Isolating and confirming the Mudr­inserted flanking sequences of maize.

MuDR exhibits the highest transposition activity and insertional mutagenesis frequency in Mutator (Mu) family. If we isolate the MuDR­insertion­specific flanking sequences (MuDRFs), it will be crucial for using Mu element­mediated mutants. The MuDR­TAIL­PCR system was constructed and optimized using a combination of MuDR­TIR­nested specific primers and 12 arbitrary degenerate (AD) primers, modified reaction system and procedure and mutant DNA templates of 87 genotypes from M2 or M2:3 families created by crossing the W22::Mu line (active MuDR donor parent) from the UniformMu population with the Zong31 (Z31) line (recipient parent). Here 129 different MuDRFs were acquired by MuDR­TAIL­PCR, accounting for 86.60 % of the total mutant­specific agarose gel bands. In addition, we confirmed the authenticity of the non­redundant flanking sequence amplifications. The amplified non­redundant flanking sequences accounted for 65.12 % of the total MuDRFs, and 88.00 % of the non­redundant MuDRFs were inserted inside the genes. These results show that the MuDR­TAIL­PCR system that we developed can be used for specifically isolating MuDRFs.

[Study of the association between interleukin-1 polymorphisms and genetic susceptibility of coal workers' pneumoconiosis and silicosis].

Objective: To evaluate the relationship between IL-1 gene polymorphisms and coal workers' pneumoconiosis and silicosis susceptibility. Methods: We searched published full-text from PubMed, Web of Science, CNKI, VIP and Wanfang to collect case-control study on IL-1 gene polymorphisms with coal workers' pneumoconiosis and silicosis susceptibility. Eight articles, including 10 case-control studies were included in our study. All analyses were performed using the Stata version 12.0 software. Results: The IL-1RA (+2018) TC or CC variant genotypes were associated with coal workers' pneumoconiosis and silicosis risk (OR=1.65, 95%CI: 1.11-2.46) . In further stratified analyses, the IL-1RA (+2018) TC or CC variant genotypes were associated with an increased silicosis risk (OR=2.07, 95%CI: 1.45-2.95) , which were also associated with increased coal workers' pneumoconiosis and silicosis risk in Caucasians (OR=1.74, 95%CI: 1.22-2.47) . No significant association between IL-1ß (+3953) , IL-1ß (-511) , IL-1α (+4845) and coal workers' pneumoconiosis and silicosis risk was found either in the overall study or in the stratified analysis. Conclusion: These findings suggested that IL-1RA (+2018) may modify coal workers' pneumoconiosis and silicosis susceptibility. Further replication studies with large sample sizes are warranted to re-evaluate the relationship between IL-1RA (+2018) and coal workers' pneumoconiosis and silicosis risk.

A novel extracellular glycosidase activity from Rhodotorula mucilaginosa: its application potential in wine aroma enhancement.

UNLABELLED: The aim of the work was to evaluate the application potential of a glycosidase extract of one indigenous non-Saccharomyces strain in wine aroma enhancement. The isolate was selected from a local winemaking region in China for its high ß-glucosidase level and was identified as Rhodotorula mucilaginosa. The tolerance of the glycosidase extract to the typical winemaking conditions was assessed using the activity of its ß-glucosidase. After that, the hydrolysis capacity of R. mucilaginosa glycosidase for liberation of grape aroma glycosides was characterized in comparison to commercial enzyme preparations. Results of this work revealed that glycosidase extract from R. mucilaginosa proved to be active in the presence of 0-20% (w/v) glucose, 0-20% (v/v) ethanol and at pH 3·0-5·0. In the hydrolysis of aroma precursors, enzymes obtained from different origins possessed various levels of specificity and activity, showing high origin dependence (α = 0·05). Compared to commercial enzymes, the indigenous R. mucilaginosa glycosidase extract presented better catalytic preference for the 'fruity and floral' glycosides of benzenic compounds and C13 -norisoprenoids, but less sensitivity to the glycosides of C6 compounds and volatile phenols. SIGNIFICANCE AND IMPACT OF THE STUDY: This work presents a novel extracellular glycosidase preparation from an indigenous Rhodotorula mucilaginosa strain selected from a local winemaking region in China. This enzyme extract exhibits strong tolerance towards winemaking conditions. It shows hydrolysis specificity for glycosides of benzenic compounds and C13 -norisoprenoids, proving a potential candidate for improving floral and fruity aroma characteristics of wine.

beta-Catenin associates with the actin-bundling protein fascin in a noncadherin complex.

Catenins were first characterized as linking the cytoplasmic domains of cadherin cell-cell adhesion molecules to the cortical actin cytoskeleton. In addition to their essential role in modulating cadherin adhesivity, catenins have more recently been indicated to participate in cell and developmental signaling pathways. beta-Catenin, for example, associates directly with at least two receptor tyrosine kinases and transduces developmental signals within the Wnt pathway. Catenins also complex with the tumor suppressor protein adenomatous polyposis coli (APC), which appears to have a role in regulating cell proliferation. We have used the yeast two-hybrid method to reveal that fascin, a bundler of actin filaments, binds to beta-catenin's central Armadillo repeat domain. Western blotting of immunoprecipitates from cell line and mouse and rat brain extracts indicate that this interaction exists in vivo. Fascin and beta-catenin's association was further substantiated in vitro using purified proteins isolated from recombinant bacterial and baculoviral sources. Immunoprecipitation analysis indicates that fascin additionally binds to plakoglobin, which is highly homologous to beta-catenin but not to p120cas, a newly described catenin which contains a more divergent Armadillo-repeat domain. Immunoprecipitation, in vitro competition, and domain-mapping experiments demonstrate that fascin and E-cadherin utilize a similar binding site within beta-catenin, such that they form mutually exclusive complexes with beta-catenin. Immunofluorescence microscopy reveals that fascin and beta-catenin colocalize at cell-cell borders and dynamic cell leading edges of epithelial and endothelial cells. In addition to cell-cell borders, cadherins were unexpectedly observed to colocalize with fascin and beta-catenin at cell leading edges. It is conceivable that beta-catenin participates in modulating cytoskeletal dynamics in association with the microfilament-bundling protein fascin, perhaps in a coordinate manner with its functions in cadherin and APC complexes.