Treatment of malignant gliomas with a replicating adenoviral vector expressing herpes simplex virus-thymidine kinase

We evaluated the interaction between oncolytic, replication-competent adenoviral vectors and the herpes simplex virus-1 thymidine kinase (HSV1-tk) gene/ganciclovir (GCV) suicide system for the treatment of malignant gliomas. We constructed a panel of replication-competent adenoviral vectors in which the luciferase (IG.Ad5E1(+). E3Luc) or HSV1-tk gene (IG.Ad5E1(+).E3TK) replace the M(r) 19,000 glycoprotein (gp19K) coding sequence in the E3 region. IG.Ad5E1. IG.Ad5.ClipLuc and IG.AdApt.TK are E1-deleted viruses that contain the luciferase or the HSV1-tk gene in the former E1 region driven by the human cytomegalovirus promoter. IG.Ad5.Sarcoma 1800HSA.E3Luc contains an irrelevant gene in the E1 region, whereas the gp19K coding sequence in the E3 region is replaced by the luciferase gene as in the replicating virus IG.Ad5E1(+).E3Luc. For in vitro experiments, we used a panel of human glioma cell lines (U87 MG, T98G, A172, LW5, and U251), a rat gliosarcoma cell line (9 L), and human lung (A549) and prostate carcinoma (P3) cell lines. In vitro, GCV sensitivity (10 microg/ml) was studied in U87 MG cells after infection at a multiplicity of infection of 1 and 10. A s.c. U87 MG glioma xenograft model was established in NIH-bg-nu-xid mice. Tumors of 100-150 mm(3) were treated with a single injection of adenovirus 10(9) IU suspended in 100 microl of PBS, and GCV 100 mg/kg was administered i.p. twice daily for 7 days. The cytopathic effect of all three replication-competent adenoviral vectors was similar to the cytopathic effect of wild-type adenovirus 5 on all human cell lines tested, indicating that deletion of the E3 gp19K sequences did not affect the oncolytic effect of the vectors. In vitro, luciferase expression was the same for both E1-deleted vectors (IG.Ad5.ClipLuc and IG.Ad5.Sarcoma 1800HSA.E3Luc), demonstrating the strength of the internal E3 promoter even in the absence of E1A. However, in vitro expression levels obtained with replication-competent IG.Ad5E1(+). E3Luc were 3 log higher (allowing infection with a

JUSTICE AS BEAUTY-IN-ACTION? INSIGHTS FROM HANS URS VON BALTHASAR’S AESTHETICS AND DRAMATICS

This essay explores the relation between beauty and justice by turning to the thought of the Swiss Catholic theologian Hans Urs von Balthasar. It begins by giving an exposition of Balthasar’s theological aesthetics, as developed in his work The glory of the Lord, which shows how, for Von Balthasar, earthly beauty participates in, and expresses something of God’s divine glory and reaches its apex in the revelation of the beautiful form of Jesus Christ. This is then followed by an exposition of Von Balthasar’s theological dramatics, as developed in his work Theo-drama, which shows how, for Von Balthasar, this beautiful form of Christ is not merely a static image, icon, or artwork but, in fact, a dynamic event, a dramatic act, an embodied performance which reveals to us, along with God’s glory and beauty, God’s unbounded goodness. The essay subsequently turns to questions of justice (in light of Von Balthasar’s understanding of the relation between beauty and goodness), and ultimately argues that, according to Von Balthasar’s thought, justice can be viewed as a form of beauty-in-action that asks to be performed in the world

Gene transfer of the urokinase-type plasminogen activator receptor-targeted matrix metalloproteinase inhibitor TIMP-1.ATF suppresses neointima formation more efficiently than tissue inhibitor of metalloproteinase-1

Proteases of the plasminogen activator (PA) and matrix metalloproteinase (MMP) system play an important role in smooth muscle cell (SMC) migration and neointima formation after vascular injury. Inhibition of either PAs or MMPs has previously been shown to result in decreased neointima formation in vivo. To inhibit both protease systems simultaneously, a novel hybrid protein, TIMP-1.ATF, was constructed consisting of the tissue inhibitor of metalloproteinase-1 (TIMP-1) domain, as MMP inhibitor, linked to the receptor-binding amino terminal fragment (ATF) of urokinase. By binding to the u-PA receptor this protein will not only anchor the TIMP-1 moiety directly to the cell surface, it will also prevent the local activation of plasminogen by blocking the binding of urokinase-type plasminogen activator (u-PA) to its receptor. Adenoviral expression of TIMP-1.ATF was used to inhibit SMC migration and neointima formation in human saphenous vein segments in vitro. SMC migration was inhibited by 65% in Ad.TIMP-1.ATF-infected cells. Infection with adenoviral vectors encoding the individual domains, Ad.TIMP-1 and Ad.ATF, reduced migration by 32% and 52%, respectively. Neointima formation in saphenous vein organ cultures infected with Ad.TIMP-1.ATF was inhibited by 72% compared with 42% reduction after Ad.TIMP-1 infection and 34% after Ad.ATF infection. These data show that binding of TIMP-1.ATF hybrid protein to the u-PA receptor at the cell surface strongly enhances the inhibitory effect of TIMP-1 on neointima formation in human saphenous vein cultures

New molecular surrogate assay for genotoxicity assessment (SAGA)

Stemcel biology/Regenerative medicine (incl. bloodtransfusion

Effects of prepartum and postpartum bolus injections of trace minerals on performance of beef cows and calves grazing native range

Adequate dietary intakes of trace minerals are thought necessary to maximize cow reproduction, calf health, and calf performance. Diets grazed by beef cattle are generally deficient to marginal in copper (Cu), manganese (Mn), selenium (Se), and zinc (Zn) concentrations; therefore, these trace minerals are usually added to the diet in supplement form. The most widely used means of trace-mineral supplementation for grazing cattle is selffed, salt-based, loose mineral supplements. Although cattle do not balance their mineral needs when consuming a self-fed mineral supplement, usually no other practical way of supplying mineral needs exists under grazing conditions. The greatest limitation to using self-fed mineral supplements is variation in animal intake. More direct methods of mineral supplementation include adding minerals to drinking water or feed, oral drenching, ruminal boluses, and injection. Variation in mineral intake is reduced relative to self-fed supplementation, and the additional labor requirement and expense are relatively small. Delivery of supplemental trace minerals using an injectable solution may be a more reliable means of achieving adequate trace-mineral status than using self-fed, salt-based, loose mineral supplements. Bolus injections of trace minerals have been associated with improved average daily gain, feed efficiency, feed intake, or health status of beef calves fed in confinement; however, trace mineral delivery methods of this type have not been fully evaluated with respect to performance of beef cows and suckling calves. The objective of our study was to evaluate the effects of pre- and postpartum bolus injections of a trace mineral solution on beef cow reproductive performance, body weight change, and body condition score change, as well as performance of suckling calves

Imaging expression of adenoviral HSV1-tk suicide gene transfer using the nucleoside analogue FIRU.

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Expression of aberrantly glycosylated tumor mucin-1 on human DC after transduction with a fiber-modified adenoviral vector

Background: DC-presenting tumor Ag are currently being developed to be used as a vaccine in human cancer immunotherapy. To increase chances for successful therapy it is important to deliver full-length tumor Ag instead of loading single peptides. Methods: In this study we used a fiber-modified adenoviral vector (rAd5F35) containing full-length tumor Ag cDNA to transduce human monocyte (Mo)-derived DC in vitro. Cells were efficiently transduced and survived for at least 3 days after adenoviral transduction. Phenotype and function after maturation of Mo-DC were not impaired by infection with adenovirus particles. Expression of the tumor-associated Ag mucin-1 (MUC1) was detected using MAb defining different MUC1 glycoforms. Results: Non-transduced mature Mo-DC express endogenous MUC1 with normal glycosylation. After transduction with the rAd5F35-MUC1 adenoviral vector, Mo-DC also expressed MUC1 with tumor-associated glycosylation (Tn and T glycoforms), although no changes in mRNA levels of relevant glycosyltransferases could be demonstrated. Discussion: The presence of aberrantly glycosylated MUC1 may influence Ag presentation of the tumor glycoforms of MUC1 to immune cells, affecting tumor cell killing. These findings could be highly relevant to developing strategies for cancer immunotherapy based on DC vaccines using MUC1 as tumor Ag

Immune control of an SIV challenge by a T-cell-based vaccine in rhesus monkeys

A recombinant adenovirus serotype 5 (rAd5) vector-based vaccine for HIV-1 has recently failed in a phase 2b efficacy study in humans. Consistent with these results, preclinical studies have demonstrated that rAd5 vectors expressing simian immunodeficiency virus (SIV) Gag failed to reduce peak or setpoint viral loads after SIV challenge of rhesus monkeys (Macaca mulatta) that lacked the protective MHC class I allele Mamu-A*01 (ref. 3). Here we show that an improved T-cell-based vaccine regimen using two serologically distinct adenovirus vectors afforded substantially improved protective efficacy in this challenge model. In particular, a heterologous rAd26 prime/rAd5 boost vaccine regimen expressing SIV Gag elicited cellular immune responses with augmented magnitude, breadth and polyfunctionality as compared with the homologous rAd5 regimen. After SIVMAC251 challenge, monkeys vaccinated with the rAd26/rAd5 regimen showed a 1.4 log reduction of peak and a 2.4 log reduction of setpoint viral loads as well as decreased AIDS-related mortality as compared with control animals. These data demonstrate that durable partial immune control of a pathogenic SIV challenge for more than 500 days can be achieved by a T-cell-based vaccine in Mamu-A*01-negative rhesus monkeys in the absence of a homologous Env antigen. These findings have important implications for the development of next-generation T-cell-based vaccine candidates for HIV-1. ©2009 Macmillan Publishers Limited. All rights reserved