Structural and functional analysis of four non-coding Y RNAs from Chinese hamster cells: identification, molecular dynamics simulations and DNA replication initiation assays

Abstract Background The genes coding for Y RNAs are evolutionarily conserved in vertebrates. These non-coding RNAs are essential for the initiation of chromosomal DNA replication in vertebrate cells. However thus far, no information is available about Y RNAs in Chinese hamster cells, which have already been used to detect replication origins and alternative DNA structures around these sites. Here, we report the gene sequences and predicted structural characteristics of the Chinese hamster Y RNAs, and analyze their ability to support the initiation of chromosomal DNA replication in vitro. Results We identified DNA sequences in the Chinese hamster genome of four Y RNAs (chY1, chY3, chY4 and chY5) with upstream promoter sequences, which are homologous to the four main types of vertebrate Y RNAs. The chY1, chY3 and chY5 genes were highly conserved with their vertebrate counterparts, whilst the chY4 gene showed a relatively high degree of diversification from the other vertebrate Y4 genes. Molecular dynamics simulations suggest that chY4 RNA is structurally stable despite its evolutionarily divergent predicted stem structure. Of the four Y RNA genes present in the hamster genome, we found that only the chY1 and chY3 RNA were strongly expressed in the Chinese hamster GMA32 cell line, while expression of the chY4 and chY5 RNA genes was five orders of magnitude lower, suggesting that they may in fact not be expressed. We synthesized all four chY RNAs and showed that any of these four could support the initiation of DNA replication in an established human cell-free system. Conclusions These data therefore establish that non-coding chY RNAs are stable structures and can substitute for human Y RNAs in a reconstituted cell-free DNA replication initiation system. The pattern of Y RNA expression and functionality is consistent with Y RNAs of other rodents, including mouse and rat

Evaluation of rare earth oxides doping SnO2.(Co1/4,Mn3/4)O-based varistor system

The present paper aims to verify the inuence of rare earth oxide such as lanthanum (La2O3) and neodymium (Nd2O3) doping SnO2 + 0.25%CoO + 0.75%MnO2 + 0.05%Ta2O5 system. The analysis focus on microstructural inuence on electrical properties. Microstructural analysis were made by using Transmission Electron Microscopy (TEM) at different regions of the samples. From such analysis it was found that La2O3 and Nd2O3 oxides cause heterogeneous segregation and precipitation at grain boundary concerning cobalt and manganese, decreasing the nonohmic electrical properties, as discussed, likely due to the increasing of grain boundary non-active potential barriers

Micro-sized erosions in a nanofilled composite after repeated acidic beverage exposures: consequences of clusters dislodgments

Objective: To evaluate the hardness (KHN), color stability (DE), and superficial micromorphology of two categories of composites after immersion in either distilled water or grape juice for up to 45 days. MATERIAL AND METHODS: Cylindrical specimens (6 mm diameter x 2 mm thick) were obtained according to the factors: composite [Opallis (FGM) and Filtek Z350XT (3M ESPE)]; immersion solution (distilled water and grape juice); and evaluation time: 24 h and 7, 14, 21, 28, and 45 days. After photoactivation, the specimens were stored at 37ºC for 24 h. KHN (50 g/15 s) and ΔE were then repeatedly assessed according to the immersion solutions. Data were analyzed (three-way ANOVA/Tukey's test). Scanning electron microscopy (SEM) topographic analysis was also performed. RESULTS: In general, KHN of both composites reduced after 24 h, irrespective of the immersion solution and time. A significantly lower KHN was noted for Opallis compared with Filtek Z350XT in all parameters. An increase in ΔE over time was noted for both composites, irrespective of the immersion solution. Significantly higher ΔE was noted for Filtek Z350XT immersed in grape juice compared with Opallis, regardless of the evaluation time. The grape juice caused significantly higher DE compared with water in all periods. SEM analysis showed eroded areas for Filtek Z350XT but not for Opallis. CONCLUSIONS: The compositions and immersion solutions influence the composite hardness and the color stability. In spite of the higher hardness, the nanofilled composite is more susceptible to color change than the microhybrid when immersed in an acidic dyed solution