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For high-throughput screening (HTS) of Bacillus fastidiosus uricase mutants, a practical system was proposed. By error-prone PCR with final 1.5 mM MnCl, two focused libraries of mutants for A1-V158 and V150-D212 were generated separately. After induced expression of individual clones in 48-well microplates, Escherichia coli cells (BL21) were lyzed by 1.0 M Tris-HCl at pH 9.0 in 96-well microplates at 25 °C for 7.5 ~ 10.5 h; uricase reaction was continuously monitored with 0.15 mM uric acid in 96-well plates by absorbance at 298 nm to estimate V / K by kinetic analysis of reaction curve for comparison. V / K was resistant to initial uric acid levels with an upper limit 3-fold over that of initial rates. By receiver-operator-characteristic analysis of the recognition of the one of higher activity in uricase pair whose specific activity ratio was 1.8 or 3.3, the area-under-the-curve was comparable to that with cell lysates prepared by sonication treatment. A cutoff for the maximum Youden index was thus developed to recognize positive mutants of 1-fold higher activity. Indeed, mutant L171I/Y182F/Y187F/A193S of higher activity but lower thermostability at pH 7.4 and mutant V144A of higher activity and consistent thermostability were discovered. Therefore, the proposed system was practical for HTS of uricase mutants. [ABSTRACT FROM AUTHOR]

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A neural network Hamilton–Jacobi–Bellman (HJB) approach is introduced to deal with the spacecraft rendezvous problem with target spacecraft in arbitrary elliptical orbit. The Lawden equations are utilized to describe the relative motion of two spacecrafts. A generalized non-quadratic functional is introduced to describe constrained control. An approximate solution to the value function of the HJB equation corresponding to constrained controls is obtained by solving for a sequence of cost functions satisfying a sequence of Lyapunov equations. An inverse optimal controller is introduced to design the initial stabilizing admissible control for successive approximation. Furthermore, an optimal control law is obtained to stabilize the closed-loop system under constrained controls, and the spacecraft rendezvous mission can be accomplished with the nearly optimal controller. In comparison with the existing quadratic-regulation-based approaches used to deal with the rendezvous problem, which requires the value function of the nonlinear differential equations, the optimization factor and constrained control are taken into consideration simultaneously, and an approximate optimal constrained state feedback controller has been tuned a priori off-line. Stability analysis as well as simulation results are provided to illustrate the effectiveness of the presented approach. [ABSTRACT FROM AUTHOR]

Since in practice the line-of-sight (LOS) rate is difficult for a pursuer to measure accurately, the robust nonlinear H observer is applied to estimate it using LOS azimuths without any priori information of target maneuvers or its estimate in the presence of measurement noise. A threedimensional guidance law is implemented by replacing the LOS rate with their estimates using only relative range, speed, and LOS azimuths. The stability analyses and performed simulations results show that the presented approach is effective. [ABSTRACT FROM AUTHOR]

A series of novel 3-substituted amino-4-hydroxycoumarin derivatives have been designed and synthesized as chitin synthase (CHS) inhibitors. All the synthesized compounds have been screened for their CHS inhibition activity and antimicrobial activityin vitro. The enzymatic assay indicated that most of the compounds have good inhibitory activity against CHS, in which compound6owith IC50of 0.10 mmol/L had stronger activity than that of polyoxins B, which acts as control drug with IC50of 0.18 mmol/L. As far as the antifungal activity is concerned, most of the compounds possessed moderate to excellent activity against some representative pathogenic fungi. Especially, compound6bwas found to be the most potent agent againstCryptococcus neoformanswith minimal inhibitory concentration (MIC) of 4 μg/mL. Moreover, the results of antibacterial screening showed that these compounds have negligible actions to some tested bacteria. Therefore, these compounds would be promising to develop selective antifungal agents. [ABSTRACT FROM AUTHOR]

Esophageal cancer is of high prevalence and poor prognosis. Hesperetin has been reported to exert antitumor ability by inducing apoptosis in many cancers in vitro and in vivo without obvious toxicity. However, there is no study concerning about the effect of hesperetin on esophageal cancer. In this study, we aimed to investigate whether hesperetin could induce apoptosis in esophageal cancer cells and explore its potential mechanism. We found that hesperetin induced esophageal cancer cells apoptosis in a concentration-dependent and time-dependent manner compared with the untreated cells. Hoechst 33258 staining and flow cytometry analysis showed more apoptotic cells in the hesperetin-treated group ( p < 0.05, respectively). The intracellular reactive oxygen species (ROS) increased significantly, and glutathione (GSH) was depleted. The loss of △Ψ was more tremendous in the hesperetin-treated cells. N-acetylcysteine (NAC) reduced the proapoptotic ability of hesperetin, while DL-buthionine-S, R-sulfoximine (BSO) enhanced the anticancer effect. Western blotting showed that the expression levels of cytochrome C (Cyt C) and apoptosis-inducing factor (AIF) decreased in mitochondria and increased in cytoplasm ( p < 0.05). The levels of intracellular cleaved caspase-9, cleaved caspase-3, Apaf-1, Bcl-2-associated X protein (Bax), and suppressor of fused (SuFu) increased, while B cell lymphoma 2 (Bcl-2) and Survivin decreased. What is more, in xenograft tumor model, hesperetin inhibited the tumor growth significantly via induction of cell apoptosis which was detected by TUNEL assay ( p < 0.05). Taken together, our study demonstrated that hesperetin could induce cell apoptosis in esophageal cancer cells via mitochondrial-mediated intrinsic pathway by accumulation of ROS. [ABSTRACT FROM AUTHOR]