Growth kinetics of rLaSota and rBanj FPCS mutant viruses in 9-day-old embryonated SPF chicken eggs.

<p>Two hundred μL of 100 TCID₅₀ of each virus in PBS was injected into the allantoic cavity of ten 9-day-old embryonated SPF chicken eggs. Three days after incubation at 37°C, the allantoic fluids was harvested and titrated by hemagglutination (HA) assay. The bars are the means of HA titers. Error bars indicate standard error of the mean.</p

Pathogenicity of rBanj FPCS mutant viruses in 1-day-old SPF chicks.

<p>Pathogenicity of rBanj FPCS mutant viruses in 1-day-old SPF chicks.</p

Virus titers and tissue tropism of rBanj FPCS mutant viruses in 1-day-old chicks following oculo-nasal inoculation.

<p>Tissue samples from brain, lungs, trachea, and spleen of 3 chickens (n  =  3) from each indicated virus group were collected on day 3 post infection, and virus titers were determined by TCID₅₀ assay. The mean virus titers for each tissue sample from 3 chickens are shown. Error bars indicate standard error of the mean.</p

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Cytopathogenicity of rBanj FPCS mutants in DF-1 cells.

<p>The cells were infected at an MOI of 1 with each of the recombinant viruses. After 3 days, the cytopathic effects (CPE) of each virus infected monolayer was examined under microscope (10X). Ten percent fresh chicken embryo allantoic fluid was used as the source of exogenous protease.</p

Growth kinetics of rLaSota and rBanj FPCS mutant viruses in DF-1 cells.

<p>Cells were infected at an MOI of 0.001 of each virus and the cell culture supernatant was collected at 8 h intervals for 64 h. All virus titers are expressed as mean log₁₀ TCID₅₀/mL ± SEM (standard error of the mean).</p

Effect of fusion protein cleavage site sequence on generation of a genotype VII Newcastle disease virus vaccine - Fig 8

<p><b>Induction of NDV-specific serum antibodies in chickens in response to vaccination with rBanj FPCS mutant viruses, determined by HI (A and B) and serum neutralization assays (C and D).</b> (A and B) Twenty 1-day-old chicks per group were inoculated with 100 μL of (1x10⁵ PFU) virus via oculo-nasal route. Serum samples were collected at 1, 2, and 3 weeks post inoculation. NDV-specific antibodies were determined by hemagglutination inhibition (HI) assay sing 4 HA units of rBanj-LaSota (A) and rLaSota (B). (C and D) The serum neutralizing titers of the vaccinated birds against rBanj-LaSota-eGFP (C) and rLaSota-eGFP (D) viruses are shown. Two-fold serial dilutions of complement-inactivated serum samples were made in a 96-well plate and incubated with 1x10³ TCID₅₀ of the recombinant virus at 37°C for 1 h. After incubation at 37°C for 48 h, the cells were washed with PBS and fixed using 4% paraformaldehyde. The fluorescence intensity was measured using a micro plate reader (Tecan, Infinite® M1000) at 490 nm. The neutralization titer was defined as the reciprocal of the highest serum dilution that resulted in 50% reduction in mean eGFP fluorescence.</p

Serum antibody responses against APMV serotypes 1–9 in infected mice^a.

<p>All the values are averages from three independent experiments.</p>a<p>Mice in groups of 3 were inoculated as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0016776#pone-0016776-t001" target="_blank">Table 1</a>. Serum samples were collected before inoculation and 14 dpi.</p>b<p>The hemagglutination inhibition (HI) titer is expressed as the reciprocal of the highest serum dilution causing complete inhibition of four HA units of NDV.</p><p>-  = not detected.</p

Virus isolation from the indicated tissue harvested from mice 3 dpi with APMV serotypes 1 to 9^*.

*<p>Mice in groups of 3 were inoculated with 50 ìl of allantoic fluid containing 2⁷ HA units of each APMV serotype except serotype 5, which contained 3×10³ PFU/ml of cell culture harvested virus. The control group was inoculated with normal allantoic fluid. Tissues were harvested 3 dpi and homogenized, and clarified supernatant fluid was inoculated into 9-day-old embryonated eggs and tested for virus 4 days later by HA assay.</p><p>+  = each + indicates isolation of virus from one mice.</p><p>-  = no virus was isolated.</p><p>ND = not detected.</p

HIV-1 Env-specific CD4+ (panel A) and CD8+ (panel B) T cell response.

<p>Mice in groups of 6 were immunized with 10⁵ PFU/ml of the indicated rNDV by the i.n. route on days 0 and 14. On day 56, splenocytes were isolated, stimulated with a pool of overlapping Env peptides, and processed for intracellular cytokine staining for IFN-γ and CD4 and CD8.</p