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Abstract Background The RNA interference (RNAi) pathway acts as an innate antiviral immune response in Aedes aegypti, modulating arbovirus infection of mosquitoes. Sindbis virus (SINV; family: Togaviridae, genus: Alphavirus) is an arbovirus that infects Ae. aegypti in the laboratory. SINV strain TR339 encounters a midgut escape barrier (MEB) during infection of Ae. aegypti. The nature of this barrier is not well understood. To investigate the role of the midgut as the central organ determining vector competence for arboviruses, we generated transgenic mosquitoes in which the RNAi pathway was impaired in midgut tissue of bloodfed females. We used these mosquitoes to reveal effects of RNAi impairment in the midgut on SINV replication, midgut infection and dissemination efficiencies, and mosquito longevity. Results As a novel tool for studying arbovirus-mosquito interactions, we engineered a transgenic mosquito line with an impaired RNAi pathway in the midgut of bloodfed females by silencing expression of the Aa-dcr2 gene. In midgut tissue of the transgenic Carb/dcr16 line, Aa-dcr2 expression was reduced ~50% between 1-7 days post-bloodmeal (pbm) when compared to the recipient mosquito strain. After infection with SINV-TR339EGFP, Aa-dcr2 expression levels were enhanced in both mosquito strains. In the RNAi pathway impaired mosquito strain SINV titers and midgut infection rates were significantly higher at 7 days pbm. There was also a strong tendency for increased virus dissemination rates among the transgenic mosquitoes. Between 7-14 days pbm, SINV was diminished in midgut tissue of the transgenic mosquitoes. Transgenic impairment of the RNAi pathway and/or SINV infection did not affect longevity of the mosquitoes. Conclusions We showed that RNAi impaired transgenic mosquitoes are a useful tool for studying arbovirus-mosquito interactions at the molecular level. Following ingestion by Ae. aegypti, the recombinant SINV-TR339EGFP was confronted with both MEB and a midgut infection barrier (MIB). Impairment of the RNAi pathway in the midgut strongly reduced both midgut barriers for the virus. This confirms that the endogenous RNAi pathway of Ae. aegypti modulates vector competence for SINV in the midgut. The RNAi pathway acts as a gatekeeper to the incoming virus by affecting infection rate of the midgut, intensity of infection, and dissemination from the midgut to secondary tissues.

Purpose: To determine the impact of long-term storage on adhesion between titanium and zirconia using resin cements. Materials and Methods: Titanium grade 4 blocks were adhesively fixed onto zirconia disks with four resin cements: Panavia F 2.0 (Kuraray Europe), GC G-Cem (GC Europe), RelyX Unicem (3M ESPE), and SmartCem 2 (Dentsply DeguDent). Shear bond strength was determined after storage in a water bath for 24 h, 16, 90, and 150 days at 37°C, and after 6000 cycles between 5°C and 55°C. Fracture behavior was evaluated using scanning electron microscopy. Results: After storage for at least 90 days and after thermocycling, GC G-Cem (16.9 MPa and 15.1 MPa, respectively) and RelyX Unicem (10.8 MPa and 15.7 MPa, respectively) achieved higher shear bond strength compared to SmartCem 2 (7.1 MPa and 4.0 MPa, respectively) and Panavia F2 (4.1 MPa and 7.4 MPa, respectively). At day 150, GC G-Cem and RelyX Unicem caused exclusively mixed fractures. SmartCem 2 and Panavia F2 showed adhesive fractures in one-third of the cases; all other fractures were of mixed type. After 24 h (GC G-Cem: 26.0, RelyX Unicem: 20.5 MPa, SmartCem 2: 16.1 MPa, Panavia F2: 23.6 MPa) and 16 days (GC G-Cem: 12.8, RelyX Unicem: 14.2 MPa, SmartCem 2: 9.8 MPa, Panavia F2: 14.7 MPa) of storage, shear bond strength was similar among the four cements. Conclusion: Long-term storage and thermocycling differentially affects the bonding of resin cement between titanium and zirconia. [ABSTRACT FROM AUTHOR]

Purpose: A key homeostatic mechanism of gingival fibroblasts in tissue repair is maintained through their production of tissue inhibitors of matrix metalloproteinases (TIMP). The purpose of this investigation was to elucidate the influence of diode laser irradiation on the in vitro production of TIMP-1 by human gingival fibroblasts. Materials and Methods: A human gingival fibroblast cell line (HGFB) was irradiated with a diode laser in pulsed mode at 0.5, 1, and 1.5 W for 5 s five times each with 5-s pauses in between. On the same plates in different wells, cells were then irradiated using continuous wave diode laser at 0.5 W for 5 s five consecutive times, with 5- s pauses in between. The cells were incubated overnight (18 h), and in the supernatants the concentration of free TIMP-1 was measured using a TIPM-1 ELISA. Results: At all settings, diode laser irradiation significantly increased in vitro TIMP-1 secretion of human gingival fibroblasts (p ≤ 0.001), measured after 18 hours of incubation. The best results were obtained using continuous wave irradiation with 0.5 W for 5 s applied five consecutive times with 5-s intervals in between. TIMP-1 secretion increased by 42.2% compared to the control. Conclusion: Our findings show that diode laser irradiation of human gingival fibroblasts signficantly increases TIMP-1 secretion compared to control values. This could contribute to periodontal tissue healing. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effect of bleaching agents containing different concentrations of hydrogen peroxide (HP) on color-change and on enamel-surface in bovine teeth. Furthermore the influence on cell viability and proliferation was investigated. Two hundred and forty teeth were randomly assigned into four groups (home bleaching ≤6%, in-office bleaching ≤6%, in-office bleaching > 6% HP, and control group). Bleaching was performed after artificial staining and the bleached index (BI) as well as the whiteness index (WI D ) was measured at several time points. Chemical analysis for HP concentrations and the pH of the bleaching products was done. Furthermore, enamel surfaces of randomly selected specimens were analyzed using scanning electron microscopy (SEM) and cytotoxicity of the tested bleaching products was evaluated in vitro using dental pulp cells (DPCs) and L929 cells. A statistically significant whitening effect was observed in almost all products. As expected all investigated products resulted in decreased cell viability, however, with different values of LC50 (median lethal concentration). SEM analysis showed an analog of enamel alterations with decreasing pH, increasing exposure time, and increasing HP concentration. Bleaching agents containing a low HP concentration are considered to be effective and to have less damaging effects on enamel and tested cells. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate 4 veneer composites--Signum+EM, Gradia Enamel E3, Sinfony E3, and SR Adoro S1--in a standardized test at 2 different conditions. For each composite, 2 groups of specimens (added to cultures immediately after preparation [ie, fresh] or after 7 days of incubation in cell culture medium) were added to L-929 fibroblast cultures for 72 hours. All composites showed reduced cell numbers compared to glass controls. Fresh specimens of Signum+EM exhibited the least cytotoxicity, followed by Gradia Enamel E3 and Sinfony E3 and then SR Adoro S1. For specimens with 7-day incubation, 3 of the composites (Signum+EM, Sinfony E3, and Gradia Enamel E3) showed similarly low cytotoxicity, while cytotoxic results with SR Adoro S1 were significantly higher. With low in vitro toxicity values in conjunction with good mechanical properties, veneer composites appear to offer an interesting alternative for prosthetic rehabilitation. [ABSTRACT FROM AUTHOR]

Purpose: To determine the performance of a resin composite material specially developed for core buildups in comparison with conventional restorative materials. Methods: 90 roughened ceramic blocs were divided into three groups; one group (n=30) was used for the core build-up material (Gradia Core) and the other two groups (n=30, each) were used for two conventional restorative materials (Tetric EvoCeram, Compoglass F). After adhesive fixation, specimens of each material were subdivided in accordance with the storage conditions (thermocycling or water storage). Shear bond strength was measured and fracture behavior was analyzed. Results: Gradia Core presented significantly higher shear bond strength values than the conventional restorative material Tetric EvoCeram, both after 24 hours water storage as well as after thermocycling. Compoglass F did not show any statistically significant differences compared to the other materials, independent of the storage condition. Flowever, Compoglass F resulted in numerically higher shear bond values than Tetric EvoCeram, but lower shear bond values than Gradia Core. Within the same materials, no statistically significant differences occurred regarding the storage conditions. [ABSTRACT FROM AUTHOR]