Studies on the loss of gloss of shellac and polyurethane finishes exposed to UV (AOP Paper)

<p>Wood finishes protect the surface of wood from external agents, enhance its looks and improve its gloss (luster). On constant external exposure, UV rays gradually degrade the film coating resulting in loss of gloss. In this study, two commonly used finishes namely spirit shellac and polyurethane finish were used to investigate the pattern of loss of gloss due to UV interference. Two coatings of polyurethane (PU) and spirit shellac finish were applied on the surface of eucalyptus samples. The gloss levels of these and unfinished samples were monitored for different times of exposure of them to UV light. Gloss was measured at 600 gloss head using a Tri micro gloss meter regularly for 20 hours durations of UV exposure. Observations and analysis revealed that the natural gloss of uncoated samples of eucalyptus were least affected on UV exposure with only 8.3%-10% loss in gloss. The PU and shellac coated surfaces also showed very little reduction in gloss (6.9%-15.4%) most of which happened in the first 40 hours of exposure. Thus both the finishes in this study were found effective to a good extent in maintaining the gloss of the finished surface against exposure to UV light.</p

RealTime PCR analysis of 293T cells transfected with LMP1 or ΔLMP1-MAVS.

<p>293T cells were transfected with pcDNA3.1, LMP1 plasmid, or ΔLMP1-MAVS plasmid and total RNA isolated following 36-hour culture. Normalized expression was determined relative to the pcDNA3.1 control.</p

ΔLMP1-MAVS enhances anti-Gag immune responses as an Ad5 viral vector vaccine adjuvant.

<p>BALB/c mice were left untreated or vaccinated with a combination of Ad5-Gag and either Ad5-GFP (control), Ad5-ΔLMP1-MAVS, or Ad5-LMP1. Two weeks following vaccination, mice were challenged with vaccinia-gag virus. Vaccinia titers were measured from ovaries after 5 days. NT: no treatment.</p

Gene array analysis of transfected 293T cells and primary CD4+ T cells cultured with 293T supernatant.

<p>Three independent wells of 293T cells were transfected with pcDNA3.1 empty vector or ΔLMP1-MAVS plasmid and total RNA isolated 36 hours later. Primary CD4+ T cells from 3 independent donors were cultured with 293T supernatant (collected 24 hours following pcDNA3.1 or ΔLMP1-MAVS transfection). Total CD4+ T cell RNA was isolated 36 hours later. (A) Venn Diagrams of the number of probe sets upregulated and down-regulated (>2-fold change) by ΔLMP1-MAVS. (B) Differential gene expression of 293T cells and CD4+ T cells. (C) List of genes upregulated by ΔLMP1-MAVS in both 293T cells and CD4+ T cells. Fold-change and P-values for each probe set are shown for CD4+ T cells following ΔLMP1-MAVS treatment. (D) List of genes upregulated by CD4+ T cells but not 293T cells. Fold-change between pcDNA3.1 and pΔLMP1-MAVS and P-values for each probe set are shown. (E) Gen Go networks analysis of pathways significantly upregulated by ΔLMP1-MAVS in transfected 293T cells, or CD4+ T cells cultured with ΔLMP1-MAVS transfected 293T supernatant.</p

Beta-interferon mediates inhibition of HIV-1 BaL replication.

<p>(A) The relative level of HIV-1 BaL strain viral replication in TZM-bl cells was measured following incubation with supernatant from 293T cells transfected with either pcDNA3.1 or pΔLMP1-MAVS plasmid, combined with 60 μg of isotype control antibody, anti-IFN-β antibody, or anti-IFN-α antibody. (B) Human CD4+ T cells were infected with HIV-1 BaL in the presence of increasing concentrations of interferon-α and compared to infection in the presence of 293T supernatant following transfection with either pcDNA3.1 or pΔLMP1-MAVS plasmid. (C) Supernatant from 293T cells transfected with pcDNA3.1 GFP, LMP1, or ΔLMP1-MAVS plasmid was assayed for IFN-α and IFN-β secretion by ELISA. NT: no treatment.</p

Inhibition of HIV and SIV viral infection of TZM-bl and primary human CD4+ T cells.

<p>The relative level of viral replication in TZM-bl cells was measured following incubation with supernatant from 293T cells transfected with pcDNA3.1 vector expressing GFP (control), LMP1, or ΔLMP1-MAVS plasmid. (A) Infection with HIV-1 BaL strain. (B) Infection with VSV-G pseudotyped single cycle SIV. (C) CD4+ T cells were isolated from a healthy donor by negative selection, activated, and cultured with 293T supernatant for 24 hours. Cells were then washed and infected with HIV-1 BaL at an MOI or 0.1 or 1. The concentration of p24 was measured 6 days later by ELISA assay. *p<0.05, **p<0.01, ***p<0.001.</p

Exosome-depleted 293T supernatant inhibits HIV and SIV replication.

<p>293T cells were transfected with pcDNA3.1 plasmids encoding GFP, LMP1, or ΔLMP1-MAVS and supernatant was isolated. Supernatant was depleted of exosomes by ultracentrifugation. (A) TZM-bl cells were cultured with isolated exosomes and infected with increasing concentrations of HIV-1 BaL. (B) TZM-bl cells were cultured with exosome-depleted supernatant, followed by infection with HIV-1 BaL. (C) TZM-bl cells were cultured with exosome-depleted supernatant, followed by infection with VSV-G pseudotyped single cycle SIV. *p<0.05, **p<0.01, ***p<0.001.</p

Biological activity of Ad5-LMP1.

<p><b>(A, B)</b> 293T cells were transfected with either NF-κB <b>(A)</b> or IFN-β <b>(B)</b> firefly luciferase reporter plasmid together with pRL-TK and either LMP1 or control expression plasmids. As positive controls, pcDNA3.1-FLAG-TRAF6 and pcDNA3.1-ΔRIG-I were used. 36-48h later, luciferase activity was measured in total cell lysate. <b>(C)</b> Viral stocks of Ad5-LMP1 were serially diluted and added to 293-SEAP reporter cells in triplicate, in the presence or absence of doxycycline. After 36h, secreted alkaline phosphatase was measured. Data represent typical results from three independent experiments.</p

Ad5-LMP1 enhanced maturation, cytokine production of human monocyte-derived dendritic cells.

<p>Human monocytes were isolated from buffy coat and cultured in the presence of GM-CSF and IL-4 for 5 days to generate DC. On day 5, cells were infected with Ad5-LMP1 or Ad5-GFP at an MOI of 100 in the presence of doxycycline. Mimic cytokine maturation cocktail was added on day 5 as a positive control. <b>(A)</b> Cells were analyzed by flow cytometry and representative forward vs. side scatter plots are displayed. <b>(B)</b> Images of DC were taken on day 7 before harvesting for maturation staining. Scale bar equals 100μm. <b>(C)</b> Representative gating strategy used to determine maturation of MDDC. <b>(D)</b> DC were stained for maturation markers, and analyzed by flow cytometry. <b>(E)</b> Supernatant was collected every 12 hours for 2 days and cytokine measured by CBA. TNF-α, IL-6 and IL-1β levels for Mimic sample may represent residual protein from the Mimic cocktail. Data represent results from three independent experiments.</p

Ad5-LMP1 DC reduced viral titers of vaccinia-Gag in a prophylactic DC vaccination model.

<p>BMDC were transduced with Ad5-Gag and matured with either Ad5-LMP1 or Mimic in the presence of doxycycline. Mice were vaccinated with 1x10⁵ DC injected intradermally into the flank. Mice were given doxycycline in drinking water from the time of injection. 4 weeks post-vaccination, mice were challenged with 1x10⁷ viral particles vaccinia-Gag. Five days following challenge, ovaries were assayed for viral titer. Data combine the results from two independent experiments (n = 9 mice per group).</p