Probing of Exosites Leads to Novel Inhibitor Scaffolds of HCV NS3/4A Proteinase

Hepatitis C is a treatment-resistant disease affecting millions of people worldwide. The hepatitis C virus (HCV) genome is a single-stranded RNA molecule. After infection of the host cell, viral RNA is translated into a polyprotein that is cleaved by host and viral proteinases into functional, structural and non-structural, viral proteins. Cleavage of the polyprotein involves the viral NS3/4A proteinase, a proven drug target. HCV mutates as it replicates and, as a result, multiple emerging quasispecies become rapidly resistant to anti-virals, including NS3/4A inhibitors.To circumvent drug resistance and complement the existing anti-virals, NS3/4A inhibitors, which are additional and distinct from the FDA-approved telaprevir and boceprevir α-ketoamide inhibitors, are required. To test potential new avenues for inhibitor development, we have probed several distinct exosites of NS3/4A which are either outside of or partially overlapping with the active site groove of the proteinase. For this purpose, we employed virtual ligand screening using the 275,000 compound library of the Developmental Therapeutics Program (NCI/NIH) and the X-ray crystal structure of NS3/4A as a ligand source and a target, respectively. As a result, we identified several novel, previously uncharacterized, nanomolar range inhibitory scaffolds, which suppressed of the NS3/4A activity in vitro and replication of a sub-genomic HCV RNA replicon with a luciferase reporter in human hepatocarcinoma cells. The binding sites of these novel inhibitors do not significantly overlap with those of α-ketoamides. As a result, the most common resistant mutations, including V36M, R155K, A156T, D168A and V170A, did not considerably diminish the inhibitory potency of certain novel inhibitor scaffolds we identified.Overall, the further optimization of both the in silico strategy and software platform we developed and lead compounds we identified may lead to advances in novel anti-virals

The device for testing the spring mechanisms to a drive of medium voltage switch-disconnectors

Integracja zaawansowanych przetworników pomiarowych, technik cyfrowego sterowania napędami oraz metod bezpiecznego programowania pozwala automatyzować nie tylko procesy wytwórcze, lecz także złożone i czasochłonne procedury testowania i badania wyrobów. Przykładem takiej zintegrowanej aplikacji pomiarowej jest opracowane w PIAP urządzenie testujące mechanizmy sprężynowe do napędu rozłączników średnich napięć, które zostało wdrożone w 2009 r. w firmie ABB Oddział w Przasnyszu. Urządzenie zdejmuje charakterystyki momentu obrotowego w funkcji kąta obrotu, przy załączaniu i wyłączaniu mechanizmu sprężynowego. Uzyskano dokładność i wydajność testowania nieosiągalne przy manualnym zdejmowaniu charakterystyk. Wyeliminowano także zagrożenia dla obsługi przy operowaniu mechanizmami sprężynowymi kumulującymi znaczne energie.The integration of advanced measuring converters, techniques of digital control for drives and methods of safe programming permits to automate not only productive processes, but also complex and time-consuming testing procedures and investigations of products. The example of such integrated measuring application is worked out in PIAP the device for testing spring mechanisms to a drive of medium voltage switch-disconnectors which were implemented in 2009 in the division of ABB in Przasnysz. The device takes off torque characteristics as function of turn angle, when enclosing and disengaging of spring mechanism. The accuracy and testing rate obtained were unattainable using characteristics taken off manually. Risks to service staff by operating the spring mechanisms also were eliminated

Design of CD40 Agonists and Their Use in Growing B Cells for Cancer Immunotherapy

CD40 stimulation has produced impressive results in early-stage clinical trials of patients with cancer. Further progress will be facilitated by a better understanding of how the CD40 receptor becomes activated and the subsequent functions of CD40-stimulated immune cells. This review focuses on two aspects of this subject. The first is the recent recognition that signaling by CD40 is initiated when the receptors are induced to cluster within the membrane of responding cells. This requirement for CD40 clustering explains the stimulatory effects of certain anti-CD40 antibodies and the activity of many-trimer, but not one-trimer, forms of CD40 ligand (CD40L, CD154). The second topic is the use of these CD40 activators to expand B cells (“CD40-B cells”). As antigen-presenting cells (APCs), CD40-B cells are as effective as dendritic cells, with the important difference that CD40-B cells can be induced to proliferate in vitro whereas DCs proliferate poorly if at all. As a result, the use of CD40-B cells as antigen-presenting cells (APCs) promises to streamline the generation of anti-tumor CD8+ T cells for the adoptive cell therapy (ACT) of cancer