Solvent-Driven Room-Temperature Curtius Rearrangements to Access Nucleotides Bearing Substituted Fused Pyridones.

The intramolecular Curtius rearrangement suffers from a high reaction temperature, low yields, tedious product isolation, and difficult scale up. This study presents a room-temperature Curtius rearrangement that can be novelly driven by the HFIP solvent, followed by light-illuminated intramolecular cyclization. Such a mild reaction allows for the preparation of various fused pyridone derivatives with diverse substituent groups that have rarely been incorporated by previous methods. The roles of HFIP and light are investigated by a set of control experiments through a combination of IR and NMR titration. Furthermore, using the substituted fused pyridones as unnatural bases, we can obtain a panel of new nucleotides.

The gut-lung axis: Gut microbiota changes associated with pulmonary fibrosis in mouse models induced by bleomycin.

The main objective of this study was to investigate the alterations in the gut microbiota (GM) of pulmonary fibrosis (PF) mice induced by bleomycin (BLM) with its underlying mechanisms. BLM was docked with the targets of TGF-ß/SMAD and caspase-3 pathways using the molecular docking technique. HE staining and Masson staining were applied to observe the histopathological changes in the pulmonary tissues. Detection of the apoptotic signals was conducted by flow cytometry and TUNEL staining. The mRNA expression of targets involved in the TGF-ß/SMAD and caspase-3 signaling pathways in lungs was determined by qPCR. Immunohistochemistry (IHC) assay was used to detect the expression levels of cleaved caspase-3 and BAX proteins in mice lung tissues. 16S rDNA sequencing analysis was used to investigate the changes of GM in the fecal samples of mice in each group. The results showed that the apoptosis rate of pulmonary cells in the BLM group distinctly increased, with the expression levels of crucial target pro-apoptotic gene caspase-3, BAX with the corresponding protein, cleaved caspase-3, BAX were apparently elevated. This was accompanied by a significant increase in pro-fibrotic targets level such as TGF-ß, fibronectin, collagen I, and collagen III. The mechanisms of PF induced by BLM were related to apoptosis of lung tissue cells such as alveolar epithelial cells and destroyed alveolar structure and excessive production of extracellular matrix (ECM), which may be bound up with activating TGF-ß/SMAD and caspase-3 pathways. As for the GM, it was found that, after BLM induced PF in mice, the micro ecological balance of the GM was destroyed; the distance of PCo1 and Pco2 was significantly elongated, and the relative abundance of some intestinal probiotics like Catenibacterium and Lactobacillus (L. johnsonii and L. gasseri) dramatically lowered while the relative abundance of Verrucomicrobiales and Enterobacteriales substantially increased. Therefore, GM changes associated with PF in mouse models induced by BLM and the concept of "gut-lung axis" might provide an optional therapeutic strategy for PF.

Effects of land use patterns on stream water quality: a case study of a small-scale watershed in the Three Gorges Reservoir Area, China.

In this study, we have considered the relationship between the spatial configuration of land use and water quality in the Three Gorges Reservoir Area. Using land use types, landscape metrics, and long-term water quality data, as well as statistical and spatial analysis, we determined that most water quality parameters were negatively correlated with non-wood forest and urban areas but were strongly positively correlated with the proportion of forest area. Landscape indices such as patch density, contagion, and the Shannon diversity index were able to predict some water quality indicators, but the mean shape index was not significantly related to the proportions of farmland and water in the study area. Regression relationships were stronger in spring and fall than in summer, and relationships with nitrogen were stronger than those of the other water quality parameters (R(2) > 0.80) in all three seasons. Redundancy analysis showed that declining stream water quality was closely associated with configurations of urban, agricultural, and forest areas and with landscape fragmentation (PD) caused by urbanization and agricultural activities. Thus, a rational land use plan of adjusting the land use type, controlling landscape fragmentation, and increasing the proportion of forest area would help to achieve a healthier river ecosystem in the Three Gorges Reservoir Area (TGRA).

[Land Use Structure Change and Its Control Effect of Nitrogen Output in a Small Watershed of Three Gorges Reservoir Area: A Case Study of Lanlingxi Watershed].

The nitrogen (N) output in Lanlingxi watershed of Three Gorges Reservoir Area in 2015 was monitored and the current land use map was investigated. Cluster analysis and correlation analysis were used to identify the major sources of pollutants and to discriminate the source and the sink land use types. The effects of land use on N exports were quantitatively analyzed by stepwise regression analysis. The results showed that: ① After the returning farmland to forest project the land use structure of this area changed dramatically. The area proportions of forest land and garden plot increased to 76.85% and 13.87% respectively, and the proportion of cultivated land dropped to 1.16%. Cultivated lands were stellate distributed and garden plots in some catchments were flake distributed. ② The content of TN in some monitoring points surpassed the Class V standard of the national groundwater environmental quality. The ammonium-N (NH4+-N) concentrations ranged 0.089-0.214 mg·L-1, 2.925-13.203 mg·L-1 for nitrate-N (NO3--N) and 3.561-14.572 mg·L-1 for total-N (TN). And NO3--N accounted for more than 80% of TN. ③ There were significant positive correlations between garden plot, residential land and N loss, and negative correlations between forest land, unutilized land and N export, which indicated that the former were N sources and the latter were sinks. ④ Forest land area should be increased and garden plot should be controlled, and the area ratio of residential land should be kept below 5% when adjusting land use structure of this watershed. Moreover, forest-tea and forest-fruits modes should be applied to change single land use type of garden plot in some catchments.

[Effects of land use and landscape pattern on nitrogen and phosphorus exports in Lanlingxi Watershed of the Three Gorges Reservoir Area, China].

The temporal and spatial characteristics of N, P exports and effects of land use and landscape pattern on N, P exports were analyzed in the Lanlingxi Watershed of the Three Gorges Reservoir Area. The results showed that the TN, TP and NO3(-) -N were mainly generated by non-wood forest, the N, P exports in flood period (June to September) were significantly higher than the non-flood period (January to May). The NH4(+) -N export was derived from the residential area in the non-flood period, while from non-wood forest in the flood period. In addition, the performance of samples N, P exports with forest distributed were lower in both two periods. Also, the proportion of forest significantly negatively correlated with NO3(-) -N, TP in the non-flood period and TN, TP in the flood period. The residential area proportion notably positively correlated with NO3(-) -N, TN in non-flood period and NO3(-) -N, TN, TP in the flood period. The non-wood forest proportion also significantly positively correlated with NH4(+) -N, TN in the flood period. Moreover, PD closely positively correlated with N exports in non-flood period, with NO3(-) -N, NH4(+) -N in flood period. The CONT index strongly negatively correlated with N exports in flood period and TP in non-flood period. However, the proportions of farmland, unused land and the indices of ED were relatively weakened with N, P exports in both periods, while SHMN and water proportion did not show any positive or negative correlation. Moreover, the regression fitting degree of NH4(+)-N was superior to NO3(-) -N, TN and TP with the adjust R2 of 0.885 and 0.969 in two periods, while the regression relation was better than that of non-flood period. The result of redundancy analysis further demonstrated that the landscape fragmentation caused by patches types of different land uses could better explain impacts on the exports of nitrogen and phosphorus. The two canonical axes accumulated explained the 90% proportion of the variables and the highest contribution was PD, which was an important indicator for watershed water quality assessment and prediction.

RFP tags for labeling secretory pathway proteins.

Red fluorescent proteins (RFPs) are useful tools for live cell and multi-color imaging in biological studies. However, when labeling proteins in secretory pathway, many RFPs are prone to form artificial puncta, which may severely impede their further uses. Here we report a fast and easy method to evaluate RFPs fusion properties by attaching RFPs to an environment sensitive membrane protein Orai1. In addition, we revealed that intracellular artificial puncta are actually colocalized with lysosome, thus besides monomeric properties, pKa value of RFPs is also a key factor for forming intracellular artificial puncta. In summary, our current study provides a useful guide for choosing appropriate RFP for labeling secretory membrane proteins. Among RFPs tested, mOrange2 is highly recommended based on excellent monomeric property, appropriate pKa and high brightness.

A paper-based surface-enhanced resonance Raman spectroscopic (SERRS) immunoassay using magnetic separation and enzyme-catalyzed reaction.

In this study, a novel paper-based SERRS immunoassay based on magnetic separation and alkaline phosphatase (ALP) enzyme catalyzed hydrolysis reaction was developed. By using modified antibodies conjugated to magnetic beads, capture of mouse IgG followed by addition of ALP-labeled antibodies would form a sandwich-like immunoconjugate. After magnetic separation, 5-bromo-4-chloro-3-indolyl phosphate (BCIP), a low SERRS active compound, was added as the substrate for ALP to generate a high SERRS response. Detection was conducted on a silver colloid/PVP/filter paper SERS substrate by spotting a pre-aggregated silver colloid sol onto polyvinyl pyrrolidone (PVP) modified filter paper using a semi-automatic TLC sample applicator. The optimization of the highly SERS active paper-based substrate, dynamic hydrolysis process of BCIP, quantitative detection of IgG, and selectivity of the assay was studied in detail. By taking advantage of magnetic separation in order to decrease the background interference, the selective enzyme reaction involved in producing a highly SERRS active product could detect mouse IgG from 1 to 500 ng mL(-1) with a LOD of 0.33 ng mL(-1).