The flavivirus polymerase as a target for drug discovery.

Flaviviruses are emerging pathogens of increasingly important public health concern in the world. For most flaviviruses such as dengue virus (DENV) and West Nile virus (WNV) neither vaccine nor antiviral treatment is available. The viral RNA-dependent RNA polymerase (RdRp) non-structural protein 5 (NS5) has no equivalent in the host cell and is essential for viral replication. Here, we give an overview of the current knowledge regarding Flavivirus RdRp function and structure as it represents an attractive target for drug design. Flavivirus RdRp exhibits primer-independent activity, thus initiating RNA synthesis de novo. Following initiation, a conformational change must occur to allow the elongation process. Structure-function studies of Flavivirus RdRp are now facilitated by the crystal structures of DENV (serotype 3) and WNV RdRp domains. Both adopt a classic viral RdRp fold and present a closed pre-initiation conformation. The so-called priming loop is thought to provide the initiation platform stabilizing the de novo initiation complex. A zinc-ion binding site at the hinge between two subdomains might be involved in opening up the RdRp structure towards a conformation for elongation. Using two different programs we predicted common potential allosteric inhibitor binding sites on both structures. We also review ongoing approaches of in vitro and cell-based screening programs aiming at the discovery of nucleosidic and non-nucleosidic inhibitors targeting Flavivirus RdRps.

Rectal augmentation: short- and mid-term evaluation of a novel procedure for severe fecal urgency with associated incontinence.

BACKGROUND: Rectal augmentation (RA) with or without electrically stimulated gracilis neosphincter (ESGN) was developed to address the physiologic and anatomic abnormalities present in a subset of patients with incapacitating fecal urgency and associated urge fecal incontinence (UFI). This study evaluated the short- and medium-term clinical and physiologic results. METHODS: Eleven patients with fecal urgency and UFI underwent RA, 6 with concomitant ESGN formation. Patients were evaluated preoperatively, and at a median of 12.5 and 54 months after surgery. RESULTS: At 4.5 years, 7/11 patients had avoided stoma construction. Symptoms recurred leading to permanent stoma formation in 1 patient, whereas one other developed evacuatory difficulty with overflow incontinence. Median ability to defer defecation improved from seconds preoperatively to 10 minutes at 1 year (P = 0.0002), and 15 minutes at 4.5 years (P = 0.002). Median Wexner incontinence scores improved from 15 preoperatively to 3 at 1 year (P = 0.002), and 4 at 4.5 years (P = 0.02). At 1 year, 2 of the rectal sensory thresholds (DDV: P = 0.008; MTV: P = 0.008) and compliance were normalized (P = 0.008), whereas at 4.5 years, all sensation thresholds improved (FCS: P = 0.002; DDV: P = 0.002; MTV: P = 0.002), but changes in compliance were not significant. CONCLUSION: RA with or without ESGN improved reported symptoms and normalized rectal sensation. Improvements were sustained in the medium term. The procedure had no associated morbidity or mortality, and should be considered in the surgical management of a select group of patients presenting with severe urgency and UFI.