some studies have shown that Th17 cells mediate resistance, while others have shown that they promote disease pathology. Here, we have shown that Th1 immunity is dispensable and that fungus-specific Th17 cells are sufficient for vaccine-induced protection against lethal pulmonary infection with B. dermatitidis in mice. Further, vaccine-induced Th17 cells were necessary and sufficient to protect against the three major systemic mycoses in North America. Mechanistically, Th17 cells engendered protection by recruiting and activating neutrophils and macrophages to the alveolar space, while the induction of Th17 cells and acquisition of vaccine immunity unexpectedly required the adapter molecule Myd88 but not the fungal pathogen recognition receptor Dectin-1. These data suggest that human vaccines against systemic fungal infections should be designed to induce Th17 cells if they are to be effective.
Marcel Wüthrich, Benjamin Gern, Chiung Yu Hung, Karen Ersland, Nicole Rocco, John Pick-Jacobs, Kevin Galles, Hanna Filutowicz, Thomas Warner, Michael Evans, Garry Cole, Bruce Klein
Induction of CD8+ T cell immunity is a key characteristic of an effective vaccine. For safety reasons, human vaccination strategies largely use attenuated nonreplicating or weakly replicating poxvirus-based vectors, but these often elicit poor CD8+ T cell immunity and might not result in optimal protection. Recent studies have suggested that virulence is directly linked to immunogenicity, but the molecular mechanisms underlying optimal CD8+ T cell responses remain to be defined. Here, using natural and recombinant vaccinia virus (VACV) strains, we have shown in mice that VACV strains of differing virulence induce distinct levels of T cell memory because of the differential use of TNF receptor (TNFR) family costimulatory receptors. With strongly replicating (i.e., virulent) VACV, the TNFR family costimulatory receptors OX40 (also known as CD134) and CD27 were engaged and promoted the generation of high numbers of memory CD8+ T cells, which protected against a lethal virus challenge in the absence of other mechanisms, including antibody and help from CD4+ T cells. In contrast, weakly replicating (i.e., low-virulence) VACV strains were poor at eliciting protective CD8+ T cell memory, as only the Ig family costimulatory receptor CD28 was engaged, and not OX40 or CD27. Our results suggest that the virulence of a virus dictates costimulatory receptor usage to determine the level of protective CD8+ T cell immunity.
Shahram Salek-Ardakani, Rachel Flynn, Ramon Arens, Hideo Yagita, Geoffrey L. Smith, Jannie Borst, Stephen P. Schoenberger, Michael Croft
New prophylactic approaches are needed to control infection with the Gram-positive bacterium Staphylococcus aureus, which is a major cause of nosocomial and community-acquired infections. To develop these, greater understanding of protective immunity against S. aureus infection is needed. Human immunity to extracellular Gram-positive bacterial pathogens is primarily mediated by opsonic killing (OPK) via antibodies specific for surface polysaccharides. S. aureus expresses two such antigens, capsular polysaccharide (CP) and poly-N-acetyl glucosamine (PNAG). Here, we have shown that immunization-induced polyclonal animal antisera and monoclonal antibodies specific for either CP or PNAG antigens have excellent in vitro OPK activity in human blood but that when mixed together they show potent interference in OPK activity. In addition, reductions in antibody binding to the bacterial surface, complement deposition, and passive protection were seen in two mouse models of S. aureus infection. Electron microscopy, isothermal calorimetry, and surface plasmon resonance indicated that antibodies to CP and PNAG bound together via an apparent idiotype–anti-idiotype interaction. This interaction was also found in sera from humans with S. aureus bacteremia. These findings suggest that the lack of effective immunity to S. aureus infections in humans could be due, in part, to interference in OPK when antibodies to CP and PNAG antigens are both present. This information could be used to better design S. aureus vaccine components.
David Skurnik, Massimo Merighi, Martha Grout, Mihaela Gadjeva, Tomas Maira-Litran, Maria Ericsson, Donald A. Goldmann, Susan S. Huang, Rupak Datta, Jean C. Lee, Gerald B. Pier
Therapeutic anticancer vaccines are designed to boost patients' immune responses to tumors. One approach is to use a viral vector to deliver antigen to in situ DCs, which then activate tumor-specific T cell and antibody responses. However, vector-specific neutralizing antibodies and suppressive cell populations such as Tregs remain great challenges to the efficacy of this approach. We report here that an alphavirus vector, packaged in virus-like replicon particles (VRP) and capable of efficiently infecting DCs, could be repeatedly administered to patients with metastatic cancer expressing the tumor antigen carcinoembryonic antigen (CEA) and that it overcame high titers of neutralizing antibodies and elevated Treg levels to induce clinically relevant CEA-specific T cell and antibody responses. The CEA-specific antibodies mediated antibody-dependent cellular cytotoxicity against tumor cells from human colorectal cancer metastases. In addition, patients with CEA-specific T cell responses exhibited longer overall survival. These data suggest that VRP-based vectors can overcome the presence of neutralizing antibodies to break tolerance to self antigen and may be clinically useful for immunotherapy in the setting of tumor-induced immunosuppression.
Michael A. Morse, Amy C. Hobeika, Takuya Osada, Peter Berglund, Bolyn Hubby, Sarah Negri, Donna Niedzwiecki, Gayathri R. Devi, Bruce K. Burnett, Timothy M. Clay, Jonathan Smith, H. Kim Lyerly
Development of a vaccine that targets blood-stage malaria parasites is imperative if we are to sustainably reduce the morbidity and mortality caused by this infection. Such a vaccine should elicit long-lasting immune responses against conserved determinants in the parasite population. Most blood-stage vaccines, however, induce protective antibodies against surface antigens, which tend to be polymorphic. Cell-mediated responses, on the other hand, offer the theoretical advantage of targeting internal antigens that are more likely to be conserved. Nonetheless, few of the current blood-stage vaccine candidates are able to harness vigorous T cell immunity. Here, we present what we believe to be a novel blood-stage whole-organism vaccine that, by combining low doses of killed parasite with CpG-oligodeoxynucleotide (CpG-ODN) adjuvant, was able to elicit strong and cross-reactive T cell responses in mice. Our data demonstrate that immunization of mice with 1,000 killed parasites in CpG-ODN engendered durable and cross-strain protection by inducing a vigorous response that was dependent on CD4+ T cells, IFN-γ, and nitric oxide. If applicable to humans, this approach should facilitate the generation of robust, cross-reactive T cell responses against malaria as well as antigen availability for vaccine manufacture.
Alberto Pinzon-Charry, Virginia McPhun, Vivian Kienzle, Chakrit Hirunpetcharat, Christian Engwerda, James McCarthy, Michael F. Good
Vaccinia virus (VV) vaccination is used to immunize against smallpox and historically was considered to have been successful if a skin lesion formed at the vaccination site. While antibody responses have been widely proposed as a correlate of efficacy and protection in humans, the role of cellular and humoral immunity in VV-associated skin lesion formation was unknown. We therefore investigated whether long-term residual humoral and cellular immune memory to VV, persisting 30 years after vaccination, could control VV-induced skin lesion in revaccinated individuals. Here, we have shown that residual VV-specific IFN-γ+TNF-α+ or IFN-γ+IL-2+ CD4+ lymphocytes but not CD8+ effector/memory lymphocytes expressing a skin-homing marker are inversely associated with the size of the skin lesion formed in response to revaccination. Indeed, high numbers of residual effector T cells were associated with lower VV skin lesion size after revaccination. In contrast, long-term residual VV-specific neutralizing antibody (NAbs) titers did not affect skin lesion formation. However, the size of the skin lesion strongly correlated with high levels of NAbs boosted after revaccination. These findings demonstrate a potential role for VV-specific CD4+ responses at the site of VV-associated skin lesion, thereby providing new insight into immune responses at these sites and potentially contributing to the development of new approaches to measure the efficacy of VV vaccination.
Bénédicte Puissant-Lubrano, Philippe Bossi, Frederick Gay, Jean-Marc Crance, Olivia Bonduelle, Daniel Garin, François Bricaire, Brigitte Autran, Behazine Combadière
Vaccine strategies that utilize human DCs to enhance antitumor immunity have yet to realize their full potential. Approaches that optimally target a spectrum of antigens to DCs are urgently needed. Here we report the development of a platform for loading DCs with antigen. It is based on killed but metabolically active (KBMA) recombinant Listeria monocytogenes and facilitates both antigen delivery and maturation of human DCs. Highly attenuated KBMA L. monocytogenes were engineered to express an epitope of the melanoma-associated antigen MelanA/Mart-1 that is recognized by human CD8+ T cells when presented by the MHC class I molecule HLA-A*0201. The engineered KBMA L. monocytogenes induced human DC upregulation of costimulatory molecules and secretion of pro-Th1 cytokines and type I interferons, leading to effective priming of Mart-1–specific human CD8+ T cells and lysis of patient-derived melanoma cells. KBMA L. monocytogenes expressing full-length NY-ESO-1 protein, another melanoma-associated antigen, delivered the antigen for presentation by MHC class I and class II molecules independent of the MHC haplotype of the DC donor. A mouse therapeutic tumor model was used to show that KBMA L. monocytogenes efficiently targeted APCs in vivo to induce protective antitumor responses. Together, our data demonstrate that KBMA L. monocytogenes may be a powerful platform that can both deliver recombinant antigen to DCs for presentation and provide a potent DC-maturation stimulus, making it a potential cancer vaccine candidate.
Mojca Skoberne, Alice Yewdall, Keith S. Bahjat, Emmanuelle Godefroy, Peter Lauer, Edward Lemmens, Weiqun Liu, Will Luckett, Meredith Leong, Thomas W. Dubensky, Dirk G. Brockstedt, Nina Bhardwaj
Recombinant adeno-associated virus (rAAV) vectors were used in human trials as carriers of vaccines for HIV-1 after encouraging preclinical results. However, the clinical trials yielded disappointing results. Here we demonstrated that in mice, rAAV vectors expressing the gene encoding HIV-1 gag stimulated gag-specific CD8+ T cells, but these T cells failed to expand after a booster immunization with a replication-defective adenoviral (Ad) vector also expressing gag. We tested rAAV vectors of different serotypes expressing HIV-1 gag for induction of transgene product–specific CD8+ T cells and found that the immunoinhibitory effect of rAAV priming observed with different AAV serotypes was transgene product specific, was independent of the interval between prime and boost, and extended to boosts with vaccine modalities other than Ad vectors. rAAV vector–induced CD8+ T cells proliferated poorly, produced low levels of IFN-γ in response to gag stimulation, and upregulated immunoinhibitory molecules. These T cells did not protect efficiently against challenge with a surrogate pathogen. Finally, we showed that the impaired proliferative capacity of the T cells was caused by persistence of the antigen-encoding rAAV vectors and could be reversed by placing the CD8+ T cells in an antigen-free environment. Our data suggest that rAAV vectors induce functionally impaired T cells and could dampen the immune response to a natural infection.
Shih-Wen Lin, Scott E. Hensley, Nia Tatsis, Marcio O. Lasaro, Hildegund C.J. Ertl
Initial immune responses to allergens may occur before birth, thereby modulating the subsequent development of atopy. This paradigm remains controversial, however, due to the inability to identify antigen-specific T cells in cord blood. The advent of MHC tetramers has revolutionized the detection of antigen-specific T cells. Tetramer staining of cord blood after CMV infection has demonstrated that effective CD8+ antigen-specific immune responses can follow intrauterine viral infections. We hypothesized that sensitization to antigens occurs in utero in humans. We studied cord blood B and T cell immune responses following vaccination against influenza during pregnancy. Anti-Fluzone and anti-matrix protein IgM antibodies were detected in 38.5% (27 of 70) and 40.0% (28 of 70), respectively, of cord blood specimens. Using MHC tetramers, HA-specific CD4+ T cells were detected among 25.0% (3 of 12) and 42.9% (6 of 14) of cord blood specimens possessing DRB1*0101 and DRB1*0401 HLA types, respectively, and were detected even when the DRB1 HLA type was inherited from the father. Matrix protein–specific CD8+ T cells were detected among 10.0% (2 of 20) of HLA-A*0201+ newborns. These results suggest that B and T cell immune responses occur in the fetus following vaccination against influenza and have important implications for determining when immune responses to environmental exposures begin.
Deepa Rastogi, Chaodong Wang, Xia Mao, Cynthia Lendor, Paul B. Rothman, Rachel L. Miller
Bacterial vectors may offer many advantages over other antigen delivery systems for cancer vaccines. We engineered a Salmonella typhimuriumvaccine strain to deliver the NY-ESO-1 tumor antigen (S. typhimurium–NY-ESO-1) through a type III protein secretion system. The S. typhimurium–NY-ESO-1 construct elicited NY-ESO-1–specific CD8+ and CD4+ T cells from peripheral blood lymphocytes ofcancer patients in vitro. Oral administration of S. typhimurium–NY-ESO-1 to mice resulted in the regression of established NY-ESO-1–expressing tumors. Intratumoral inoculation of S. typhimurium–NY-ESO-1 to NY-ESO-1–negative tumors resulted in delivery of antigen in vivo and led to tumor regression in the presence of preexisting NY-ESO-1–specific CD8+ T cells. Specific T cell responses against at least 2 unrelated tumor antigens not contained in the vaccine were observed, demonstrating epitope spreading. We propose that antigen delivery through the S. typhimuriumtype III secretion system is a promising novel strategy for cancer vaccine development.
Hiroyoshi Nishikawa, Eiichi Sato, Gabriel Briones, Li-Mei Chen, Mitsutoshi Matsuo, Yasuhiro Nagata, Gerd Ritter, Elke Jäger, Hideki Nomura, Shigeto Kondo, Isao Tawara, Takuma Kato, Hiroshi Shiku, Lloyd J. Old, Jorge E. Galán, Sacha Gnjatic
Immunotherapy for the treatment of metastatic melanoma remains a major clinical challenge. The melanoma microenvironment may lead to local T cell tolerance in part through downregulation of costimulatory molecules, such as B7.1 (CD80). We report the results from the first clinical trial, to our knowledge, using a recombinant vaccinia virus expressing B7.1 (rV-B7.1) for monthly intralesional vaccination of accessible melanoma lesions. A standard 2-dose–escalation phase I clinical trial was conducted with 12 patients. The approach was well tolerated with only low-grade fever, myalgias, and fatigue reported and 2 patients experiencing vitiligo. An objective partial response was observed in 1 patient and disease stabilization in 2 patients, 1 of whom is alive without disease 59 months following vaccination. All patients demonstrated an increase in postvaccination antibody and T cell responses against vaccinia virus. Systemic immunity was tested in HLA-A*0201 patients who demonstrated an increased frequency of gp100 and T cells specific to melanoma antigen recognized by T cells 1 (MART-1), also known as Melan-A, by ELISPOT assay following local rV-B7.1 vaccination. Local immunity was evaluated by quantitative real-time RT-PCR, which suggested that tumor regression was associated with increased expression of CD8 and IFN-γ. The local delivery of vaccinia virus expressing B7.1 was well tolerated and represents an innovative strategy for altering the local tumor microenvironment in patients with melanoma.
Howard L. Kaufman, Gail DeRaffele, Josephine Mitcham, Dorota Moroziewicz, Seth M. Cohen, Karl S. Hurst-Wicker, Ken Cheung, David S. Lee, Joseph Divito, Magalese Voulo, Julie Donovan, Kate Dolan, Kelledy Manson, Dennis Panicali, Ena Wang, Heidi Hörig, Francesco M. Marincola
Current models of T cell memory implicate a critical role for IL-7 in the effector-to-memory transition, raising the possibility that IL-7 therapy might enhance vaccine responses. IL-7 has not been studied, to our knowledge, before now for adjuvant activity. We administered recombinant human IL-7 (rhIL-7) to mice during immunization against the male antigen HY and compared these results with those obtained from mice immunized with rhIL-2 and rhIL-15. Administration of rhIL-7 or rhIL-15, but not rhIL-2, increased effector cells directed against these dominant antigens and dramatically enhanced CD8+ effectors to subdominant antigens. The mechanisms by which the cytokines augmented effector pool generation were multifactorial and included rhIL-7–mediated costimulation and rhIL-15–mediated augmentation of the proliferative burst. The contraction phase of the antigen-specific response was exaggerated in cytokine-treated mice; however, CD8+ memory pools in rhIL-7– or rhIL-15–treated groups demonstrated superior long-term survival resulting in quantitative advantages that remained long after the cytokines were discontinued, as demonstrated by improved survival after challenge with an HY-expressing tumor undertaken several weeks after cytokine cessation. These results confirm the adjuvant activity of rhIL-15 and demonstrate that rhIL-7 also serves as a potent vaccine adjuvant that broadens immunity by augmenting responses to subdominant antigens and improving the survival of the CD8+ T cell memory pool.
Fraia Melchionda, Terry J. Fry, Matthew J. Milliron, Melissa A. McKirdy, Yutaka Tagaya, Crystal L. Mackall
Pseudomonas aeruginosa is an important opportunistic pathogen that can cause chronic and often life-threatening infections of the respiratory tract, particularly in individuals with cystic fibrosis (CF). Because infections with P. aeruginosa remain the major cause of the high morbidity and mortality of CF, a vaccine against P. aeruginosa would be very useful for preventing this disorder. The outer membrane protein F (OprF) of P. aeruginosa is a promising vaccine candidate and various B cell epitopes within OprF have been identified. Given that adenovirus (Ad) vectors have strong immunogenic potential and can function as adjuvants for genetic vaccines, the present study evaluates the immunogenic and protective properties of a novel replication-deficient Ad vector in which the Ad hexon protein was modified to include a 14–amino acid epitope of P. aeruginosa OprF (Epi8) in loop 1 of the hypervariable region 5 of the hexon (AdZ.Epi8). Immunization of C57BL/6 mice with AdZ.Epi8 resulted in detectable serum anti–P. aeruginosa and anti-OprF humoral responses. These responses were haplotype dependent, with higher serum anti-OprF titers in CBA mice than in BALB/c or C57BL/6 mice. AdZ.Epi8 induced Epi8-specific IFN-γ–positive CD4 and CD8 T cell responses and resulted in protection against a lethal pulmonary challenge with agar-encapsulated P. aeruginosa. Importantly, repeated administration of AdZ.Epi8 resulted in boosting of the anti-OprF humoral and anti-Epi8 cellular response, whereas no boosting effect was present in the response against the transgene β-galactosidase. These observations suggest that Ad vectors expressing pathogen epitopes in their capsid will protect against an extracellular pathogen and will allow boosting of the epitope-specific humoral response with repeated administration, a strategy that should prove useful in developing Ad vectors as vaccines where humoral immunity will be protective.
Stefan Worgall, Anja Krause, Michael Rivara, Kyung-Kim Hee, Enrico V. Vintayen, Neil R. Hackett, Peter W. Roelvink, Joseph T. Bruder, Thomas J. Wickham, Imre Kovesdi, Ronald G. Crystal
DCs are critical for priming adaptive immune responses to foreign antigens. However, the utility of harnessing these cells in vivo to optimize the immunogenicity of vaccines has not been fully explored. Here we investigate a novel vaccine approach that involves delivering synergistic signals that both recruit and expand DC populations at the site of antigen production. Intramuscular injection of an unadjuvanted HIV-1 envelope (env) DNA vaccine recruited few DCs to the injection site and elicited low-frequency, env-specific immune responses in mice. Coadministration of plasmids encoding the chemokine macrophage inflammatory protein-1α (MIP-1α) and the DC-specific growth factor fms-like tyrosine kinase 3 ligand with the DNA vaccine resulted in the recruitment, expansion, and activation of large numbers of DCs at the site of inoculation. Consistent with these findings, coadministration of these plasmid cytokines also markedly augmented DNA vaccine--–elicited cellular and humoral immune responses and increased protective efficacy against challenge with recombinant vaccinia virus. These data suggest that the availability of mature DCs at the site of inoculation is a critical rate-limiting factor for DNA vaccine immunogenicity. Synergistic recruitment and expansion of DCs in vivo may prove a practical strategy for overcoming this limitation and potentiating immune responses to vaccines as well as other immunotherapeutic strategies.
Shawn M. Sumida, Paul F. McKay, Diana M. Truitt, Michael G. Kishko, Janelle C. Arthur, Michael S. Seaman, Shawn S. Jackson, Darci A. Gorgone, Michelle A. Lifton, Norman L. Letvin, Dan H. Barouch
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What Makes The ARs so Accurate?
The first thing that makes ARs so accurate is triggers. Without a good trigger no rifle can reach its potential. The first ARs and indeed, some of the later ones, had atrocious triggers. Full of creep, poundage and travel! All the things you dont want in any trigger, be it in a bolt action or the semi auto AR. Because of shooting games using ARs, and (former) bolt rifle shooters accustomed to fine triggers, theres been a push for better equipment. Several manufacturers have met that need in their own and differing ways. Makers like Geissele, Timney, Jewell, American Gold Trigger, and Rock River all make excellent AR triggers that make the rifles a lot more accurate and enjoyable to shoot. I personally love the Rock Rivers two stage match trigger and the American Gold. They are, for all intents and purposes, bench rest triggers for semi autos. To install them, you simply pop out two pins, remove the old trigger and stick the replacement back in. Its that easy and they make a world of difference in shooting comfort and accuracy!
This is a picture of The Beast, my AR10 Matt Salm built for me. Good looking rig, right? It has a 26 barrel and a .875 muzzle behind the Vais brake. It should shoot an 85 grain pill all the way to 600 yards to bring grief to PDs! (Did you notice the Scope Shield Alaska scope cover dangling from it? Or the awesome Night Force 8x32 scope?) Oops, I have left off one of my valuable pieces of equipment, one I dont shoot without! Its the 3BUCC shell catcher. That is a required item if you want to keep brass to reload or simply not to litter. Works great!
I finally got some quality range time. Two groups like this and one more. All three were less than .600 and this five shot group went .425 with the four into .179. Not bad for an old guy with a gas gun. If only that one...
Another important component I believe that is critical to fine accuracy is a good scope mounted properly in reliable rings. One must remember that a scope on an AR gets recoil in two directions. I dont believe you can mount a cheap scope in good rings nor a good scope in cheap rings and expect it to perform as well as a good scope in properly secured good rings. Read that again! If youre looking for the utmost accuracy and repeatability out of your AR, you simply cannot cut corners. I didnt get the groups pictured in this article without attention to the details of properly mounting a quality scope in quality rings. My favorite rings are TPS. Ive never had to lap any of them and they come with important but very simple directions for mounting. Nothing moves in these mounts if you follow their directions.
The third key component is the barrel and chambering. Some would argue it is the first component, but I will say all three parts are about equal. You can have them heavy or light, long or as short as you like. Im in the camp of long, cut rifled barrels. Im not a fan of button rifled barrels; however, the factory built ones I have are button rifled and they shoot as accurate as my cut rifled barrels. But, if Im ordering a barrel, it will be cut rifled. There are several small, quality barrel makers and theres Kreiger. My 6x47 Lapua has a Brux barrel and the 20 TAC and the 223A.I. have LRB barrels. Their bores are as smooth as butter, take very little to clean up and shoot fantastically. Kreiger is probably the finest barrel maker today and theirs are cut rifled. I have five of theirs on bolt rifles. In addition, Krieger sets very high standards for the blanks before they accept them and then even higher standards for their finished work. You cant go wrong with Krieger nor the other two companies, Brux and LRB.
These three components are, in my humble opinion, the most important parts of what will make an AR accurate, or not. All the other parts are simply what are needed for completing the rifle and the look the builder and owner are striving for. However, if the builder cuts any corners in the build, even the best components wont matter. He still has to do all of his work properly. The ARs I own I fully enjoy shooting, and they will shoot a lot better than Im capable of doing. They are lots of fun, very accurate, and interesting in their diverse configurations and calibers. Yes, the ARs are the Modern Sporting Rifle. If you havent shot one, what are you waiting for?
I have been blessed by God to have a wonderful life: a life of travel and the opportunity to meet many people who have became good friends. We live in a country filled with diverse beauty and incredibly interesting people. My advice is to get out there and explore our beautiful country and meet new friends. Travel by ground roads, not air, if at all possible! Enjoy the scenic beauty of the lands we live in, the places you travel through and the loveliness of your destination. Go with an open, uncluttered mind that will allow you to see and appreciate it. You will be richer for it. God made it for us to enjoy; I hope you will as I have. Remember too, that getting old is normal but growing up is optional! Try new things, and if you havent tried the modern sporting rifle, what are you waiting for? I encourage you to do so. So many calibers, so many configurations, so many choices. Aint life grand?
My thanks and appreciation to:
Jessica, my sweet niece who proofread this and gave moral support. I couldnt have done it without you!
Phillip, my son, who also aided me in making the corrections that Jessica pointed out.
Lee Mosher of Insight Shooting Systems, Pueblo,Colo. Lee got me started and has remained a great friend. He builds very fine AR uppers!
3BUCC. Builder of the fine brass catchers for ARs. An indispensable item!
Scope Shield Alaska. Probably the finest scope cover made!
Matt Salm, a Patriot. One who built an excellent AR 10 upper.
Don Lahr, Precision Ballistics, LLC, for those fine 6mm bullets.
Bill Alexander, Alexander Arms and his staff for their support and for building a great little AR.
and Rock River and DPMS for building some of the worlds best ARs!
Charles E. Speck is a Virginia Christian clay and target shooter, hunter and reloader. He has been doing this for over forty years and says he is still learning! Charles has written a couple articles and had an article written about a gun he had commissioned called "The Patriot," which is the only gun ever to commemorate 9/11. Charles loves side by sides and accurate rifles.
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text:
Kitchen Garden: What to Grow and How to Grow It
Here's what I love about Kitchen Garden: the charts. Turn to any vegetable and learn when to plant each variety and when to harvest it. Yes, this information is all available in the HEAVY Western Garden's book, but it's so clearly presented in Kitchen Garden that I find myself more inspired when using it than when getting carpal tunnel flipping through WG. Maybe it's a classic case of eliminating overchoice: select information clearly presented sometimes works better than a compendium of everything under the sun.
This totally works! Aphids be gone!Tags: vegetables, tomato, Eco, aluminum, green, gardening, water, hoses, green livingAdded 8 Years Ago from Thebugblaster
An easy way to have fresh herbs within reach to add to salads, sauces and other dishes. The leak proof bags make it so you can grow them straight in the package.
The only way to grow tomatoes, upside-down!Tags: gifts, outdoor, technology, cooking, vegetables, tomato, gourmet, bags, patioAdded 11 Years Ago from Thegreenhead


text:
From: Spain
Genre: Alternative/Alt. Rock, Metal, Grunge
Born in Madrid 1977. Loving music since he was a child and learning by himself... Styles: Rock, Grunge, Punk and Heavy Metal.
His bands were: KNOWLEDGE IS A WEAPON (1999-2003) for 3 years playing bass guitar and one more as vocalist making eclectic Hardcore Metal, SLEAZE (2000-2001) screaming in a Hardcore Punk band, DARTH VADER'S REVENGE (1997) playing bass guitar & MADTV (1995) at the guitar. Now on this own project.
His tracks recorded by himself are available on CD or digital inside ReverbNation…


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[Date Index]
[Thread Index]
[Author Index]
WITH - A web interface to Mathematica
(note, this email address is valid and will be for a few months at
leas. Make the
obvious changes in a few months).
I've written a short open-source PHP script that when installed on a
UNIX/Linux web server with Mathematica, allows use of Mathematica via
any browser. There's support for 2D & 3D graphics, but not the live
interactive graphics.
There's some screen shots on both desktop computers as well as a
handheld PDA (hp iPAQ) at
There are no special requirements for the browser - no need for
cookies, JavaScript etc.
1) This allows access to almost all Mathematica functions, including
Run[] which allows arbitrary commands to be executed.
2) Most (if not possibly all) Mathematica licenses don't allow
unrestricted access,
then WITM must obviously only be installed on a password protected
system, to restrict access. (Apache provides a mechanism for this).
I've not tried it over a Secure Socket Layer (i.e https://...)
connection, but it should work fine on that.
More info on the Souceforge site:
It's licensed under the GNU General Public License.
David Kirkby


text:
Failed Asylum Seekers See Driving Rules Tightened
The Department of Transport says that only people granted permission to stay in the country for 185 days or more will be eligible to take the driving test. This change applies to people from outside the European Economic Area and does not affect those from within it.
The transport minister, Paul Clark, said that it was the right policy. The rule change has been implemented under existing laws, meaning that it will not need to be approved by Parliament or debated by MP's.
In a statement that was written to the House of Commons, Mr Clark said that, as a preliminary measure, he was informing the house that from yesterday on, those who are applying for a provisional, as well as full driving license, will have to demonstrate that they are lawful residents of the UK. They can no longer simply be lawfully present in order to qualify for a driving license.
He went on to say that those who are present in the UK on temporary permission or temporary release under the Emigration Act of 1971 will not be considered eligible fro a driving license. The people granted leave to enter or remain in the UK for at least 185 days will continue to be able to apply for a license whilst their leave is extant – of course, that is provided that they otherwise qualify for such a license. It is right that those who have undecided status should not be able to seek benefits of ordinary life in the UK.
Related Stories:
- Australian activists halt asylum seeker deportation - February 3, 2015
- Canadian government debating stringent new rules for citizenship - May 15, 2014
- London tech companies suffering from strict immigration rules - April 24, 2014
- New Zealand government rows over easing immigration rules - April 17, 2014
- Controversial changes to Norway immigration rules - March 3, 2014
- Canadian citizenship rules to be reformed during 2014 - January 24, 2014
- Archbishop condemns UK immigration rules for separating British families - December 17, 2013
- Battle to save asylum seeker baby and his family from deportation - November 28, 2013
- New Zealand issues new rules for international students - October 11, 2013
- New UK family immigration rules may deter executive relocation - June 12, 2013
Latest News:
- UAE-Indian eMigrate inititative will end exploitation - September 8, 2015
- Egypt seizes vessels with more than 200 migrants - September 7, 2015
- Indian employers angered by eMigrate system - August 18, 2015
- Church leader: African leaders must address problems that spark emigration - August 14, 2015
- 60 migrants missing off Libya coastline - August 13, 2015
- More illegal immigrants found in lorry in UK - August 11, 2015
- Nine illegal migrants seen exiting truck at Essex shopping centre - August 7, 2015
- PNG government promises Manus alcohol crackdown - August 6, 2015
- British landlords told to evict illegal immigrants - August 5, 2015
- Polish family emigrate to Plymouth after seeing images on Google Street View - July 28, 2015


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nicotine stained vertical blinds
Hello! :yes:
I am new to this forum.
I am just wondering how to get the nicotine stain off of white vertical blinds that have been stained yellow from a smoker smoking in the same room as the blinds.
Thank you!
Good luck, I hope you get an answer.:whistling2:
Tried bleach soaking in the bath tub or use the garden hose?
@Tizzer thanks I will try that!
Mr Clean eraser sponges......typically I don't like those things but was quite amazed how well they worked for this very problem.
I tried your suggestion and it seemed to work really well. I could not completely tell because I think the blinds were actually not white originally but beige because I compared them to some other blinds in a room where there was no smoking and they looked almost the same after using the eraser.
Sounds like they were alabaster, more common than white. Glad it worked for you,
Every smoker needing another reason to quit--at least indoors and around other people who may or may not themselves---should do a paint prep of a smoked in space. All that gunk is in the fabrics people sit on and wear. And frankly in their lungs. I cannot think of a better example of second hand smoke. After one job, with an entire family (including the baby of the teen--I guessed) having smoked continuously, I had nightmares of this orange/brown gunked up cleaning fluid dripping on me at night.
For complete paint prep of lots of houses and especially those of smokers? I got in the habit of calling ServiceMaster or other local cleaning companies with an army of people that would do walls, floors and ceilings spotless and ready for primer and paint. I asked for the appropriate fire damage team when doing a smoker house. It was cheaper than clients paying me to do the prep.
Still had to prime it with horrible smelling but effective primers and treat raw rood in basement with enzymes to get rid of the leftover odor before painting.
Anyhow, sounds like the Mr. Clean things worked for the blinds. I have had reasonable success getting discoloration out of blinds soaking in oxy clean. It worked well on vinyl window screens too although I usually turned to Purple Clean or something from the auto parts store to carve the orange tar and gunk loose. Or I would pressure wash at low pressure with an industrial cleaner.
This is in response to your inquiry back in 07 about cleaning verticle blinds: if they aren't fabric covered, try 'Scrubbing Bubbles', the bath cleaner. I used this product on vinyl horizontal mini-blinds. I removed them, draped over my deck railing, fully extended, then sprayed with Scrubbing Bubbles. Immediately the foaming bath cleanser started doing 'it's stuff'! They became pristine white before your eyes! It even cleaned the nylon cords, etc! I used a garden hose to wash them off and allowed air dry outside then re-hung. They looked just like they did when new!
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NFL Draft > 2010 Prospects > OG > Sergio Render
Sergio Render Scouting Report
|Virginia Tech Hokies
|Jersey Number:
|Height & Weight:
|6'3 - 319 lbs.
2010 NFL Draft Prospect Scouting Report:
Sergio Render, OG, Virginia Tech
Off-season shoulder surgery has not slowed down Sergio Render. Ever since he stepped foot on Virginia Tech's campus, Render has been starting at one of the offensive guard spots. During his freshman campaign he earned the starting right guard job and he has never given it up, although he has moved to the left guard spot for his senior season. At 6-3 and 319 pounds, Render is a big guard who has turned into a great leader on the line. The Hokies had some questions about their ground game heading into the 2009 season after projected starter Darren Evans went down with an ACL injury. All the Hokies had to turn to were inexperienced players. Redshirt freshman Ryan Williams is easily the Atlantic Coast Conference's top rusher six weeks into the 2009 season and Williams has Render to largely thank for that success. Heading into the 2009 season Render was considered by most as the nation's top offensive guard. His size and versatility are a big bonus and he should be off the board by the second round -- barring a major injury. Render could even play his way into the first round -- which is tough for any guard -- if he keeps going at this pace. 1/17 Update: Render had a good senior season, but most scouts have yet to pay him much attention. Render had a superb bowl game against Tennessee, but for some reason he is still not garnering too much national attention. At this point it looks like Render will have to make a name for himself during the workouts leading up to the draft in order to be considered a second or even third round option.
Last Updated Jan-17-2010 by Joel Welser
0 Comments | Add Yours
There are no comments on this prospect yet.


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Feb. 3, 2014 – TaxTalk Blog
Are you excited to receive your tax refund this year? Unfortunately, tax fraud criminals are also looking forward to stealing your refund. If you think this won't happen to you, know that identity theft criminals stole $24.7 billion from the nation in 2012, according to the U.S. Department of Justice.
Identity theft is a growing epidemic that is especially prominent during tax season. This is the time when taxpayers gather all of their most sensitive information into one place, making it easiest for criminals to steal information and tax returns.
But don't worry. This year, the department has implemented an identity protection program to help all Hoosiers protect themselves during the 2014 tax season.
The identity protection program will offer resources for taxpayers to help them guard against their personal information being stolen.
The most crucial aspect of protecting your identity is protecting your Social Security number (SSN) and card. Don't give out your SSN unless you have to, and always ask why it's needed, how it's going to be used and how it will be stored. What is most important to remember is that you should never give your SSN to a person with whom you do not make initial contact. For example, if someone calls or emails you, do not give them your SSN.
It is safe to give your SSN to government agencies including the Indiana Department of Revenue and Internal Revenue Service (IRS) and to your employer.
Additionally, be aware of the most common ways thieves can access your information. Here are some of the ways identity theft criminals can find your personal information:
The department has also partnered with the Office of the Indiana Attorney General to help taxpayers be aware of ways to best protect themselves from the devastating and increasing crime of identity theft. The Attorney General's Office is offering an online ID Theft Protection Toolkit, which acts as a central hub for taxpayers to check their credit, make sure their identities haven't been compromised, alert credit companies when necessary and freeze their credit if their identities have been compromised.
Throughout the week, the department will continue to deliver TaxTalk blogs with information about how you can protect your identity. Stay tuned for Wednesday's post for tips on filing and storing your documents safely.
If you would like to submit a question or topic suggestions, please send them to firstname.lastname@example.com
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Full Archive...


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|Thread Tools
I have a very lightly used Marantz VP15S1 projector. We used this in our showroom as a demo unit. I need to find out the exact hours on it but it can't be more 1000, maybe less. Can anyone give me an idea of what this is currently worth? Thank you.
Not near as much as it should:
My Baffle wall LCR build: http://www.avsforum.com/forum/155-di...-tpl-150h.html
I have been watching the VP15s for a few months and almost bought the one that sold for $1600 but went with the Planar PD8150 from an AVS member. I would say the 2 are in the same class selling around $2000 if in perfect functional condition and more than 1/2 lamp life left.
Please note (you probably know this but others looking may not) that the Marantz projectors have a count down timer that starts at 2000 and goes to 0, so if it says 800 there are 1200 hours on the lamp and 800 left.
Good luck with the sale!
Planar PD8150 with DC4 DMD paired with a Dalite HP 2.8 110"
Thank you.
I received the new remote control today from Marantz. The bulb has a WHOPPING total of 119 hours of use. It says there is a lamp life left of 1885 hrs. I knew it was basically new as we only used for short demos. I can tell you for sure this projector is the most babied projector ever. Professional a/v showroom, climate controlled, low low usage, and not a scratch of any kind. With this in mind what is a realistic value for my unit? Any honest guidance would be appreciated.
Mines' worth over $2 easy peasy. Good luck to you too!
Login to Ebay, enter Marantz VP15, click advanced search options, click sold,click search.. $1500 is the most recent going price.. and will drop as time goes by. Do you have a projection setup in your home? If not why not use it?
Planar PD8150 with DC4 DMD paired with a Dalite HP 2.8 110"
I already have a home setup or would use it for sure. It's such a nice unit, only 100 hrs. of use. As for ebay I have seen what is for sale, really not the same thing. I guess as far as a model number it is, but far more use than mine. It will sell...............it may take some time but it will sell. Thanks for the reply.
Mark, don't you think that's a bit ingenuous?
Yesterday's MSRP's and reviews (at least many parts of them) are irrelevant in the current market.
If you don't believe that, I'll sell you can have the 486 PC for which I paid $2800 for only $500.
Why is that. He said he would never get more than $1,800 for it. On Ebay in the last six months some sold above and some below. All of the ones I looked at had a lot more hours on them. Mark said to make it a nice package deal with installation. Don't see where that post said to gouge someone. The 15S1 Marantz projector is a 1080P projector, which is the current standard. Your PC is not anywhere close to the current standard. The Marantz throws an image that is as good as many of todays DLP projectors. It just does not have the light output of today's DLP projectors.
My Baffle wall LCR build: http://www.avsforum.com/forum/155-di...-tpl-150h.html
Thanks Mike. Another thumbs up for you. Gave you one too Noah.
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text:
"I absolutely LOVED this card. It was my first time ordering from Tiny Prints - the order was delivered on time as stated and the results were beautiful! I used this as my son's graduation announcement. The only issue I had, and it was a very small issue, was the choice of text size on the initial at the top. I wanted to use something different at the top, but could not because it locks withing a certain font size range. The cards look AWESOME with the pearl paper - it is well worth the extra cost. I can't say enough about how pleased I was!"
"Fantastic graphics. Trifold product is cool and unique. Paper quality feels "rich." When your 18-year-old graduate thinks it's amazing, that says a lot!"
"Loved the pciture quality, the paper, and the entire process was very professional and prompt. Would definately order from this company again."
"The only thing I didn't consider until they came in was that I wanted to write something on the back for each person I sent them to and I had to write in metallic permanent marker for it to be read. I should have thought about that and wonder if there was a choice to be able to have a writing surface on the back. Other than that I loved the finished product!"
"Everyone commented on my son's graduation announcement. It was just great! I loved the larger sizes available too. The shipment arrived days before we expected it and that was a bonus too."
Copyright © 2004-2017 Shutterfly, Inc | Tiny Prints | Terms of Use. Designated trademarks and brands are the property of their respective owners.


text:
Shelby GT350 for Sale
Selected Filters:Type of Ad: Vehicles for Sale Make: Shelby Model: GT350 Remove All
1968 Shelby GT350
St. Louis, MO
Coming soon!... More
Advertiser since 1987
2011 Shelby GT350
St Charles, MO
This 2011 Shelby GT350 for sale has5.0 Liter Supercharged V8 (525 HP), 6 Speed Manual Transmission, 1 of Only 157 Production Vehicles Ever Made,... More
Advertiser since 1993
1969 Shelby GT350 GT 350
Fairhaven, MA
1969 Shelby GT 350 This project GT 350 was running and driving before being taken apart for a restoration. It's a factory pastel grey black interior... More
Advertiser since 2012
1968 Shelby GT350 Convertible
Greenwich, CT
1968 Shelby GT350 Convertible Numbers Matching Accompanied by Deluxe Marti Report 1 of 404 GT 350 Convertibles Built in 1968 Built January 3rd, 1968, this Shelby GT-350 was... More
2015 Shelby GT350 Mustang Coupe
Lake Park, FL
2015 Ford Mustang Shelby GT350!Deep Impact Blue Metallic with Ebony (Black) Cloth and Suede Seats Interior!Only 44 Miles!SHOWROOM CONDITION!! -Loaded!-Bluetooth!-Track Package!-Extremely Rare!-Very LOW Mileage!-Raised Deck... More
2015 Shelby GT350 Mustang Coupe
Lake Park, FL
***COMING SOON*** SHADOW BLACK WITH EBONY LEATHER/SUEDE SEATS! ONLY 43 DELIVERY MILES!! EXTREMELY LOW MILEAGE! NEVER DEALER PREPPED! STRIPE DELETE!! TECHNOLOGY PACKAGE! HEATED/COOLED FRONT... More
1969 Shelby GT350 GT 350
Fairhaven, MA
1969 Shelby GT 350 Candy Apple Red with very nice white interior. This is a factory automatic with it's original 351 engine and AC. This GT 350... More
Advertiser since 2012
2011 Shelby GT350 R
Calgary, AB
2011 Shelby GT350 "R" Supercharged! Extremely rare sought after car, now a collector item due to it being an original Shelby GT350R (even more... More
1970 Shelby GT350 Coupe
Orlando, FL
Very original and showing 53,000 miles. 351 ci V8 engine, automatic transmission, power steering, power brakes, tinted glass, traction lok rear axle, sport deck... More
1967 Shelby GT350 Coupe
Sarasota, FL
REAL documented with full historical documentation. including Shelby Registry Marti report buck tag window sticker including letter from Dave Mathews 1 of 7 total... More
2015 Shelby GT350 Coupe
Lake Park, FL
2015 Ford Mustang Shelby GT350!Race Red with Ebony (Black) Leather and Suede Seats Interior!Only 89 Miles!SHOWROOM CONDITION!! -Loaded!-Extremely Rare!-Very LOW Mileage!-Technology Package!-Heated/Cooled Seats!-Dual Zone Electric... More
2015 Shelby GT350 Coupe
Lake Park, FL
2015 Ford Mustang Shelby GT350!Deep Impact Blue Metallic with Ebony (Black) Leather and Suede Seats Interior!Only 40 Miles!SHOWROOM CONDITION!! -Loaded!-Extremely Rare!-Very LOW Mileage!-Technology Package!-Heated/Cooled Seats!-Dual... More
1968 Shelby GT350 Convertible
St. Louis, MO
To view more pictures of this vehicle, please CLICK HERE! One of only 404 Shelby GT350 Convertibles ever built! One of only 82 in Sunlit... More
Advertiser since 1989
1968 Shelby GT350 Convertible
Farmingdale, NY
1968 Shelby GT350 for sale. Motorcar Classics is proud to present this awesome 1968 Shelby GT350 Convertible featured in Wimbledon White with Black interior... More
1968 Shelby GT350 Convertible with 4-Speed!
Houston, TX
Carlyle Motors is very proud to offer this gorgeous 1968 Shelby GT 350 Convertible with 4-Speed transmission. Gorgeous Wimbledon White paint and Black Vinyl... More


text:
Polo Ralph Lauren has announced that it will further extend its e-commerce reach with the launch of Ralphlauren.com in the UK. The news follows hot on the heels of another top label relenting to the internet's pull.
After the success of Ralph Lauren's US e-boutique, launched in 2000, the company has decided to roll out a British version on October 14. The site will offer a full assortment of clothing and accessories for men, women and children, in addition to a full array of home products. For women that includes the glamorous Ralph Lauren Collection, Black Label, Blue Label and the technical sports apparel line RLX. For men, that includes the refined Purple Label, Black Label and more casual Polo line.
The announcement comes less than two weeks after avant-garde label Maison Martin Margiela joined the world of e-commerce. The e-boutique, launched on October 1, is powered by the fashion retailing site YOOX and is rendered in MS-DOS font, giving it the cool look of a syntax error message. The site sells key pieces from Lignes 0, 10, 22, 11, and 6, as well as shoes, jewelry, menswear, and online exclusive pieces like the reinterpretation of the brand's signature "5 Zips" leather biker jacket.
Just four years ago, 68 percent of top luxury brands did not sell their products online, according to the Retail Bulletin. But wealthy customers are of growing importance to e-commerce. Affluent customers are arguably doing a disproportionate amount of online shopping. Households with incomes of $100K or more comprised 37 percent of total online spending in the second quarter of 2010 while households with incomes of less than $50K comprised 22 percent, according to an eMarketer report.
"Affluent consumers now expect luxury retailers' websites to replicate the same shopping experience they offer in their stores," says Jeffrey Grau, a senior analyst at the firm. "Yet many luxury brands have not risen to the occasion. Several, like Chanel, have been slow to launch fully transactional websites and need to take e-commerce more seriously or will risk alienating their customer base."
Ralph Lauren UK: http://www.ralphlauren.co.uk/home/index.jsp?ab=int_fd_unitedkingdom
Maison Martin Margiela: http://eboutique.maisonmartinmargiela.com/home.asp?tskay=2B0A884FReuse content


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Wasteland 2 - Interview with Chris Keenan
GamingBolt has an interview with Development Director Chris Keenan of inXile Entertainment about Wasteland 2, and various other topics.
Ravi Sinha: With Obsidian Entertainment, Fallout 1 & 2 composer Mark Morgan, Planescape: Torment writer/designer Colin McComb and of course, the producer of the first game, Wasteland 2 is certainly brimming with industry and RPG legends. How was such a team assembled for the project?
Chris Keenan: Everything about the way we've planned, financed and built this project is different from the way we've worked over the past 10-15 years. We have our core implementation team that has worked together for a while and just thought, "If we can have anyone we want for this title, how would we do it?".
Brian immediately reached out to many members from the original team for design help. Being that we are not working with a publisher it, we could make decisions that would have been sticky to figure out, like bringing on Chris Avellone. Being that he's CCO of Obsidian, it would potentially scare publishers due to him knowing "trade secrets" about the game, but Brian has worked with the Obsidian guys for many years and knows he can trust them. We've developed a very close relationship with Obsidian and will continue to scratch each others backs.
Ravi Sinha: We've seen this post-apocalyptic set-up culminate in either finding a MacGuffin to save the world and usher in a new age of man or in exploring the so-called wasteland and making one's own choices a la Fallout. Will Wasteland be in either direction, a mix of both or completely out of left field with its plot?
Chris Keenan: Well, being that Wasteland was the original Fallout, Brian and his team decided to keep a moderately similar high-level story feel. Neither were about saving all of humanity and bringing pixie-dust and smiles to all. Wasteland was all about the moments you came across while trying to bring about a bit of order and navigate issues as they came up.
The setting is pretty bleak and there really is no way to "save the world" even if you wanted to. The citizens of the Wasteland have literally had a trial by fire and after a hundred years of being in pure survival mode, they don't necessarily operate on logic that we'd hold true in our current world.
Ravi Sinha: In regards to its design, was there always that desire to make Wasteland 2 a throwback to the classic RPGs of yore?
Chris Keenan: We went through a bunch of design ideas when thinking about what Wasteland 2 would be, but many of the elements that stuck kept that familiar feel from games of the past. As we continued to communicate the vision to our community prior to the Kickstarter release, we kept hearing how much people missed that classic play experience and knew it was the right decision for the game.
At the time, I think there was a feeling over the game development community that many game systems evolved out of necessity to a more mass market friendly approach. Publishers weren't funding deals unless your game could sell a million units and that generally tends to remove the option of more hardcore game systems from the designs. Our approach is that we don't care about the mass market. Our 65,000+ backers want a more deep detail and stat oriented game.
Information aboutWasteland 2
SP/MP: Single-player
Setting: Post-Apoc
Genre: RPG
Platform: PC
Release: Released


text:
Jun 04
football manager 2014
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The Football Manager 2014 ps vita gaming series has been around for over 20 years now. Starting out as Championship Manager in the early 1990's, Football Manager is widely accepted as the only football management game you need to own. Over the years, many other software houses have tried to dethrone Football Manager but nobody is yet to come remotely close. There is often criticism of developers who release annual updates to their titles that they are simply


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Samoa may have a penchant for the 15-a-side game but they did their best to avoid a rugby-like score as the Olyroos clambered to a 6-0 win at Marconi Stadium last night.
The Australian under-23 side was unconvincing in the opening match of its Olympic qualifying series, taking until the second half to shut out a team playing with 10 men from the 30th minute.
Predictions of a scoreline to match some of those from the 2001 World Cup qualifiers - in which the Socceroos bagged 66 goals in four matches - were unfounded, with the home side clearly short on combination.
"The finishing I thought was poor, some of the crossing was pretty poor," Olyroos coach Frank Farina said.
"I think we were too slack in the first half, and, to give them a bit of credit, they were defending pretty well from some of the crosses that were coming in.
"The second half was a lot better - obviously playing against 10 men was a big advantage."
The Olyroos missed several opportunities in the first half before laying on five goals in 29 minutes during the second.
Parramatta Power midfielder Ahmad Elrich scored a double and Marconi star Alex Brosque looked good in patches but, with just three internationals in three years, the Olyroos looked rusty.
Several of early chances were cruelled by sloppy finishing from the Olyroos before Samoan defender Ted Losi Sikovi downed Luke Wilkshire in the area. The Bristol City midfielder dusted himself off and made it 1-0 from the spot.
But if the Australians thought the penalty would signal the end of the Islanders' resistance, they were in for quite a shock, as it only seemed to stiffen their resolve.
Even the 30th-minute sending off of Sikovi for an apparent elbow to the head of striker Brosque failed to perturb the Samoans as they held out an impotent Olyroo attack until the break.
They were even a touch unlucky not to have levelled the scores after 40 minutes when Michael Toleafoa's chip from 25 metres hit the crossbar.
In the night's earlier match, Solomon Islands beat Papua New Guinea 4-2.
Auckland: New Zealand opened their group B qualifying tournament with a 9-0 win over the Cook Islands.
Tonga beat American Samoa 3-0 in the early game.
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It is traditional that hardware gets more powerful and adds features with every generation - unless you're Sony...
Following on from controversial decisions such as to strip the PS2 hardware emulation from first generation PS3s and the option to install Linux as an alternative OS from the PS3 Slim, Sony has decided to even things up a little: by stripping this latter option from all PlayStation 3s.
The update - which launches on 1 April, though Sony insists isn't an April Fools - will come with firmware v3.21.
"{It} will disable the "Install Other OS" feature that was available on the PS3 systems prior to the current slimmer models, launched in September 2009," said Sony Senior Director of Corporate Communications & Social Media, Patrick Seybold. "This feature enabled users to install an operating system, but due to security concerns, Sony Computer Entertainment will remove the functionality through the 3.21 system software update. In addition, disabling the "Other OS" feature will help ensure that PS3 owners will continue to have access to the broad range of gaming and entertainment content from SCE and its content partners on a more secure system."
Naturally enough, committed Linux users can choose to skip the upgrade, but Sony warns this will mean you lose the following (fundamental) features:
- Ability to sign in to PlayStation Network and use network features that require signing in to PlayStation Network, such as online features of PS3 games and chat
- Playback of PS3 software titles or Blu-ray Disc videos that require PS3 system software version 3.21 or later
- Playback of copyright-protected videos that are stored on a media server (when DTCP-IP is enabled under Settings)
- Use of new features and improvements that are available on PS3 system software 3.21 or later
So understandable elimination of a novelty feature of another prime example of Sony misunderstanding its user community? You let us know...


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Klitschko-Haye Update!
Former world cruiserweight champion David 'The Hayemaker' Haye has offered to box in Germany on a K2 Promotions show in order to revive negotiations for a summer showdown with WBO and IBF world heavyweight champion Wladimir Klitschko. Originally scheduled to take place on June 20 at Stamford Bridge in London, Haye's eagerly anticipated duel with Klitschko has hit a series of snags in recent weeks, and now appears on the brink of collapse. Haye's trainer and manager Adam Booth says the Klitschko camp has distorted some of the facts. "David will fight Wladimir anywhere they choose, and give them the two rematch options they have requested….what they're also demanding is that if David were to win against Wladimir and then have a mandatory challenger obligation (non-Klitschko) he would have to immediately vacate the title he won from Wladimir. This would mean David could win three fights against the Klitschko brothers and still not be in a position to officially call himself heavyweight champion of the world at the end of it. They (the Klitschkos) would make exactly the same money in Germany as they would in England because we have dropped our figure to go to Germany. If they take a proposed fight with Chris Arreola, they will make substantially less. So people can now work this out for themselves."


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Re: Linux always opening new front-end
Happens with KDE3.5 and Enlightenment, too. Agreed: it's an annoyance.
On Wednesday 04 February 2009 03:20:23 Sebastian Meznaric wrote:
> This is not really a bug per se but it is very annoying that if one
> opens a file from GNOME file browser then for each new opened file a
> new front-end pops up. Is there a way to tell Mathematica to check if
> there is already a running front-end and/or kernel before opening a
> new one?
Curtis Osterhoudt
cfo at remove_this.lanl.and_this.gov
PGP Key ID: 0x4DCA2A10
Please avoid sending me Word or PowerPoint attachments
See http://www.gnu.org/philosophy/no-word-attachments.html


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Drama / Characters: 3 male, 4 female
Scenery: Exterior with set pieces and projections
This free structured look at the 1986 Challenger disaster places the teacher who died with six others as they hurtled into space at the center of an exploration of our need to reach beyond ourselves and dare the universe. Defying Gravity artfully interweaves the past with the present and the lives of participants and bystanders, drawing parallels among painter Claude Monet's artistic quest, the zest of the teacher selected to the first civilian astronaut, the perspectives of her grieving daughter, the aspirations of elderly tourists who drive their Winnebago to Florida to watch the space shot and dream of hotels in space, the guilt felt by a NASA mechanic and his girl friend's fear of heights.
"Flies high in its attempt to describe man's fascination with space and its conquest.... You will certainly not be bored." N.Y. Post.
" A] clever and uplifting fantasy ... with] ear catching musings about art, religion and the outer limits of human possibility." N.Y. Times.
"A lovely piece.... It floats gracefully in the big blue yonder of the imagination ... letting Anderson's delicate, tender and human attitude toward her characters come through.... One by one they rise out of their earthbound selves ... to look down on the world from a new perspective." N.Y. Daily News.
|Defying Gravity
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Recasting air force apparel in sweat fabric, these comfortable black sweatpants feature functional thigh pockets and 3D construction that follows the legs more precisely.
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Overwhelming, the majority of solar energy systems in Texas are grid-tied solar… that's actually true the world over. So what does grid-tied solar mean? Simply put, the electricity from the solar energy system is tied in to the buildings existing service panel, supplementing the energy supplied from your utility company.
While grid-tied solar systems use advanced technology, the principles behind how they work are simple. Solar panels convert light energy from the sun into power, which then flows to an inverter where the direct current (DC) power is converted to the alternating current (AC) used in the home. From there, the electricity flows to a breaker(s) in the electric service panel on your home (see system diagram below for more detail) – from which you then consume just as you do from your utility company.
Should your home require more electricity than what is being generated by the grid-tied solar, additional electricity is simply pulled from the local utility company's power grid. On the flip side, excess electricity generated flows back into the grid – actually spinning your electric meter backwards. To make sure you're earning the maximum benefit from your solar array, contact your electricity provider… Green Mountain Power is usually the best choice in Texas.
The grid-tied solar system is THE choice for homeowners looking to lower electric bills, hedge against future rate increases, and all the while lessening your impact on the environment. The main confusion surrounding grid-tied solar is the perception that the system will power the home in the event of an outage – unfortunately it does not, and when we explain why, it'll make perfect sense.
Fortunately, Texas Solar Outfitters offers grid-tied solar systems with battery backup that resolves this issue. To learn more about our battery backup systems, click here.


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What's New Wednesday: Just Swatched
Hi BeautyTalkers! For today's What's New Wednesday, I wanted to show you 3 sets that I just swatched (like, just rubbed the colors off of my arms) for an upcoming email. I can't show you the swatches yet, but the palettes are newly live on the site, so check 'em out.
And believe me when I say, they're beeeeyoutiful in person.
What new sets would you like to see swatched? Show & Tell!
Buxom Shimmer & Shake™ Show-Stopping Collection of 15 Mini Full-On™ Lip Polishes & Creams
bareMinerals Degrees Of Dazzling
Obsessive Compulsive Cosmetics Lip Tar All-Star Mini x 12 Set
The Marc Jacobs lip trove, please and thank you!
I would love to see the lip tars swatched! <3
I'm so thankful that there are smaller lip tars available. Those regular sized tubes may seem small but there's a TON of product in them! So something smaller is much appreciated.
Yes, I would love to see Marc Jacobs Lip Trove. The formula of these lipsticks is wonderful. I own a few and would LOVE to be pushed over the edge to buy this set!!
I really want those lip tars! I can't wait to see what the swatches look like.
@janinebt I agree! Smaller lip tars are perfect! I have several regular sized lip tars and i'm pretty sure I have not use even 10% of it, maybe even 5%! Plus they look so cute now that they made them smaller!
That Buxom set has been in my loves list since I set eyes on it. Thinking I will take out the colors that I love and divide up the others into stockings or something.
Total want that Buxom set big time!!! Love me some Buxom. Kiki I love your idea of gifting the ones you don't like as much.
@shapeshifter & garnett - We have swatched the colors that are in the lip trove! You can see them here: http://community.sephora.com/t5/The-Swatch-Board/Marc-Jacobs-Beauty/m-p/663367 (scroll to the bottom).
Buxom, please! I really want that set, I'm waiting for gc time and then purchasing it along with some Fresh goodies, come on December! lol
I've had a HUGE lip fetish lately & have been eyeing the new Lip Tar Set! I'm with Janine on the regular sizes being too much, even if its primarily for my makeup kit. My sister tried out another one of our lip sets recently, but found it a little drying, but since Buxom is one of her favs, I may encourage her to pick one up
@miabt - I think you mean enable.
I'd love to see swatches of this!
I feel like it's such an interesting set but I'm hesitant to buy without seeing swatches.
^^^ I agree with marydiva, I would love to see this swatched!!! It's been on my Loves List since it popped up online .
@marydiva , i had to try out the irridescent lip gloss in the store, it looks seriously cool swatched on my hand! Unfortunately it looked pretty much clear on my lips. I did think the shadow duo was nice, but not as nice as the one in the MJ set i'm hoping to get.
You must be a registered user to add a comment. If you've already registered, sign in. Otherwise, register and sign in.


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Wednesday, September 8, 2010
27 comments:
- cofc ultimate said...
Try and come to College of Charleston on that college tour...Music farm is a sick place for concerts and its 70 to 30 girl guy ratio no brainer.
- September 8, 2010 at 1:58 PM
- cofc ultimate said...
- This comment has been removed by the author.
- September 8, 2010 at 1:58 PM
- Eyesofphases said...
Very dope
- September 8, 2010 at 2:20 PM
- Ayimme said...
Please say your gonna start selling those
- September 8, 2010 at 4:38 PM
- Sonya said...
Shits sick!!!!!!!!!
- September 8, 2010 at 5:30 PM
- foxxy said...
what ayimme said. you GOTTA sell them shits, man.
- September 8, 2010 at 5:44 PM
- marlee (eclectiq.MIND*) said...
that looks flyyy ! Btw, when are you selling those dreamville shirts ?? On ustream you said about $5 ya know with the recession && all lol . Girl sizes please :)
- September 8, 2010 at 8:56 PM
- Maji said...
hi jcole! that jacket is hot. but anyways, i'm just here to say that I love you so much! and I support you all the way. and i was sooo pissed when i found out you were in Atlanta a few days ago because I missed you. but i love you regardless and i'm so excited for everything that you are doing. and i just get really happy when i hear your songs on the radio because it's so nice to know that other people are showing you love as well. i love you!
- September 8, 2010 at 10:32 PM
- Elite said...
damn homie.. that shit fire... save 1 for me lol
- September 9, 2010 at 12:04 AM
- Mimi said...
Dopeness! i fux w/ Dreamville and all things related.
- September 9, 2010 at 7:16 AM
- piasco said...
that is TOO tough.
- September 10, 2010 at 7:46 AM
- B Mo said...
Quality. Ur truly breathing life back into the Game.
- September 10, 2010 at 8:29 AM
- Le'Shae said...
I just wanted to say that I really like and appreciate your music. I have high hopes for you, but seems like every time I feel that way for a rapper that hasn't made it mainstream yet, they finally make it and turn out to disappoint me because they've watered their music down. So just please continue to do you and don't change for ANYBODY nor ANYTHING!
- September 12, 2010 at 1:32 AM
- Cruziiie said...
WHere is your UNDRCRWN?
Let's get you in one of their varsity jackets....especially with the whole sports lifestyle/basketball vibe the both of you have!
- September 12, 2010 at 12:12 PM
- kenneth said...
Nice! {SupremeFabulosity.wordpress.com} has been trying to get in contact with whoever we need to, so we can interview you. We have interviewed plenty of celebs so far, and are waiting to add your talent to the roster! Thanks! We are on Twitter @Kjuelznycmiami
- September 14, 2010 at 9:42 AM
- Twitty10 said...
Maneee You Gon Wear That Every Got Damn Night?? You No What I Paid For This Shit Bitch You Got Damn Right!!!
- September 16, 2010 at 5:54 PM
- FoxyTheDiva said...
LOve it!
- September 17, 2010 at 5:31 PM
- Joycelynn said...
i want one!!!!
- September 25, 2010 at 12:23 AM
- Will said...
Im tryna get on hit me up email@example.com
- September 25, 2010 at 8:06 PM
- Will said...
Im tryna get on hit me up firstname.lastname@example.com
- September 25, 2010 at 8:07 PM
- Jackie Choice said...
Love it... gotta put on! 2-6 <3
- September 28, 2010 at 8:34 PM
- CheezyDoDo said...
Cole slide me one of those jackets bruh, once the hoes know im on th dreamvillie basketball team i will finnally getting all the groupies i dreamed of, I need this in my life
- September 29, 2010 at 12:07 AM
- lil said...
how do i get one of dem coats
- April 7, 2011 at 12:42 PM
- RecKless NUPE said...
Ths jacket is fly everyone go check out my blog http://recklessreg.blogspot.com/ i just did a great piece on j cole
- August 9, 2011 at 8:45 AM
- coleworld said...
j cole hell's kitchen http://www.youtube.com/watch?v=zSHvHoc6_dU
- October 21, 2013 at 5:17 AM
- coleworld said...
j cole hell's kitchen http://www.youtube.com/watch?v=zSHvHoc6_dU
- October 21, 2013 at 5:17 AM
- coleworld said...
j cole hell's kitchen http://www.youtube.com/watch?v=zSHvHoc6_dU
- October 21, 2013 at 5:18 AM


text: On friday my Tivo S3 got a software update to 11.3b7-01-2-652 and now all my channels except for the basic channels now say "This channel is not authorized". Anyone else having this issue? I called Verizon FIOS because I thought it was their issue, they came out and change the cable cards, cables, ONT and everything looked good on their end. I can even take out the cable cards from the 2-series 3's that are not working and put them in to my Series 4 and I get all the channels. Tivo support said maybe 1-2 weeks for a fix.


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Hello, I'm new to the forums as well as owning a bike for the first time. I have recently bought a Suzuki Intruder 1400. She's a great looking bike and ran well for the first 40 or so miles then I started having clutch problems. First I added new brake fluid, and bled the lines a bit and that worked fine. Then I had the same problem after a week or so. I have ordered a rebuild kit so I can rebuild the clutch. I took the clutch apart and it seems like the new parts are not the same as the old ones. I know that after market parts are usually not the same as the OEM but I wanted some opinions on the subject. Several websites have told me that the parts I ordered are what the bike calls for but I dont want to ruin anything. Any help would be greatly appreciated.
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The Paducah Sun's Easter Egg Hunt Contest went down to the wire Sunday with Virginia Walker of Golconda, Ill., finding the elusive golden egg after the eighth and final clue was published in Sunday's newspaper.
Walker, assisted by daughter Darlene Gibbs, found the egg early Sunday morning near a cedar tree close to the air park near the McCracken County Animal Shelter. Sunday's final clue read: "Don't be late to reach the gate, halfway to the cedar tree is where I'll be." Saturday's seventh clue stated: "Being on the hunt can be helter skelter, but when a storm is nearby seek shelter."
Walker has participated in The Sun's egg hunt the past eight years.
"I just went by the clues," Walker said of her egg-finding strategy. She admitted the first couple of clues did not help her find the prize egg, but by the time she read the final clues and had an idea of the egg's location, she told her daughter, "Now is the time to go check."
When the two found the golden egg there weren't a lot of other egg-hunters in the vicinity, Walker said. As to who saw the egg first, Walker or her daughter, she said, "I guess you could say I did." Walker admits she doesn't get around too well by herself and she was glad to have her daughter accompany her.
Walker won a $500 gift certificate from Hutson Inc., the contest sponsor. And because she is a Paducah Sun subscriber, she also won $500 cash. As for plans for her winnings, "I'll have to think about that ... I'll probably find a good use for it," she said.
According to Kendra Payne Mitchell, marketing manager for The Paducah Sun, there was a concerted effort to make this year's contest a little more challenging. "This is the longest the contest has gone before (a winner claimed the prize)," Mitchell said. "We've never published all eight clues before."
The contest generates a lot of interest, and is promoted on Facebook as well as in the newspaper, Mitchell said. "We really got a lot of positive feedback with people thanking us for making it tougher."
Check out these recently discussed stories and voice your opinion...


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In Newtown, a gun debate does not rage on—at least in public
NEWTOWN, Conn.—As families of the victims of the Sandy Hook Elementary School shootings travel to Hartford to join President Barack Obama at his speech on gun violence later on Monday—and fly with him back to Washington—many residents here said they did not want the gun-control spotlight shone on them.
More than a dozen people—outside the Newtown General Store, Starbucks and Panera Bread, and at Treadwell Memorial Park less than a mile up the hill from Sandy Hook Elementary—declined to be interviewed on the subject. Some said they wanted to keep their opinions to themselves. Others said they felt the media had exploited them enough and wanted to be left alone.
"I thought you all had left," one man said cheerily as he strolled along a walking path in Sandy Hook center on one of the first truly warm days of spring.
Outside Edmond Town Hall, a group of mothers sipped coffee as they watched their children play underneath a sign for a second-run showing of the movie "Argo." They politely declined to speak about guns.
Others were not so cordial. A woman who spotted a reporter's media badge on Church Hill Road as she drove by rolled down her window to inform him, "You should be ashamed."
Ian Eller, a writer and father of two Sandy Hook students who wrote a riveting account of talking to his children about the shooting in January, also declined to comment to Yahoo News on gun control.
"I appreciate your position as a journalist," Eller wrote in an email, "but I do not wish to wade into the gun debate at this time."
There were many signs that the town has returned to some normalcy. At Starbucks, for instance, a group of caffeinated teen girls texted away on iPhones instead of studying flash cards. A copy of Monday's Daily News—with Newtown gunman Adam Lanza on the cover—was left unread on a nearby table.


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The gems were taken from the vault at a branch of the Dutch bank ABN Ambro in the city of Antwerp, where half the world's diamonds are traded.
A composite picture of the suspect has been issued by police.
They said he was a regular customer at the bank and appeared to have prepared for the theft for over a year.
The robbery was discovered a week ago, but has only now been made public.
Several safes were broken into and the haul included both cut and rough diamonds, police said.
Most Popular Now | 56,514 people are reading stories on the site right now.";