Assessing impacts of typhoons and the Chi-Chi earthquake on Chenyulan watershed landscape pattern in Central Taiwan using landscape metrics

The Chi-Chi earthquake (M-L = 7.3) occurred in the central part of Taiwan on September 21, 1999. After the earthquake, typhoons Xangsane and Toraji produced heavy rainfall that fell across the eastern and central parts of Taiwan on November 2000 and July 2001. This study uses remote sensing data, landscape metrics, multivariate statistical analysis, and spatial autocorrelation to assess how earthquake and typhoons affect landscape patterns. It addresses variations of the Chenyulan watershed in Nantou County, near the earthquake's epicenter and crossed by Typhoon Toraji. The subsequent disturbances have gradually changed landscape of the Chenyulan watershed. Disturbances of various types, sizes, and intensities, following various tracks, have various effects on the landscape patterns and variations of the Chenyulan watershed. The landscape metrics that are obtained by multivariate statistical analyses showed that the disturbances produced variously fragmented patches, interspersed with other patches and isolated from patches of the same type across the entire Chenyulan watershed. The disturbances also affected the isolation, size, and shape-complexity of patches at the landscape and class levels. The disturbances at the class level more strongly affected spatial variations in the landscape as well as patterns of grasslands and bare land, than variations in the watershed farmland and forest. Moreover, the earthquake with high magnitude was a starter to create these landscape variations in space in the Chenyulan watershed. The cumulative impacts of the disturbances on the watershed landscape pattern had existed, especially landslides and grassland in the study area, but were not always evident in space and time in landscape and other class levels

The Influence of Age and Sex on Genetic Associations with Adult Body Size and Shape : A Large-Scale Genome-Wide Interaction Study

Genome-wide association studies (GWAS) have identified more than 100 genetic variants contributing to BMI, a measure of body size, or waist-to-hip ratio (adjusted for BMI, WHRadjBMI), a measure of body shape. Body size and shape change as people grow older and these changes differ substantially between men and women. To systematically screen for age-and/or sex-specific effects of genetic variants on BMI and WHRadjBMI, we performed meta-analyses of 114 studies (up to 320,485 individuals of European descent) with genome-wide chip and/or Metabochip data by the Genetic Investigation of Anthropometric Traits (GIANT) Consortium. Each study tested the association of up to similar to 2.8M SNPs with BMI and WHRadjBMI in four strata (men 50y, women 50y) and summary statistics were combined in stratum-specific meta-analyses. We then screened for variants that showed age-specific effects (G x AGE), sex-specific effects (G x SEX) or age-specific effects that differed between men and women (G x AGE x SEX). For BMI, we identified 15 loci (11 previously established for main effects, four novel) that showed significant (FDR= 50y). No sex-dependent effects were identified for BMI. For WHRadjBMI, we identified 44 loci (27 previously established for main effects, 17 novel) with sex-specific effects, of which 28 showed larger effects in women than in men, five showed larger effects in men than in women, and 11 showed opposite effects between sexes. No age-dependent effects were identified for WHRadjBMI. This is the first genome-wide interaction meta-analysis to report convincing evidence of age-dependent genetic effects on BMI. In addition, we confirm the sex-specificity of genetic effects on WHRadjBMI. These results may providefurther insights into the biology that underlies weight change with age or the sexually dimorphism of body shape.Peer reviewe

Secreted Production of Renilla Luciferase in Bacillus subtilis

Luciferase (Rluc) from the soft coral Renilla reniformis has been widely used as a bioluminescent reporter, and its secreted production has been solely performed in mammalian cells thus far. To make the production more efficient, a series of approaches was attempted to overproduce Rluc extracellularly in Bacillus subtilis. First, Cys124 in the Rluc gene was substituted with Ala. The mutant gene was subsequently incorporated into a pUB110/R6K-based plasmid, consequently, fusing with the P43 promoter and the sacB signal peptide. With the nitrogen-rich medium, B. subtilis strain bearing the plasmid became able to secret a detectable amount of Rluc. Moreover, the secretion signal for the Rluc gene was replaced by the aprN leader peptide with or without the propeptide. The result led to a more than twofold increase in the secreted Rluc. Finally, by enhancing the transcription of the Rluc gene implementing the P43 and spac tandem promoter, it resulted in the secreted Rluc with a yield of 100 mg/L. Overall, this study illustrates a potential strategy for improving the secretion efficiency of heterologous proteins in B. subtilis. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 26: 589-594, 201

Medium optimization for the production of recombinant nattokinase by Bacillus subtilis using response surface methodology

Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases. Most recently, a new Bacillus subtilis/Escherichia coli (B. subtilis/E. coli) shuttle vector has been developed to achieve stable production of recombinant nattokinase in B. subtilis (Chen; et al. 2007, 23, 808-813). With this developed B. subtilis strain, the design of an optimum but cost-effective medium for high-level production of recombinant nattokinase was attempted by using response surface methodology. On the basis of the Plackett-Burman design, three critical medium components were selected. Subsequently, the optimum combination of selected factors was investigated by the Box-Behnken design. As a result, it gave the predicted maximum production of recombinant nattokinase with 71 500 CU/mL for shake-flask cultures when the concentrations of soybean hydrolysate, potassium phosphate, and calcium chloride in medium were at 6.100, 0.415, and 0.015%, respectively. This was further verified by a duplicated experiment. Moreover, the production scheme based on the optimum medium was scaled up in a fermenter. The batch fermentation of 3 L was carried out by controlling the condition at 37 degrees C and dissolved oxygen reaching 20% of air saturation level while the fermentation pH was initially set at 8.5. Without the need for controlling the broth pH, recombinant nattokinase production with a yield of 77 400 CU/mL (corresponding to 560 mg/L) could be obtained in the culture broth within 24 h. In particular, the recombinant B. subtilis strain was found fully stable at the end of fermentation when grown on the optimum medium. Overall, it indicates the success of this experimental design approach in formulating a simple and cost-effective medium, which provides the developed strain with sufficient nutrient supplements for stable and high-level production of recombinant, nattokinase in a fermenter

Strategy to approach stable production of recombinant nattokinase in Bacillus subtilis

Bacillus subtilis (B. subtilis) is widely accepted as an excellent host cell for the secretory production of recombinant proteins. In this study, a shuttle vector was constructed by fusion of Staphylococcus aureus (S. aureus) plasmid pUB110 with Escherichia coli (E. coli) plasmid pUC18 and used for the expression of nattokinase in B. subtilis. The pUB110/pUC-based plasmid was found to exhibit high structural instability with the identification of a DNA deletion between two repeated regions. An initial attempt was made to eliminate the homologous site in the plasmid, whereas the stability of the resulting plasmid was not improved. In an alternative way, the pUC18-derived region in this hybrid vector was replaced by the suicidal R6K plasmid origin of E. coli. As a consequence, the pUB110/R6K-based plasmid displayed full structural stability, leading to a high-level production of recombinant nattokinase in the culture broth. This was mirrored by the detection of a very low level of high molecular weight DNAs generated by the plasmid. Moreover, 2-fold higher nattokinase production was obtained by B. subtilis strain carrying the pUB110/R6K-based plasmid as compared to the cell with the pAM beta 1-derived vector, a plasmid known to have high structural stability. Overall, it indicates the feasibility of the approach by fusing two compatible plasmid origins for stable and efficient production of recombinant nattokinase in B. subtilis

Medium optimization and production of secreted Renilla luciferase in Bacillus subtilis by fed-batch fermentation

Luciferase of the soft coral Renilla reniformis (Rluc) has found a multitude of potential applications. However, its production in a secreted form has been solely performed in mammalian cells. In this work, a high production of secreted Rluc was approached in Bacillus subtilis by implementing the fermentation strategy. First, three critical medium components among others were selected by the Plackett-Burman design. Followed by the Box-Behnken design, the medium formulation was optimized and composed of 1.17% glucose, 2.27% yeast extract, and 0.55% glutamate. With the optimum medium, the shake-flask culture of the producer strain was carried out to secret Rluc with the yield of 55 mg/L. Finally, the production scheme was scaled up using a 5-L jar fermenter operated in a fed-batch mode. By controlling the operational condition, the feeding solution was pumped in an impulse mode to the culture. As a consequence, the production of Rluc reaching 500 mg/L was obtained after fermentation for 27 h. (C) 2010 Elsevier B.V. All rights reserved

Applicability of new expression vectors for both engineering uses and biological studies

To be applicable for both engineering and biological uses, the plasmid with the features of tight regulation, high-level expression, and subtle modulation (or homogeneous induction) is required. IPTG-inducible promoters are of particular interest since they acquire the latter two merits but usually lack stringency. To this end, two plamids have been developed to contain the T7 A1 promoter along with either lacI(q) or lacI gene. As a production system, the cells harboring the plamids with the lacZ gene clone enabled production of the maximal protein accounting for 35% total cell content upon induction by a saturating IPTG level. This protein yield is amplified over 700-fold relative to that at the uninduced state. As a system for biological study, the ppc negative strain bearing the plasmid with the ppc gene clone failed to grow on glucose without IPTG induction but immediately resumed its growth in the presence of IPTG. Moreover, the level of the ppc gene product in the cell was varied by various IPTG, and the result revealed that the wild-type ppc level was sufficient to support the saturated growth of the cell on glucose. Overall, it illustrates the applicability of these plasmids to needs in the post-genome era

Finite-difference frequency-domain analysis of 2-D photonic crystals with curved dielectric interfaces

An improved finite-difference frequency-domain method based on Taylor series expansion, local coordinate transformation, and boundary condition matching is developed for analyzing the band diagrams of 2-D photonic crystals. This newly proposed scheme can deal with piecewise homogeneous structures with curved dielectric interfaces. It also shows much better performance in modeling structures of highly index contrast ratio. We have verified this improved scheme with frequently-used plane-wave expansion method through the calculation of the band diagrams of a 2-D photonic crystal with square lattice. We also compare the scheme with the staircase approximation. We further improve the convergence behavior by adopting high-order terms. The result of this adopting is also demonstrated