Quercetin 7-rhamnoside reduces porcine epidemic diarrhea virus replication via independent pathway of viral induced reactive oxygen species.

BACKGROUND: On the base of our previous study we were observed relevant studies on the hypothesis that the antiviral activity of quercetin 7-rhamnoside (Q7R), a flavonoid, won't relate ability of its antioxidant. METHODS: We were investigated the effects of Q7R on the cytopathic effects (CPE) by CPE reduction assay. Production of DNA fragment and reactive oxygen species (ROS) induced by PEDV infection were studied using DNA fragmentation assay and flow cytometry. RESULTS: In the course of this study it was discovered that Q7R is an extremely potent compound against PEDV. The addition of Q7R to PEDV-infected Vero cells directly reduced the formation of a visible cytopathic effect (CPE). Also, Q7R did not induce DNA fragmentation. Furthermore, ROS increased the infection of PEDV, which was strongly decreased by N-acetyl-L-cysteins (NAC). However, the increased ROS was not decreased by Q7R. Antiviral activity of antioxidants such as NAC, pyrrolidine dithiocarbamate (PDTC), and the vitamin E derivative, trolox, were hardly noticed. CONCLUSIONS: We concluded that the inhibition of PEDV production by Q7R is not simply due to a general action as an antioxidants and is highly specific, as several other antioxidants (NAC, PDTC, trolox) are inactive against PEDV infection.

A scaffold-free surface culture of B16F10 murine melanoma cells based on magnetic levitation.

Multicellular spheroids are obtained in a variety of three-dimensional (3D) culture systems without the use of supporting scaffold. We present here a 3D culture method that resulted in a multicellular sheet under scaffold-free conditions. A floating disk-shaped 3D culture was prepared by magnetic levitation of B16F10 cells that has ingested Fe3O4-containing fibroin microspheres. The melanoma disk grew up to 19 mm in diameter and the thickness was ranged between 80 and 100 µm. The 3D culture was filled with closely packed cells that were proliferating exponentially at a specific growth rate of µ = 0.015 h-1. Approximately half of the cells were Ki-67 positive with no detectable levels of apoptotic or autophagic cells. However, the percentage of propidium iodide-permeable cells was 8.5 ± 1.2 %, which was probably due to physical damage in the cell membrane caused by Fe3O4-containing microspheres under a strong magnetic field. Melanin production increased by a factor of 3.0-3.7 in the 3D culture, due to an increased population of pigmented cells. This study presented a surface 3D culture of B16F10 cells without the use of a scaffold based on magnetic levitation.