RiboGreen is used for concentration measurements of RNA. Upon binding to the RNA, an approximately 1000-fold increase in sensitivity in comparison with the UV absorbance of the free polynucleotide is observed. In the present work, we demonstrate that this dye can penetrate in a time- and temperature-dependent manner the intact viral capsids of human rhinovirus serotypes 2 and 14, where it forms a fluorescent complex with the viral RNA. Capillary electrophoresis with laser-induced fluorescence detection of virus incubated with RiboGreen shows that the electrophoretic mobility of the viruses remained unchanged upon dye-binding. As shown for human rhinovirus serotype 2, its native conformation was conserved, since it still bound a recombinant soluble receptor fragment derived from the very low density lipoprotein receptor. The labeled RNA was released by heat-induced uncoating of the virus, and the RNA-dye complex could be directly detected if degradation was prevented with an RNase inhibitor. This in vitro labeling of viral RNA encased within a protein shell demonstrates the virion's dynamic nature that temporarily allows access of a low-molecular-mass compound to the otherwise protected RNA. It might be of great value for experiments requiring fluorescent viral particles with an unmodified surface, such as investigations of endocytosis and viral uncoating on the single molecule level.

Members of the low-density lipoprotein receptor family possess various numbers of ligand binding repeats that nonequally contribute to binding of minor group human rhinoviruses. Using an artificial concatemer of five copies of repeat 3 of the human very-low density lipoprotein receptor, we demonstrate protection of HRV2 against low-pH mediated uncoating and inhibition of penetration of an RNA-specific fluorescent dye into the intact virion. This indicates that the recombinant receptor inhibits viral breathing and irreversible conformational modifications of the capsid that precede RNA release, providing a new mechanism for rhinovirus neutralization by soluble receptor molecules.

Mesenchymal stem (stromal) cells (MSCs) possess unique biological characteristics such as plasticity, long term self-renewal, secretion of various bioactive molecules and ability of active migration to the diseased tissues that make them unique tool for regenerative medicine, nowadays. Until now MSCs were successfully derived from many tissue sources including bone marrow, umbilical cord, adipose tissue, dental pulp etc. The crucial step prior to their in vitro expansion, banking or potential clinical application is their separation. This review article aims to briefly describe the main MSCs separations techniques currently available, their basic principles, as well as their advantages and limits. In addition the attention is paid to the markers presently applicable for immunoaffinity-based separation of MSCs from different tissues and organs.

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