No abstract

Inference of gene sequences in ancestral species has been widely used to test hypotheses concerning the process of molecular sequence evolution. However, the approach may produce spurious results, mainly because using the single best reconstruction while ignoring the suboptimal ones creates systematic biases. Here we implement methods to correct for such biases and use computer simulation to evaluate their performance when the substitution process is nonstationary. The methods we evaluated include parsimony and likelihood using the single best reconstruction (SBR), averaging over reconstructions weighted by the posterior probabilities (AWP), and a new method called expected Markov counting (EMC) that produces maximum-likelihood estimates of substitution counts for any branch under a nonstationary Markov model. We simulated base composition evolution on a phylogeny for six species, with different selective pressures on G+C content among lineages, and compared the counts of nucleotide substitutions recorded during simulation with the inference by different methods. We found that large systematic biases resulted from (i) the use of parsimony or likelihood with SBR, (ii) the use of a stationary model when the substitution process is nonstationary, and (iii) the use of the Hasegawa-Kishino-Yano (HKY) model, which is too simple to adequately describe the substitution process. The nonstationary general time reversible (GTR) model, used with AWP or EMC, accurately recovered the substitution counts, even in cases of complex parameter fluctuations. We discuss model complexity and the compromise between bias and variance and suggest that the new methods may be useful for studying complex patterns of nucleotide substitution in large genomic data sets.

A methanol extract from the rhizomes of Kaempferia parviflora Wall. ex Baker (Zingiberaceae) has shown inhibitory effects against melanogenesis in theophylline-stimulated murine B16 melanoma 4A5 cells (IC50 = 9.6 μg/mL). Among 25 flavonoids and three acetophenones isolated previously (1-28), several constituents including 5-hydroxy-7,3',4'-trimethoxyflavone (6, IC50 = 8.8 μM), 5,7,3',4'-tetramethoxyflavone (7, 8.6 μM), 5,3'-dihydroxy-3,7,4'-trimethoxyflavone (12, 2.9 μM), and 5-hydroxy-3,7,3',4'-tetramethoxyflavone (13, 3.5 μM) showed inhibitory effects without notable cytotoxicity at the effective concentrations. Compounds 6, 7, 12, and 13 inhibited the expression of tyrosinase, tyrosine-related protein (TRP)-1, and TRP-2 mRNA, which could be the mechanism of their melanogenesis inhibitory activity. In addition, a quantitative analytical method for 12 methoxyflavones (1, 2, 4-11, 13, and 14) in the extract was developed using HPLC. The optimal condition for separation and detection of these constituents were achieved on an ODS column (3 μm particle size, 2.1 mm i.d. × 100 mm) with MeOH-0.1 % aqueous acetic acid solvent systems as the mobile phase, and the detection and quantitation limits of the method were estimated to be 0.08-0.66 ng and 0.22-2.00ng, respectively. The relative standard deviation values of intra- and interday precision were lower than 0.95 and 1.08 %, respectively, overall mean recoveries of all flavonoids were 97.9-102.9 %, and the correlation coefficients of all the calibration curves showed good linearity within the test ranges. For validation of the protocol, extracts of three kinds of the plant's rhizomes collected from different regions in Thailand (Leoi, Phetchabun, and Chiang Mai provinces) were evaluated. The results indicated that the assay was reproducible, precise, and could be readily utilized for the quality evaluation of the plant materials.

The elasticity of the periodic bundle structure formed by the interaction between the tip of an atomic force microscope and a polycarbonate surface was studied. The local elasticity of the bundle was observed using ultrasonic force microscopy, which combines the sensitivity to an elastic structure of acoustic microscopy with atomic force microscopy. The mean height of the bundle increases with increase in the number of scan-scratching cycles. The process of decreasing elasticity of a growing bundle of polycarbonate was observed clearly. Furthermore, the elasticity contrast inside the bundle structure was observed by adjusting the amplitude of ultrasonic vibration of sample height in ultrasonic force microscopy.

No abstract

A growing number of molecular evolutionary studies are estimating the proportion of adaptive amino acid substitutions (α) from comparisons of ratios of polymorphic and fixed DNA mutations. Here, we examine how violations of two of the model assumptions, neutral evolution of synonymous mutations and stationary base composition, affect α estimation. We simulated the evolution of coding sequences assuming weak selection on synonymous codon usage bias and neutral protein evolution, α = 0. We show that weak selection on synonymous mutations can give polymorphism/divergence ratios that yield α-hat (estimated α) considerably larger than its true value. Nonstationary evolution (changes in population size, selection, or mutation) can exacerbate such biases or, in some scenarios, give biases in the opposite direction, α-hat < α. These results demonstrate that two factors that appear to be prevalent among taxa, weak selection on synonymous mutations and non-steady-state nucleotide composition, should be considered when estimating α. Estimates of the proportion of adaptive amino acid fixations from large-scale analyses of Drosophila melanogaster polymorphism and divergence data are positively correlated with codon usage bias. Such patterns are consistent with α-hat inflation from weak selection on synonymous mutations and/or mutational changes within the examined gene trees.

Polystyrene (PS) thin film modified with a novel ultrasonic scratching method has been studied using an atomic force microscope (AFM). Ultrasonic scratching can be realized by a combination of an AFM and a quartz crystal resonator (QCR). The PS thin film was deposited on the surface of an AT-cut QCR which oscillates with mechanical shear deformation at its resonant frequency (6.5 MHz) with amplitude of a few nm. The oscillating surface is scratched with the AFM’s diamond tip to generate an effective cutting force. Scratching without a QCR oscillation forms bumps on the PS surface, which is known well to be a unique phenomenon of scratched polymer surfaces. However, ultrasonic scratching can result in the carving of PS thin films without the formation of bumps. The depressed bottom carved by ultrasonic scratching is flat in comparison with that scratched without surface oscillations. Furthermore, on the previously modified area of ultrasonic scratching, bumps are not formed, even after overlapping of the scratching without surface oscillations.

Photoelectron holography has been expected to be a powerful tool for visualizing three-dimensional (3D) local atomic structures around a photoelectron emitter atom. Photoelectron emitter atom sites with different elements can be distinguished by photoelectron kinetic energy. The elements of atoms reconstructed in atomic images, however, could not be assigned. We developed a method for the elemental characterization of the reconstructed atoms using a small difference of the scattered electron waves and demonstrated it using a measured InP(001) photoelectron hologram. Element assignment for both the emitter atom and the reconstructed atoms in 3D atomic images became possible.