RESUMO
Polyrotaxane (PR) is a necklace-like supramolecule composed of cyclic components, such as cyclodextrin (CD), and a threading polymer capped with bulky end groups. PR exhibits peculiar mechanical properties attributed to the intermolecular cross-links with CD. Various CD molecules threaded on a linear PEG chain are often modified with chemical groups to add specific physicochemical properties. In general, the stoichiometry between CD and the PEG chain is a significant parameter that defines the unique physical properties of CD-based polyrotaxane (CD-PR). To date, mass spectrometry (MS) has been applied to investigate the molecular distribution of CD-PR, modifications of CD, and the threaded ratio of CD. However, only molecular weights (MWs) up to several 10s of kDa can be subjected to such analysis, whereas the MW of CD-PR used as industrial materials is much greater. Herein, we applied two ionic liquid matrices composed of 3-aminoquinoline and a high mass detector to analyze PRs using MALDI-TOF-MS. High to very high MW PRs in the range of 90-700 kDa were successfully analyzed using this method. The threaded ratio of CD was estimated from a single MW of CD, PEG, and PR. The ratios obtained were consistent with that obtained using 1H NMR. Furthermore, a single-stranded form of PR in γ-cyclodextrin threaded PR (γCD-PR) was clearly distinguished from a double-stranded form, which is only possible in γCD -PR because of its large host cavity.
RESUMO
To develop a new live attenuated mumps vaccine, a wild mumps Y7 strain isolated from a patient who developed mild parotitis was treated with nitrosoguanidine and ultraviolet, followed by selection of a temperature-sensitive clone. The selected clone, Y125, showed stable temperature-sensitivity in Vero cells. Intraspinal inoculation of marmosets with the Y125 produced only minimal histopathological changes, while intracerebral inoculation of neonatal rats revealed that the Y125 did not cause hydrocephalus. Both these effects of the Y125 were similar to those of the non-neurovirulent Jeryl Lynn strain. Furthermore, subcutaneous inoculation of the Y125 induced high levels of neutralizing antibodies in all Cercopithecus monkeys examined. Although the safety and immunogenicity should be confirmed in further field trials in humans, the present results indicate that the Y125 could be a promising vaccine candidate.