Fabry disease due to deficiency of α-galactosidase A (α-Gal) causes lysosomal accumulation of globotriaosylceramide (Gb3) in multiple tissues and prominently in the vascular endothelium. Although enzyme replacement therapy (ERT) by injection of recombinant α-Gal improves the disease outcome, effects on the vasculopathy associated to life-threatening cerebrovascular, cardiac and renal complications are still limited. We designed a strategy to enhance delivery of α-Gal to organs and endothelial cells (ECs). We targeted α-Gal to intercellular adhesion molecule 1 (ICAM-1), a protein expressed on ECs throughout the vasculature, by loading this enzyme on nanocarriers coated with anti-ICAM (anti-ICAM/α-Gal NCs). In vitro radioisotope tracing showed efficient loading of α-Gal on anti-ICAM NCs, stability of this formulation under storage and in model physiological fluids, and enzyme release in response to lysosome environmental conditions. In mice, delivery of 125 I-α-Gal was markedly enhanced by anti-ICAM/ 125 I-α-Gal NCs in brain, kidney, heart, liver, lung, and spleen, and transmission electron microscopy showed anti-ICAM/α-Gal NCs attached to and internalized into the vascular endothelium. Fluorescence microscopy proved targeting, endocytosis and lysosomal transport of anti-ICAM/α-Gal NCs in macro-and micro-vascular ECs, and a marked enhancement of Gb3 degradation. Therefore, ICAM-1-targeting strategy may help improve the efficacy of therapeutic enzymes for Fabry disease.

oxygen species; TLR, toll-like receptor; TRAF3, tumor necrosis factor receptorassociated factor 3; ULK1, Unc-51 like autophagy activating kinase 1. AbstractDisrupted mitochondrial function and reactive oxygen species (ROS) generation cause cellular damage and oxidative stress-induced macrophage inflammatory cell death. It remains unclear how mitochondrial dysfunction relates to inflammasome activation and pyroptotic cell death. In this study, we demonstrated that tumor necrosis factor receptor-associated factor 3 (TRAF3) regulates mitochondrial ROS production and promotes TLR agonist LPS plus nigericin (LPS/Ng)-induced inflammasome and pyroptosis in mouse primary macrophages and human monocyte THP-1 cells. Co-IP assays confirmed that TRAF3 forms a complex with TRAF2 and cIAP1 and mediates ubiquitin and degradation of Unc-51 like autophagy activating kinase 1 (ULK1). Moreover, knockdown of ULK1 in THP-1 cells significantly promoted LPS/Ng-induced inflammasome by activating caspase 1 and mature IL-1β. Apoptosis inducing factor (AIF) translocation from mitochondrial to nuclear was observed in ULK1-deficient THP-1 cells under LPS/Ng stimulation, which mediates LPS/Ng-induced cell death in ULK1 deficient macrophages. In conclusion, this study identified a novel role of TRAF3 in regulation of ULK1 ubiquitination and inflammasome signaling and provided molecular mechanisms by which ubiquitination of ULK1 controls mitochondrial ROS production, inflammasome activity, and AIF-dependent pyroptosis. K E Y W O R D S cell death, inflammasome, macrophages, mitochondrial, reactive oxygen species | 7145 SHEN Et al.

A reversed-phase simultaneous determination method for the enantiomers of carfentrazone-ethyl and its metabolite carfentrazone in agricultural and environmental samples was established using high-performance liquid chromatography-tandem mass spectrometry. A complete enantioseparation of carfentrazone-ethyl and its chiral metabolite carfentrazone enantiomers was obtained using a chiral column. The absolute configuration and specific optical rotation of carfentrazone-ethyl and carfentrazone enantiomers were first confirmed as peaks 1, 2, 3, and 4: S-(+)-carfentrazone, R-(-)-carfentrazone, S-(-)-carfentrazone-ethyl, and R-(+)-carfentrazone-ethyl, respectively. The specificity, matrix effect, linearity, precision, accuracy, and stability were surveyed to evaluate the feasibility of the method. The mean recovery was in the range of 77.5-102.8% with relative standard deviations of 0.4-9.8% for the samples. The limits of detection of the enantiomers were evaluated as 0.7 to 6.0 μg/kg, and the limits of quantification were 2.5 to 20 μg/kg. The stereoselective degradation of carfentrazone-ethyl and carfentrazone in rice plant was investigated, and there was no clear enantioselectivity for carfentrazone-ethyl. As for carfentrazone, the enantiomer fractions value reached 0.85 at 7 days after spraying. The developed method was simple and reliable enough for the corresponding risk assessment of carfentrazone-ethyl and its metabolite enantiomers in crop plants, cereal grains, and soil samples.

No abstract

R-(+)-carfentrazone-ethyl and S-(+)-carfentrazone was found to be degraded preferentially in the Jiangxi, Jilin and Anhui soils for the first time.

Enhanced lateral flow assay with double conjugates for the detection of exosomes

With the continuous development of computers, the gradual opening of the financial market, and China's increasing investment in scientific and technological innovation, artificial intelligence and big data analysis and other technologies have begun to be used in various financial industries such as the stock and bond market. This paper analyzes the correlation of stock indexes in Istanbul and other countries through python data analysis, and finds that the stock markets in various countries have relatively large correlation. The regression prediction analysis of Istanbul stock data is carried out, and three models of linear regression, ridge regression and gradient regression are constructed for prediction, and the performance comparison is made.