Severe Acute Respiratory Syndrome Coronavirus Envelope Protein Regulates Cell Stress Response and Apoptosis

Severe acute respiratory syndrome virus (SARS-CoV) that lacks the envelope (E) gene (rSARS-CoV-ΔE) is attenuated in vivo. To identify factors that contribute to rSARS-CoV-ΔE attenuation, gene expression in cells infected by SARS-CoV with or without E gene was compared. Twenty-five stress response genes were preferentially upregulated during infection in the absence of the E gene. In addition, genes involved in signal transduction, transcription, cell metabolism, immunoregulation, inflammation, apoptosis and cell cycle and differentiation were differentially regulated in cells infected with rSARS-CoV with or without the E gene. Administration of E protein in trans reduced the stress response in cells infected with rSARS-CoV-ΔE or with respiratory syncytial virus, or treated with drugs, such as tunicamycin and thapsigargin that elicit cell stress by different mechanisms. In addition, SARS-CoV E protein down-regulated the signaling pathway inositol-requiring enzyme 1 (IRE-1) of the unfolded protein response, but not the PKR-like ER kinase (PERK) or activating transcription factor 6 (ATF-6) pathways, and reduced cell apoptosis. Overall, the activation of the IRE-1 pathway was not able to restore cell homeostasis, and apoptosis was induced probably as a measure to protect the host by limiting virus production and dissemination. The expression of proinflammatory cytokines was reduced in rSARS-CoV-ΔE-infected cells compared to rSARS-CoV-infected cells, suggesting that the increase in stress responses and the reduction of inflammation in the absence of the E gene contributed to the attenuation of rSARS-CoV-ΔE

Evaluation of cytotoxicity and degree of conversion of orthodontic adhesives over different time periods

As new orthodontic resin adhesives continue to be marketed, rapid and sensitive tests for examining their toxic effects at the ' cell and tissue level ' are needed because patient safety has been identifi ed as a legal concept. The objective of the present study was to evaluate the cytotoxicity and degree of monomer conversion of orthodontic adhesives over different time periods. Seven adhesives: Transbond® XT, Transbond® Color Change, Quick Cure, EagleBond, Orthobond®, Fill Mágic® and Biofix® were evaluated for their cytotoxicity in L929 fibroblastic cells and for their degree of monomer conversion over different time periods. Three control groups were also analysed: Positive control (C+), consisting of Tween 80 cell detergent; Negative control (C-), consisting of PBS; and cell control (CC), consisting of cells exposed to any material. The dye-uptake technique that involves the absorption of a neutral red dye in viable cells was used for the cytotoxicity evaluation and the degree of conversion was evaluated using spectroscopy with infrared. The results showed the cytotoxicity of the adhesives at 24, 48, 72 and 168 hours. At these times, the viability values presented for these materials were statistically different from the groups CC and C- (p < 0.05). At 168 hours, all the groups showed low cytotoxicity with high cell viability and with no statistical difference with the groups CC and C- (P > 0.05). In the monomer conversions there was a percentage increase of monomer conversion from 24 to 72 hours. A direct correlation could be observed between cytotoxicity and monomer conversions. From this work it can be concluded that all adhesives evaluated are cytotoxic at the times of 24, 48 and 72 hours. Monomers continued conversion even after photopolymerization had stopped