ABSTRACT
The understanding of sex determination and differentiation in animals has recently made remarkable strides through the use of advanced research tools. At the gene level, the Mab-3-related transcription factor (Dmrt) gene family, which encodes for the typical DNA-binding doublesex/Mab-3 (DM) domain in their protein, is known for its contribution to sex determination and differentiation in insects. In this study, DNA-binding DM domain screening has identified eight transcripts from Artemia franciscana transcriptomic that encode proteins containing one conserved DNA-binding DM domain. The genome mapping confirmed that these eight transcripts are transcribed from six different loci on the A. franciscana genome assembly. One of those loci, the Af.dsx-4 locus, is closely related to Doublesex, a gene belonging to the Dmrt gene family. This locus could be transcribed into three alternative transcripts, namely Af.dsx4, Af.dsxF and Af.dsxM. While Af.dsx4 and Af.dsxF could putatively be translated to form an identical Af.dsxF protein of 186 aa long, Af.dsxM translates for an Af.dsxM protein of 289 aa long but shares a DNA-binding DM domain. Interestingly, Af.dsxF and Af.dsxM are confirmed as sex-specific transcripts, Af.dsxF is only present in females, and Af.dsxM is only present in male individuals. The results suggest that the sex-specific splicing mechanism of the doublesex described in insects is also present in A. franciscana. Af.dxs-4 locus can be used in further studies to clarify the sex determination pathways in A. fracnciscana.
Subject(s)
Alternative Splicing , Artemia , Animals , Female , Male , Artemia/genetics , RNA Splicing , DNAABSTRACT
OBJECTIVE Currently discussing the clinical treatment of pulmonary fibrosis commonly used drugs astragalus main active ingredient astragalosideⅣ(ASTⅣ)in vitro after transforming growth factor-β1 induced lung adenocarcinoma A549 epithelial cells after epithelial cell interstitial Epithelial-Mesenchymal Transition(EMT). METHODS The effect of astragalosideIV on the proliferation of A549 cells was detected by MTT assay for the first time.No significant effect of astragaloside on the prolifera-tion of A549 cells was found in the range of 1.25-20 μmol/L. A549 cells in vitro were divided into 5 groups: normal group, control group, low, medium and high experimental groups, which were treated for 72 hours,and the morphological changes of cells in each group were observed by light microscope. Real-time quantitative PCR (qPCR) and Western blotting were performed. Detection of gene and protein expression levels. RESULTS The results of real-time fluorescence quantitative PCR showed that the quantitative analysis of high-dose astragalosideⅣin the experimental group was lower than that of the control group in the α-SMA analysis,and the difference was statistically significant(P<0.05).The dose of AstragalosideⅣ in the experimental group was higher than that of the control group in the E-Cad analysis,and the difference was statistically significant(P<0.05).Western Blot results showed that the expression of α-SMA antibody in the high-dose and low-dose experimental group was lower than that in the control group, the difference was statistically significant (P<0.05). The high-dose experimental group had a significantly higher expression of E-Cad antibody than the control group, the difference was statistically significant (P<0.01). CONCLUSION This study uses A549 epithelial cells as a model, A549 cells were modeled and confirmed that Astragaloside can effectively inhibit TGF-β1-induced epithelial-mesenchymal transition(EMT)and provide a new basis for the treatment of pulmonary fibrosis.
ABSTRACT
Changes in the river flood regime may be due to atmospheric processes (e.g., increasing precipitation), catchment processes (e.g., soil compaction associated with land use change), and river system processes (e.g., loss of retention volume in the floodplains). This paper proposes a new framework for attributing flood changes to these drivers based on a regional analysis. We exploit the scaling characteristics (i.e., fingerprints) with catchment area of the effects of the drivers on flood changes. The estimation of their relative contributions is framed in Bayesian terms. Analysis of a synthetic, controlled case suggests that the accuracy of the regional attribution increases with increasing number of sites and record lengths, decreases with increasing regional heterogeneity, increases with increasing difference of the scaling fingerprints, and decreases with an increase of their prior uncertainty. The applicability of the framework is illustrated for a case study set in Austria, where positive flood trends have been observed at many sites in the past decades. The individual scaling fingerprints related to the atmospheric, catchment, and river system processes are estimated from rainfall data and simple hydrological modeling. Although the distributions of the contributions are rather wide, the attribution identifies precipitation change as the main driver of flood change in the study region. Overall, it is suggested that the extension from local attribution to a regional framework, including multiple drivers and explicit estimation of uncertainty, could constitute a similar shift in flood change attribution as the extension from local to regional flood frequency analysis.
ABSTRACT
A xylanase gene, xyn1, which encodes Paenibacillus sp. strain W-61 xylanase 1 (Xyn1), was cloned in Escherichia coli. xyn1 encodes 211 amino acid residues, including 28 amino acid residues of a signal peptide. The deduced amino acid sequence of the mature Xyn1 showed 95.7%, 84.0%, and 83.7% identity to family 11 xylanases of Aeromonas caviae ME-1, Paenibacillus sp., and Bacillus stearothermophilus respectively. The physico-chemical properties of recombinant Xyn1 were very similar to those of intact Xyn1, except for the molecular mass. The pattern of xylooligosaccharides generated by rXyn1 was investigated by fluorophore-assisted carbohydrate electrophoresis (FACE). The degradation rate of xylohexaose by rXyn1 increased markedly as compared with that of xylopentaose. Xylohexaose had a single preferential point of cleavage by rXyn1. On the basis of the pattern of action of xylooligosaccharides, the number of subsites was estimated to be six. The catalytic site was located between the third and the fourth subsites from non-reducing end.
