(c) 2008 Elsevier Ireland Ltd. All rights reserved.”
“Real-time RT-PCR and
Northern blot are employed for the measurement of HCV RNA but suffer from multiple purification steps, high cost, and relatively large variability. In this study, a hybridization method for HCV RNA detection is described. This method does not need RNA purification, and is sensitive enough to detect HCV RNA present in replicon cellular lysates harvested from a single well of a 96-well plate. Fixation of RNA by UV cross-linking is crucial for this sensitivity. A linear relationship exists between hybridization signal and cell density ranging from 10(5) to as few as 300 cells per well. The signal-to-background
ratio is greater than 40 and the Z factor is above 0.7. Using several known anti-HCV agents, dose-response curves and EC50 values generated find more from hybridization were similar to those obtained from a luciferase assay. This method has been successfully applied to replicons of different HCV subtypes and hepatitis B virus in our laboratory. In summary, this hybridization assay is sensitive, highly reproducible, easy to handle, and a valuable tool for antiviral drug discovery. (C) 2008 Elsevier B.V. All rights reserved.”
“Herpes simplex virus type-1 (HSV-1) is a neurotropic virus with significant potential as a viral vector D-malate dehydrogenase for central nervous system (CNS) PSI-7977 molecular weight gene therapy. This study provides visual evidence that recombinant green fluorescent protein (GFP)-expressing HSV-1 travel down dendrites in differentiated P19 neuronal-like cells to efficiently reach the soma. The virus also promotes cytoskeletal rearrangements which facilitate viral spread in vitro, including often dramatic increases in dendritic filopodia. Viral movements, cell infection and filopodia induction were each reduced with the actin polymerization inhibitor cytochalasin D, suggesting the involvement of the actin cortex in these processes. The observation of neural cytoskeletal
reorganization in response to HSV-I may shed light on the mechanisms by which acute viral infection associated with herpes encephalitis produces cognitive deficits in patients. (c) 2008 Elsevier Ireland Ltd. All rights reserved.”
“Prior to PCR amplification, uracil-DNA glycosylase (UNG) can be incorporated to prevent amplicon contamination. Melting temperature (T) analysis of herpes simplex virus (HSV) hybridization probes was used to demonstrate a concentration-dependent decrease in T-m values when heat stabile or heat labile UNG was added. (C) 2008 Elsevier B.V. All rights reserved.”
“Patients with schizophrenia have impairment in cognitive-emotional interaction that is reflected in delusion formation and poor social functioning.
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