Preparation and Characterization of an Ancient Aminopeptidase Obtained from Ancestral Sequence Reconstruction for L-Carnosine Synthesis

As a biologically active peptide, L-carnosine has been widely used in the pharmaceutical, cosmetic and health care industries due to its various physiological properties. However, relatively little research is available regarding L-carnosine’s enzymatic synthesis function. In this study, a potential enzyme sequence with the function of carnosine synthesizing was screened out using the ancestral sequence reconstruction (ASR) technique. Identified with L-carnosine synthesis activity, this enzyme was further confirmed using autoproteolytic phenomenon via Western blot and N-terminal sequencing. After purification, the enzymatic properties of LUCA–DmpA were characterized. The melting temperature (T_m) and denaturation enthalpy (ΔH) of LUCA–DmpA were 60.27 ± 1.24 °C and 1306.00 ± 26.73 kJ·mol⁻¹, respectively. Circular dichroism (CD) spectroscopy results showed that this ancestral enzyme was composed of α-helix (35.23 ± 0.06%), β-sheet (11.06 ± 0.06%), β-turn (23.67 ± 0.06%) and random coil (32.03 ± 0.06%). The enzyme was characterized with the optimal temperature and pH of 45 °C and 9.0, respectively. Notably, LUCA–DmpA was also characterized with remarkable pH tolerance based on the observation of more than 85% remaining enzymatic activity after incubation at different pH buffers (pH = 6–11) for 12 h. Additionally, rather than being improved or inhibited by metal ions, its enzymatic activity was found to be promoted by introducing organic solvent with a larger log P value. Based on these homology modeling results, the screened LUCA–DmpA is suggested to have further optimization potential, and thereafter to be offered as a promising candidate for real industrial applications.     

Atomic-scale simulations for lithium dendrite growth driven by strain gradient

Dendrite formation is a major obstacle, e.g., capacity loss and short circuit,to the next-generation high-energy-density lithium(Li)-metal batteries. The development of successful Li dendrite mitigation strategies is impeded by an insu?cient understanding in Li dendrite growth mechanisms. The Li-plating-induced internal stress in Li-metal and its e?ects on dendrite growth have been widely studied, but the underlying microcosmic mechanism is elusive. In the present study, the role of the plating-...     

离散梯度法在基于图像的计算生物力学中的应用1)

介绍一种可用于计算生物力学的离散梯度方法,此方法可利用离散的点云模型直接进行数值模拟分析而不需要传统的几何模型。将离散梯度法应用于点云模型需要首先确定模型中点之间的相邻关系和每个点所分配的材料体积,然后通过用广义的有限差分的形式定义了梯度插值向量,并以此向量来近似函数在每个离散点上的梯度。从弱形式出发,推导建立了适用于弹性固体大变形问题的求解器,并具有和有限元法中双线性四边形单元一致的准确性和收敛性。着重描述了一种可以从医学图像中快速提取材料点并建立点云模型的方法,以及利用三角划分和重心划分确定材料点之间的相邻关系和每个材料点体积的具体过程。通过腹主动脉瘤膨胀的静力学模拟分析,展示了离散梯度法的实用性和准确性。该算法实现了基于医学图像进行生物力学分析的过程自动化,为病体特异性的研究和治疗提供便利和实用的工具。     

缺口件多轴疲劳寿命预估新方法

局部应力应变法(LSSM)以危险点处的局部应力应变历程进行寿命评价,忽略了非比例附加强化及缺口附近应力梯度对疲劳损伤的影响.论文通过分析裂纹萌生面(临界面)上剪应力和正应力分布状态,提出一种计算缺口件多轴疲劳损伤影响区的数学计算方法,建立考虑法向正/剪应力梯度与非比例附加强化效应的寿命预估模型.首先,基于能量法和临界面理论,借助坐标变换原理将应变能密度最大的材料平面定义为临界面,建立确定裂纹萌生方向的计算方法.其次,以临界面上特定路径为积分方向,将缺口件三维问题转化为线性问题,简化计算过程.综合考虑峰值应力和等效应力场强对多轴疲劳寿命的贡献度,建立表征缺口件多轴疲劳损伤演化过程的损伤参量.最后,通过TC4合金和Q345钢多轴疲劳试验验证论文模型的可行性和精确度,并与LSSM法、FS模型、SWT模型进行对比分析.     

高功率光学元件校正波前的梯度均方根指标

对高功率固体激光装置中,光学元件波前经过变形镜校正后的梯度均方根指标开展了研究。建立了变形镜校正效果的等效分解方法,分析了校正波前的部分相干叠加特性,对校正波前的梯度均方根与焦斑尺寸的对应关系进行了研究。解决了在有变形镜校正时,如何制定元件波前梯度均方根指标的问题。研究结果对高功率固体激光装置的光学元件波前指标制定有指导意义。     

光梯度力与激光捕获

从电偶极子在非均匀电磁场中所受梯度力出发,用光的电磁理论和电解质小球的电偶极子模型,论述了介质小球在光场中所受光梯度力与光强梯度成正比,并以此来定量说明激光对介质小球的捕获原理.     

阵列微流控浓度梯度网络用于细胞-化学刺激反应研究

设计和制作了具有5组平行浓度梯度形成网络和30个细胞培养池的微流控芯片,该芯片集成了细胞接种、培养、梯度浓度化学刺激、标记和检测等功能单元。芯片为玻璃-PDMS杂合结构,微流控通道刻蚀于玻璃层。芯片细胞培养池设计了系列围堰结构以利于细胞贴壁。细胞接种、灌流培养和试剂引入通过外接微量注射泵控制完成。该芯片可以生成连续、稳定的平行浓度梯度。观察发现,围堰结构有利于细胞接种和生长,乳腺癌MCF-7细胞在芯片灌流培养条件下生长良好。利用该芯片检测了在接受As2O3和乙酰丝氨酸(NAC)梯度浓度刺激后乳腺癌MCF-7细胞内谷胱甘肽(GSH)水平以及细胞阿霉素敏感性的变化,分析乳腺癌细胞阿霉素敏感性与细胞内GSH水平的关系。MCF-7细胞内GSH水平的变化与刺激药物浓度呈剂量-效应依赖关系,在接受As2O3刺激后GSH水平有所下降;而在接受NAC刺激后GSH水平有所升高。MCF-7细胞阿霉素敏感性与GSH水平相关。在降低GSH水平后药物敏感性升高;而升高细胞内GSH水平后敏感性降低。这种阵列微流控浓度梯度网络可以用于高通量细胞-化学刺激反应研究,有潜力成为细胞水平大规模药物筛选的技术平台。     

基于改进NAS-RIF的三维显微图像盲复原

对二维NAS-RIF算法进行了改进,并将它用于三维显微图像复原,分别用模拟样本和真实生物样本进行了实验。针对原算法对噪声有放大作用,提出了基于中值滤波去噪的改进。在支持域的确定、最小点的搜索上进行改进,提高复原质量。改进了原算法仅适用于均匀背景的不足,扩大了应用范围。实验结果表明,该算法在三维显微图像盲复原中具有较好的效果。     

基于解码技术的全息显微术研究

提出了一种利用解码技术对全息显微畸变图像进行校正的方法。把全息记录中的显微镜预放大过程作为物光波产生畸变的编码过程;在波前再现过程中,用全息记录中的预放大显微镜对再现的赝实像再次放大进行解码,从而获得清晰的全息显微图像。实验验证了该方法的有效性。     

Spatio-Temporal Variation of the Bacterial Communities along a Salinity Gradient within a Thalassohaline Environment (Saline di Tarquinia Salterns,Italy)

The “Saline di Tarquinia” salterns have been scarcely investigated regarding their microbiological aspects. This work studied the structure and composition of their bacterial communities along the salinity gradient (from the nearby sea through different ponds). The communities showed increasing simplification of pond bacterial diversity along the gradient (particularly if compared to those of the sea). Among the 38 assigned phyla, the most represented were Proteobacteria, Actinobacteria and Bacteroidetes. Differently to other marine salterns, where at the highest salinities Bacteroidetes dominated, preponderance of Proteobacteria was observed. At the genus level the most abundant taxa were Pontimonas, Marivita, Spiribacter, Bordetella, GpVII and Lentibacter. The α-diversity analysis showed that the communities were highly uneven, and the Canonical Correspondence Analysis indicated that they were structured by various factors (sampling site, sampling year, salinity, and sampling month). Moreover, the taxa abundance variation in relation to these significant parameters were investigated by Generalized Linear Models. This work represents the first investigation of a marine saltern, carried out by a metabarcoding approach, which permitted a broad vision of the bacterial diversity, covering both a wide temporal span (two years with monthly sampling) and the entire salinity gradient (from the nearby sea up to the crystallisation ponds).