RESUMO
Genome packaging of double-stranded RNA (dsRNA) phages has been widely studied using biochemical and molecular biology methods. We adapted the existing in vitro packaging system of one such phage for single-molecule experimentation. To our knowledge, this is the first attempt to study the details of viral RNA packaging using optical tweezers. Pseudomonas phage φ6 is a dsRNA virus with a tripartite genome. Positive-sense (+) single-stranded RNA (ssRNA) genome precursors are packaged into a preformed procapsid (PC), where negative strands are synthesized. We present single-molecule measurements of the viral ssRNA packaging by the φ6 PC. Our data show that packaging proceeds intermittently in slow and fast phases, which likely reflects differences in the unfolding of the RNA secondary structures of the ssRNA being packaged. Although the mean packaging velocity was relatively low (0.07-0.54 nm/sec), packaging could reach 4.62 nm/sec during the fast packaging phase.
Assuntos
Bacteriófago phi 6/fisiologia , RNA Viral/genética , Bacteriófago phi 6/genética , Técnicas In Vitro , Modelos Moleculares , Conformação de Ácido Nucleico , Dobramento de RNA , RNA Viral/química , Montagem de VírusRESUMO
Scratch resistance (SR) of externally plasticized hydroxypropyl methylcellulose (HPMC) films intended for tablet film coatings was studied. Special attention was paid to effects of short-term aging and ultraviolet (UV) treatment on the SR properties of these films. Controlled scratching of the films was performed with a Lloyd LRX materials tester featuring a spherical steel tip. Scratch surface profiles were measured by scanning white light interferometry (SWLI). The influence of using an external plasticizer on the SR was studied by comparing scratch dimensions in non-plasticized films to samples plasticized either with glycerol or polyethylene glycol (PEG) 400. The study demonstrates that both the amount and type of plasticizer influences the SR of aqueous HPMC films. It also shows that SWLI can quantitatively evaluate the effect of plasticizer content and aging on the SR of pharmaceutical films. This knowledge could be used to optimize pharmaceutical film coating formulations.
Assuntos
Metilcelulose/análogos & derivados , Plastificantes/química , Comprimidos/química , Tecnologia Farmacêutica/métodos , Glicerol/química , Dureza , Derivados da Hipromelose , Teste de Materiais/instrumentação , Teste de Materiais/métodos , Metilcelulose/química , Polietilenoglicóis/química , Fatores de Tempo , Raios UltravioletaRESUMO
A supercontinuum light source was incorporated into a custom-built scanning white-light interferometer. This light source based on a Nd:YAG pumped microstructured optical fiber exhibits 1.21+/-0.10 microm temporal coherence length. The device operation was validated by characterizing the step height on a microelectromechanical system component. The measured step height- 7.027+/-0.020 microm-agreed with results obtained by employing traditional light sources: a halogen lamp and a white light-emitting diode. The new light source features high output intensity of 20-35 mW, which is beneficial when measuring low-reflectivity samples. As the supercontinuum light source may be modulated at frequencies exceeding 10 MHz, it holds potential for stroboscopic dynamic measurements.
