FRET-Based Mito-Specific Fluorescent Probe for Ratiometric Detection and Imaging of Endogenous Peroxynitrite: Dyad of Cy3 and Cy5

Peroxynitrite (OONO^–) is profoundly implicated in health and disease. The physiological and pathological outcome of OONO^– is related to its local concentration, and hence, a reliable OONO^– assay is highly desired. We have developed a FRET-based small-molecule fluorescent probe (<b>PNCy3Cy5</b>), harnessing the differential reactivity of Cy3 and Cy5 toward OONO^– by fine-tuning. It exhibits high detection sensitivity and yields a ratiometric fluorescent signal. We have exemplified that it can be applied in semiquantitative determination of OONO^– in living cells. Notably, it specifically localizes in mitochondria, where endogenous OONO^– is predominantly generated. Thus, <b>PNCy3Cy5</b> is a promising molecular tool for peroxynitrite biology

DataSheet_1_A population-based predictive model identifying optimal candidates for primary and metastasis resection in patients with colorectal cancer with liver metastatic.zip

BackgroundThe survival benefit of primary and metastatic resection for patients with colorectal cancer with liver metastasis (CRLM) has been observed, but methods for discriminating which individuals would benefit from surgery have been poorly defined. Herein, a predictive model was developed to stratify patients into sub-population based on their response to surgery.MethodsWe assessed the survival benefits for adults diagnosed with colorectal liver metastasis by comparing patients with curative surgery vs. those without surgery. CRLM patients enrolled in the Surveillance, Epidemiology, and End Results (SEER) database between 2004 and 2015 were identified for model construction. Other data including CRLM patients from our center were obtained for external validation. Calibration plots, the area under the curve (AUC), and decision curve analysis (DCA) were used to evaluate the performance of the nomogram compared with the tumor–node–metastasis (TNM) classification. The Kaplan–Meier analysis was performed to examine whether this model would distinguish patients who could benefit from surgery.ResultsA total of 1,220 eligible patients were identified, and 881 (72.2%) underwent colorectal and liver resection. Cancer-specific survival (CSS) for the surgery group was significantly better than that for the no-surgery group (41 vs. 14 months, p ConclusionsAn accurate and easy-to-use CRLM nomogram has been developed and can be applied to identify optimal candidates for the resection of primary and metastatic lesions among CRLM patients.</p

Monthly percentage of ILI outpatients positive for viral etiology in Shanghai, 2011–2013.

<p>A shows the monthly percentage of ILI outpatients positive for influenza virus, non-influenza virus and positive for any one of the studied respiratory viruses. B shows the monthly percentage of ILI outpatients positive for influenza subtypes. C shows the monthly percentage of ILI outpatients positive for HRV, HADV and HCoV. D shows the monthly percentage of ILI outpatients positive for HPIV, RSV and other respiratory viruses.</p

Additional file 1 of Prioritization of therapeutic targets for cancers using integrative multi-omics analysis

Additional file 1. Table S1. The information of outcomes resources. Table S2. The information of exposures resources. Table S3. The information of TWAS-significant genes in whole blood. Table S4. Results of SMR for genes associated with cancers in whole blood. Table S5. Results of colocalization for the genes signficantly associated wtith cancers in TWAS and SMR analysis in whole blood. Table S6. The information of TWAS-significant genes in specific organ tissues. Table S7. Results of SMR for genes associated with cancers in specific organ tissues. Table S8. Results of colocalization for the genes signficantly associated wtith cancers in TWAS and SMR analysis in specific organ tissues. Table S9. The information of PWAS-significant genes in whole blood. Table S10. Results of MR for protein-code genes associated with cancers in whole blood. Table S11. Results of colocalization for protein-code genes signficantly associated wtith cancers in PWAS and MR analysis in whole blood. Table S12. The list of druggable genes for SMR.  Table S13. Results of SMR for druggable genes associated with cancers. Table S14. Results of colocalization for identified druggable genes wtith cancers in FinnGen cohort. Table S15. Results of phenotype scanning on candidate genes. Table S16. The information of enrichment analysis for candidate genes. Table S17. Results of enrichment analysis for candidate genes. Table S18. The information of metabolic pathways. Table S19. The information of IVs for Metabolome-wide Mendelian Randomization on cancers. Table S20. Results of Metabolome-Wide Mendelian Randomization on cancers

Selective and Ratiometric Fluorescent Trapping and Quantification of Protein Vicinal Dithiols and in Situ Dynamic Tracing in Living Cells

Protein vicinal dithiols play fundamental roles in intracellular redox homeostasis due to their involvement in protein synthesis and function through the reversible vicinal dithiol oxidation to disulfide. To provide quantitative information about the global distribution and dynamic changes of protein vicinal dithiols in living cells, we have designed and synthesized a ratiometric fluorescent probe (<b>VTAF</b>) for trapping of vicinal dithiol-containing proteins (VDPs) in living cells. <b>VTAF</b> exhibits a ratiometric fluorescence signal upon single excitation, which enables self-calibration of the fluorescence signal and quantification of endogenous vicinal dithiols of VDPs. Its potential for in situ dynamic tracing of changes of protein vicinal dithiols under different cellular redox conditions was exemplified. <b>VTAF</b> facilitated the direct observation of subcellular distribution of endogenous VDPs via ratiometric fluorescence imaging and colocalization assay. And the results suggested that there are abundant VDPs in mitochondria. Moreover, some redox-sensitive VDPs are also present on cell surface which can respond to redox stimulus. This ratiometric fluorescence technique presents an important extension to previous fluorescence intensity-based probes for trapping and quantifying protein vicinal dithiols in living cells, as well as its visible dynamic tracing of VDPs

Baseline characteristics of patients with parathyroidectomy.

<p>Baseline characteristics of patients with parathyroidectomy.</p

Effects of PTH withdrawal on mineralization.

<p>Mineralized nodule formation was assessed by Alizarin red S (ARS) staining 20 days after differentiation. The blank control group without the supplement of bPTH and the osteogenic induction in the cell culture was performed as a negative control. Original magnification 100×.</p

Effects of PTH withdrawal on osteoblast proliferation.

<p>Cells were grown in microtiter plates in a final volume of 200 μL culture medium per well for 2–20 days. MTT assays were performed after 48 h. (A) Cell proliferation appears to be downregulated in the first 8 days, and inhibited after 10 days of culture in the continuously-treated bPTH groups. The effect of inhibition increases with the concentration of bPTH. (B) Although osteoblast proliferation coordinately declined in the PTH-C 100 ng/mL and PTH Day 1–10 groups, it rebounded after bPTH withdrawal in the PTH Day 1–10 group. *Significantly different compared to the PTH-C 100 ng/mL group at the same time point, <i>P</i> < 0.05.</p

Effects of PTH withdrawal on calcium and phosphorus in culture.

<p>Calcium and phosphorus derived from control and bPTH treated cultures at different time points. In the control group, there was a gradual decline of calcium and phosphorus content that reached a plateau after day 16. Similarly, calcium and phosphorus content slowly decreased with time in continuous bPTH cultures. Although the initial calcium and phosphorus content in cultures treated with transient bPTH decreased more slowly than in controls, they showed a rapid decline after bPTH withdrawal on day 10 and attained similar levels as control. *^{, #}Significantly different compared with control at the same time point, <i>P</i>< 0.05.</p

Effects of PTH withdrawal on AKP activity.

<p>In control cultures, AKP activity increased with time and reached a plateau after 14 days. In contrast, AKP activity declined from day 4 to day 10 and did not increase over time in the continuous PTH cultures. In the PTH Day 1–10 group that was treated with bPTH on days 1 to 10, AKP activity rebounded after withdrawal of bPTH and was significantly higher than controls and continuously bPTH-treated cells from day 16. AKP activity did not fully recover until day 20. *^{, #}Significantly different compared to control at the same time point, <i>P</i> < 0.05.</p