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HFF-1 cells were plated into six-well plates to a density of 10 5 cells/mL and allowed to incubate at 37C in a 5% (v/v) CO 2 under static conditions, with the fresh supplemented DMEM growth medium being added every alternative day until a confluence of 95%100% of cells (verified using a light microscope) were achieved

The conditioned media were collected and centrifuged at 500 g for 5 min to remove cell debris, filtered with a 0.45 m filter, and concentrated to ~100 l using 10 kDa cutoff Amicon Centrifugal filters (Millipore-Sigma)

Although not documented in Keshan disease, it is possible that selenium deficiency may increase the virulence of viruses with the potential to invade and damage the heart muscle (57)

It is preferable to test the product on a small area before full use, especially for sensitive skin

Powers, H

Oxidative stress induced damage to lipids, proteins and DNA result in neuronal death which leads to AD