Search

In this study, a magnetic solid-phase extraction (MSPE) method coupled with High-Performance Liquid Chromatography Mass Spectrometry (HPLC–MS/MS) for the determination of illegal basic dyes in food samples was developed and validated. This method was based on Magnetic sulfonated reduced graphene oxide (M-S-RGO), which was sensitive and selective to analytes with structure of multiaromatic rings and negatively charged ions. Several factors affecting MSPE efficiency such as pH and adsorption time were optimized. Under the optimum conditions, the calibration curves exhibited good linearity, ranging from 5 to 60 µg/g with correlation coefficients >0.9950. The limits of detection of 16 basic dyes were in the range of 0.01–0.2 µg/L. The recoveries ranged from 70% to 110% with RSD% < 10%. The results indicate that M-S-RGO is an efficient and selective adsorbent for the extraction and cleanup of basic dyes. Due to the MSPE procedures, matrix effect and interference were eliminated in the analysis of HPLC–MS/MS without the matrix-matched standards. Thus, validation data showed that the proposed MSPE–HPLC–MS/MS method was rapid, efficient, selective, and sensitive for the determination of illegal basic dyes in foods.

We assembled and simulated putative models including plexin A4 resepctively complexed with RND1 and RhoD, to investigate the structural mechanisms underlying RND1 as an activator and RhoD as an attenuator in plexin signaling. The models suggested that RND1 binding is compatible with the dimerization of plexin A4 while RhoD binding is likely disruptive to the dimerization. The models also showed two distinct modes of interactions of RND1 and RhoD with the membrane: RND1 interacts with the membrane loosely and its long C-terminal tail serves as a flexible tether to the membrane, whereas RhoD interacts with the membrane in a specific manner using a positively-charged membrane interface, which is absent in RND1. Importantly, with RhoD binding at the RBD, the buttress interaction with the dimerization helix in the plexin dimer is much reduced compared to that in the RND1-bound dimer, suggesting that RhoD weakens the buttress interaction with the dimerization helix and potentially destabilizes the plexin dimer. In summary, RhoD binding destabilizes the RBD with respect to the GAP domain, destabilizing the buttress segment with respect to the dimerization interface, and ultimately leads to destabilization of the dimer interface. In contrast, by the same RBD-centered route, RND1 binding helps stabilize the plexin dimer.

The Warburg effect (aerobic glycolysis) is a hallmark of cancer and is becoming a promising target for diagnosis and therapy. Phosphoglycerate kinase 1 (PGK1) is the first adenosine triphosphate (ATP)-generating glycolytic enzyme in the aerobic glycolysis pathway and plays an important role in cancer development and progression. However, how microRNAs (miRNAs) regulate PGK1-mediated aerobic glycolysis remains unknown. Here, we show that miR-16-1-3p inhibits PGK1 expression by directly targeting its 3'-untranslated region. Through inhibition of PGK1, miR-16-1-3p suppressed aerobic glycolysis by decreasing glucose uptake, lactate and ATP production, and extracellular acidification rate, and increasing oxygen consumption rate in breast cancer cells. Aerobic glycolysis regulated by the miR-16-1-3p/PGK1 axis is critical for modulating breast cancer cell proliferation, migration, invasion and metastasis