Amyotrophic Lateral Sclerosis-Associated Mutants of SOD1 Perturb mRNA Splicing through Aberrant Interactions with SRSF2.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder that results in the degeneration of neurons in the brain and spinal cord. Although a substantial number of studies have been conducted, much remains to be learned about the cellular mechanisms underlying ALS. In this study, we employed an engineered ascorbate peroxidase (APEX)-based proximity biotinylation, together with affinity pull-down of the ensuing biotinylated peptides, to investigate the proximity proteomes of human SOD1 and its two ALS-linked mutants, G85R and G93A. We were able to identify 25 common biotinylated peptides with preferential enrichment in the proximity proteomes of SOD1G85R and SOD1G93A over wild-type SOD1. Our coimmunoprecipitation followed by Western blot analyses revealed that one of these proteins, SRSF2, binds more strongly with the two SOD1 mutants than its wild-type counterpart. We also observed aberrant splicing of mRNAs in cells with ectopic expression of the two SOD1 mutants relative to cells expressing the wild-type protein. In addition, the aberrations in splicing elicited by the SOD1 variants were markedly attenuated upon knockdown of SRSF2. Collectively, we uncovered that ALS-liked SOD1G85R and SOD1G93A mutants interact more strongly with SRSF2, where the aberrant interactions perturbed mRNA splicing. Thus, our work offered novel mechanistic insights into the contributions of the ALS-linked SOD1 mutants to disease etiology.

Quality evaluation of chicken soup based on entropy weight method and grey correlation degree method.

This study aimed to develop an assessment framework for evaluating the quality of different chicken soup variants. Three types of chicken soup, traditional chicken soup (TCS), concentrated chicken soup (CCS), and blended chicken soup (BCS), were prepared and analyzed for various physicochemical parameters, including gross protein content, crude fat content, pH level, solid content, viscosity, and chromatic aberration value. Sensory evaluation was also conducted to assess overall quality. Correlation analysis helped identify three key evaluation indicators: gross protein content, L* value (lightness), and b* value (chromatic aberration). The weight assigned to gross protein content was the highest using the entropy weight method (EWM). Moreover, the grey correlation degree method was comprehensively applied to evaluate the chicken soup's quality. This analysis identified TCS and CCS as varieties with superior overall quality, showing a positive correlation with sensory evaluation, consistent with the results of nuclear magnetic resonance (NMR) used in this paper. These results provide theoretical support for assessing comprehensive quality and selecting chicken soup varieties.

Effects of different amounts of okara on texture, digestive properties, and microstructure of noodles.

As a byproduct of manufacturing soybeans, okara is high in dietary fiber, protein, and fats, and it contains all of the essential amino acids. Wheat, the primary ingredient in noodles, will lose nutrients during manufacturing, creating an imbalance in nutrients. This experiment is for the purpose of studying the effects of okara on quality, antioxidant properties, amino acid content, resistant starch (RS) content, and microstructure of noodles. The results indicate that the addition of 9% okara noodles increased hardness and adhesiveness by 107.19% and 132.14%, respectively, and improved ABTS free radical scavenging activity by 60.78%. The addition of 12% okara noodles increased the DPPH free radical scavenging ability by 23.66%, reduced the rapidly digestible starch (RDS) content of the noodles to 21.21%, and the resistant starch content increased to 44.85% (p < .05). Therefore, to address the issue of nutritional imbalance in wheat noodles without compromising the quality of the noodles, it is recommended to add 9% or 12% okara for the preparation of nutritionally fortified noodles.

MicroRNA­199a­5p regulates FOXC2 to control human vascular smooth muscle cell phenotypic switch.

Varicose veins are among the most common disorders of the vascular system; however, the pathogenesis of varicose veins remains unclear. The present study aimed to investigate the roles of microRNA (miR)­199a­5p in varicose veins and in the phenotypic transition of vascular smooth muscle cells (VSMCs). Bioinformatics analysis confirmed that miR­199a­5p had target sites on the forkhead box C2 (FOXC2) 3'­untranslated region. Reverse transcription­quantitative PCR (RT­qPCR) and western blotting were used to detect the expression levels of miR­199a­5p and FOXC2 in varicose vein and normal great saphenous vein tissues. Cell Counting Kit­8 and Transwell migration assays were performed to validate the effects of miR­199a­5p on VSMCs. Contractile markers, such as smooth muscle 22α, calponin, smooth muscle actin and myosin heavy chain 11 were used to detect phenotypic transition. RT­qPCR revealed that miR­199a­5p was downregulated in varicose veins compared with expression in normal great saphenous veins, whereas FOXC2 was upregulated in varicose veins. In addition, biomarkers of the VSMC contractile phenotype were downregulated in varicose veins. Overexpression of miR­199a­5p by mimics suppressed VSMC proliferation and migration, whereas depletion of miR­199a­5p enhanced VSMC proliferation and migration. Notably, the effects caused by miR­199a­5p could be reversed by FOXC2 overexpression. Dual luciferase reporter analysis confirmed that FOXC2 was a target of miR­199a­5p. In conclusion, miR­199a­5p may be a novel regulator of phenotypic switching in VSMCs by targeting FOXC2 during varicose vein formation.