A distinct first replication cycle of DNA introduced in mammalian cells

Many mutation events in microsatellite DNA sequences were traced to the first embryonic divisions. It was not known what makes the first replication cycles of embryonic DNA different from subsequent replication cycles. Here we demonstrate that an unusual replication mode is involved in the first cycle of replication of DNA introduced in mammalian cells. This alternative replication starts at random positions, and occurs before the chromatin is fully assembled. It is detected in various cell lines and primary cells. The presence of single-stranded regions increases the efficiency of this alternative replication mode. The alternative replication cannot progress through the A/T-rich FRA16B fragile site, while the regular replication mode is not affected by it. A/T-rich microsatellites are associated with the majority of chromosomal breakpoints in cancer. We suggest that the alternative replication mode may be initiated at the regions with immature chromatin structure in embryonic and cancer cells resulting in increased genomic instability. This work demonstrates, for the first time, differences in the replication progression during the first and subsequent replication cycles in mammalian cells

Development and validation of a fluorimetric method to determine curcumin in lipid and polymeric nanocapsule suspensions

A simple, rapid, and sensitive fluorimetric method was developed and validated to quantify curcumin in lipid and polymeric nanocapsule suspensions, using acetonitrile as a solvent. The excitation and emission wavelengths were set at 397 nm and 508 nm, respectively. The calibration graph was linear from 0.1 to 0.6 µg/mL with a correlation coefficient of 0.9982. The detection and quantitation limits were 0.03 and 0.10 µg/mL, respectively. The validation results confirmed that the developed method is specific, linear, accurate, and precise for its intended use. The current method was successfully applied to the evaluation of curcumin content in lipid and polymeric nanocapsule suspensions during the early stage of formulation development.Um método fluorimétrico simples, rápido e sensível foi desenvolvido e validado para quantificação da curcumina em suspensões de nanocápsulas lipídicas e poliméricas, usando acetonitrila como solvente. Os comprimentos de onda de excitação e emissão foram 397 nm e 508 nm, respectivamente. Nas condições testadas, a curva de calibração demonstrou-se linear na faixa de 0,1 a 0,6 µg/mL, exibindo coeficiente de correlação de 0,9982. Os limites de detecção e quantificação foram 0,03 e 0,10 µg/mL, respectivamente. Os resultados da validação confirmaram que o método desenvolvido é específico, linear, exato e preciso para o uso proposto. O presente método foi aplicado com sucesso para a avaliação do teor de curcumina nas suspensões de nanocápsulas lipídicas e poliméricas durante o estágio inicial do desenvolvimento da formulação