A feasible protocol to profile bile acids in dried blood spots from rats using a UHPLC-MS/MS method combining a surrogate matrix.

Dried blood spot (DBS) sampling is a promising method for microliter blood sample collection with the advantages of convenient transportation, storage and clinical operations. However, it is challenging to develop an analytical protocol to determine endogenous metabolites, such as bile acids (BAs) in DBSs, due to the low-blood-volume character of DBSs and the complex features of filter paper. Herein, we developed a method of fast ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) to profile and quantify BAs in DBSs. The pretreatment methods were optimized and a two-step solvent addition method, where a small amount of water was firstly added to moisten the DBS and then methanol was added, showed high extraction efficiency for multiple BAs in DBSs. The UHPLC-MS/MS conditions were optimized and 35BAs in different types could be profiled with good resolution and quantified with acceptable precision and accuracy. Preparation of a DBS surrogate matrix without endogenous BAs has been well developed using rat erythrocytes in BSA solution and showed good performance on both the signal suppression/enhancement percentage and parallelism assessment evaluation of three different stable-isotope-labeled (SIL) BAs. The established protocol was successfully applied to profile BAs in DBSs of intrahepatic cholestasis model and healthy control rats with good repeatability. To our knowledge, it is the first time that 35 BAs in DBSs could be well profiled and an appropriate DBS surrogate matrix has been developed. This protocol presents future-oriented applications of DBSs for relevant preclinical studies to profile BAs and probe biomarkers.

Evaluating the Effects of Built Environment on Street Vitality at the City Level: An Empirical Research Based on Spatial Panel Durbin Model.

There is evidence that the built environment has an influence on street vitality. However, previous studies seldom assess the direct, indirect, and total effect of multiple environmental elements at the city level. In this study, the features of the street vitality on Xiamen Island are described based on the location-based service Big Data. Xiamen Island is the central urban area of Xiamen, one of the national central cities in China. With the help of multi-source data such as street view images, the condition of design that is difficult to effectively measure with traditional data can be better explored in detail on a macro scale. The built environment is measured through a 5D system at the city level, including Density, Diversity, Design, Destination accessibility, and Distance to transit. Spatial panel Durbin models are constructed to analyze the influence of the built environment on the street vitality on weekdays and weekends, and the direct, indirect, and total effects are evaluated. Results indicate that at the city level, the built environment plays a significant role in promoting street vitality. Functional density is not statistically significant. Most of the elements have spatial effects, except for several indicators in the condition of the design. Compared with the conclusions of previous studies, some indicators have different effects on different spatial scales. For instance, on the micro scale, greening can enhance the attractiveness of streets. However, on the macro scale, too much greening brings fewer functions along the street, which inhibits the street vitality. The condition of design has the greatest effect, followed by destination accessibility. The differences in the influences of weekdays and weekends are mainly caused by commuting behaviors. Most of the built environment elements have stronger effects on weekends, indicating that people interact with the environment more easily during this period.

Genomics-driven discovery of a new cyclodepsipeptide from the guanophilic fungus Amphichorda guana.

Two potential non-ribosomal peptide synthetases (NRPSs) were identified in the genome of a guanophilic fungus Amphichorda guana by bioinformatics analysis and gene knockout experiments. Liquid chromatography coupled with mass spectrometry (LC-MS) guided isolation led to the discovery of a new cyclodepsipeptide isaridin H (1) and seven known analogs, desmethylisaridin E (2), isaridin E (3), isariin A (4), iso-isariin B (5), iso-isariin D (6), isariin E (7), and nodupetide (8). The absolute configuration of isaridin H (1) was achieved by Marfey's method. Isaridin H (1) showed significant antifungal activity against Botrytis cinerea and Alternaria solani.

Comparative Genomic Data Provide New Insight on the Evolution of Pathogenicity in Sporothrix Species.

Sporothrix species are commonly isolated from environmental and clinical samples. As common causes of zoonotic mycosis, Sporothrix species may result in localized or disseminated infections, posing considerable threat to animal and human health. However, the pathogenic profiles of different Sporothrix species varied, in virulence, geographic location and host ranges, which have yet to be explored. Analysing the genomes of Sporothrix species are useful for understanding their pathogenicity. In this study, we analyzed the whole genome of 12 Sporothrix species and six S. globosa isolates from different clinical samples in China. By combining comparative analyses with Kyoto Encyclopedia of Genes and Genomes (KEGG), Carbohydrate-Active enZymes (CAZy), antiSMASH, Pfam, and PHI annotations, Sporothrix species showed exuberant primary and secondary metabolism processes. The genome sizes of four main clinical species, i.e., S. brasiliensis, S. schenckii, S. globosa, and S. luriei were significantly smaller than other environmental and clinical Sporothrix species. The contracted genes included mostly CAZymes and peptidases genes that were usually associated with the decay of plants, as well as the genes that were associated with the loss of pathogenicity and the reduced virulence. Our results could, to some extent, explain a habitat shift of Sporothrix species from a saprobic life in plant materials to a pathogenic life in mammals and the increased pathogenicity during the evolution. Gene clusters of melanin and clavaric acid were identified in this study, which improved our understanding on their pathogenicity and possible antitumor effects. Moreover, our analyses revealed no significant genomic variations among different clinical isolates of S. globosa from different regions in China.