Journal: Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases

Article Title: Nausea, Vomiting, and Noninflammatory Diarrhea

doi: 10.1016/B978-1-4557-4801-3.00100-4

Figure Lengend Snippet: Etiology of Traveler's Diarrhea *

Article Snippet: The first rotavirus vaccine approved for use in the United States was the rhesus-human reassortant rotavirus tetravalent vaccine (RRV-TV; Rotashield, Wyeth-Lederle, Madison, NJ).

Techniques:

Journal: PLoS Pathogens

Article Title: Cell–cell fusion induced by reovirus FAST proteins enhances replication and pathogenicity of non-enveloped dsRNA viruses

doi: 10.1371/journal.ppat.1007675

Figure Lengend Snippet: ( A ) Enhancement of viral replication by recombinant PRV FAST-p10 protein. Vero cells were transfected with FAST-p10 expression vector at the indicated times either before or after infection with a FAST-p10-deficient PRV mutant (rsMB-ΔFAST). Infectious virus titers in cell lysates at 16 h post infection were determined. Data are expressed as means ± SD ( n = 3) and were statistically analyzed using the t -test. ( B ) Whole-cell lysates of Vero cells transfected with FAST-p10 expression vector or empty vector 2 h before infection with rsMB-ΔFAST were prepared, and viral sigmaC protein was detected by western blotting. ( C ) Vero cells were transfected with FAST-p10 expression vector or empty vector (0.25–2.0 μg/well) 2 h before infection with wild-type PRV (rsMB) or rsMB-ΔFAST. Infectious virus titers in cell lysates at 16 h post infection were determined. Data are expressed as means ± SD ( n = 3) and were statistically analyzed using the t -test. ( D ) Efficiency of syncytium formation by different members of the FAST protein family. Vero cells were transfected with recombinant expression vectors (0.5 μg/well) for PRV FAST-p10, avian orthoreovirus (ARV) FAST-p10, reptilian orthoreovirus (RRV) FAST-p14, or baboon orthoreovirus (BRV) FAST-p15, or with empty vector. At 16 h post infection, cells were fixed and numbers of nuclei per syncytium were counted ( n = 5–33). * indicates p < 0.05 (Dunn’s multiple-comparison test). ( E ) Vero cells were transfected with expression vectors for PRV FAST-p10, ARV FAST-p10, RRV FAST-p14, or BRV FAST-p15, or with empty vector (0.1–2.0 μg/well) 2 h before infection with rsMB-ΔFAST. Infectious virus titers in cell lysates at 16 h post infection were determined. Data are expressed as means ± SD ( n = 4). Viral titers in cells with 2.0 μg/well plasmid transfections were statistically compared with titers in mock-transfected samples. * indicates p < 0.05. ( F ) QT6 cells were transfected with 2 μg of PRV FAST-p10 expression vector or empty vector, followed by infection with ARV at a MOI of 0.001 PFU/cell. Infectious virus titers in cell lysates at 16 h post infection were determined and statistically analyzed using the t -test. Data are expressed as means ± SD ( n = 3). ( G ) Time course of syncytium formation by PRV FAST-p10 and modified recombinant Sendai virus (SeV) F (Fc) proteins. Vero cells were transfected with expression vectors either for FAST-p10 or for SeV Fc and SeV HN (0.5 μg each/well). At indicated times post transfection, cells were fixed and numbers of nuclei involved per syncytium were counted. Data are presented as a box plot ( n = 7–30). ( H ) Enhancement of viral replication by recombinant SeV Fc protein. Vero cells were transfected with expression vectors for SeV Fc and SeV HN proteins (1 μg each/well) 2 h before infection with rsMB-ΔFAST at a MOI of 0.001 PFU/cell. At 16 h post infection, infectious virus titers in the cell lysates were determined. Data are expressed as means ± SD ( n = 3) and were statistically analyzed using the t -test.

Article Snippet: BRV p15 gene (AF406787) and RRV p14 gene (AY238887) were synthesized (Eurofins Genomics) and cloned into pCAGGS plasmid to create pCAG-BRVp15-FAST and pCAG-RRVp14-FAST, respectively.

Techniques: Recombinant, Transfection, Expressing, Plasmid Preparation, Infection, Mutagenesis, Virus, Western Blot, Comparison, Modification