Exploration of GH94 Sequence Space for Enzyme Discovery Reveals a Novel Glucosylgalactose Phosphorylase Specificity

The substantial increase in DNA sequencing efforts has led to a rapid expansion of available sequences in glycoside hydrolase families. The ever-increasing sequence space presents considerable opportunities for the search for enzymes with novel functionalities. In this work, the sequence-function space of glycoside hydrolase family 94 (GH94) was explored in detail, using a combined approach of phylogenetic analysis and sequence similarity networks. The identification and experimental screening of unknown clusters led to the discovery of an enzyme from the soil bacterium Paenibacillus polymyxa that acts as a 4-O-β-d -glucosyl-d -galactose phosphorylase (GGalP), a specificity that has not been reported to date. Detailed characterization of GGalP revealed that its kinetic parameters were consistent with those of other known phosphorylases. Furthermore, the enzyme could be used for production of the rare disaccharides 4-O-β-d -glucosyl-d -galactose and 4-O-β-d -glucosyl-l -arabinose. Our current work highlights the power of rational sequence space exploration in the search for novel enzyme specificities, as well as the potential of phosphorylases for rare disaccharide synthesis.     

Portrait of Cancer Stem Cells on Colorectal Cancer: Molecular Biomarkers,Signaling Pathways and miRNAome

Colorectal cancer (CRC) is a leading cause of cancer death worldwide, and about 20% is metastatic at diagnosis and untreatable. Increasing evidence suggests that the heterogeneous nature of CRC is related to colorectal cancer stem cells (CCSCs), a small cells population with stemness behaviors and responsible for tumor progression, recurrence, and therapy resistance. Growing knowledge of stem cells (SCs) biology has rapidly improved uncovering the molecular mechanisms and possible crosstalk/feedback loops between signaling pathways that directly influence intestinal homeostasis and tumorigenesis. The generation of CCSCs is probably connected to genetic changes in members of signaling pathways, which control self-renewal and pluripotency in SCs and then establish function and phenotype of CCSCs. Particularly, various deregulated CCSC-related miRNAs have been reported to modulate stemness features, controlling CCSCs functions such as regulation of cell cycle genes expression, epithelial-mesenchymal transition, metastasization, and drug-resistance mechanisms. Primarily, CCSC-related miRNAs work by regulating mainly signal pathways known to be involved in CCSCs biology. This review intends to summarize the epigenetic findings linked to miRNAome in the maintenance and regulation of CCSCs, including their relationships with different signaling pathways, which should help to identify specific diagnostic, prognostic, and predictive biomarkers for CRC, but also develop innovative CCSCs-targeted therapies.     

3D CaP porous scaffolds with grooved surface topography obtained by the sol-gel method

The influence of surface topography on cellular behaviour and its importance for the development of three-dimensional scaffolds for bone tissue engineering are a topic of growing interest. To date, the introduction of topographical patterns into the surface of 3D porous ceramic scaffolds has proven difficult, due partly to the brittle nature of ceramic materials as well as the currently available fabrication technologies. In this study, a grooved pattern was introduced into the surface of 3D multilayer porous ceramic scaffolds by the chemical etching technique. The patterned scaffolds were characterised by X-Ray Diffraction (XRD), Scanning Electron Microscopy with Energy Dispersive X-Ray Spectroscopy (SEM-EDX) and Digital Holographic Microscopy (DHM). Their bioactivity was also evaluated in vitro by immersion in simulated body fluid (SBF) for 12 h, 1, 7, 14 and 21 days. Scaffolds were constituted mainly with a mixture of the calcium pyrophosphate (Ca₂O₇P₂) and β-tricalcium phosphate (Ca?(PO?)?) phases. The pyrophosphate on the external layer was dissolved as a result of the etching process, leaving grooves on the surface. Ridges and grooves were nano-/micrometric, with dimensions of around 900 nm–1.5 μm in width and 200 nm–300 nm in depth. Moreover, the mechanical properties and bioactive capacity of the patterned scaffolds were not affected by chemical etching, making them suitable to be used in bone tissue engineering.     

稳定同位素技术在禽类及其制品溯源领域的研究进展

本文基于国内外禽类及其制品溯源的研究成果，针对市场上出现的禽类及其制品的产地和饲养方式造假（如普通鸡蛋冒充地理标志鸡蛋、自由散养鸡蛋、有机鸡蛋等）、禽类饲料成分掺假（如非法添加动物副产品、色素等）等问题，梳理了稳定同位素技术在禽类及其制品可追溯性的应用进展，在此基础上提出禽类及其制品溯源的现存问题和发展方向，以期为稳定同位素技术在禽类及其制品质量安全领域的深入研究及食品溯源体系完善提供参考。     

Algorithm selection and instance space analysis for curriculum-based course timetabling

Journal of Scheduling - We propose an algorithm selection approach and an instance space analysis for the well-known curriculum-based course timetabling problem (CB-CTT), which is an important...     

南水北调中线总干渠藻类的生态调度

南水北调中线总干渠无在线调蓄水库,对藻类生态调度过程中出现的问题开展生态调度实现策略和实施方式研究。主要实现策略包括:划定自身的调蓄区,隔离生态调度对下游的影响;采用高效的渠池运行方式,减少生态调度时蓄量的反复调整;综合考虑安全、快速、平稳等需求,设定生态调度实施进程和方式。具体实施方式包括:将总干渠划分为流速调控区、调蓄区和正常运行区,分别实施等体积、控制蓄量和闸前常水位方式运行;将生态调度过程划分为充水阶段和泄水阶段,基于流速调控目标值、持续时长和水位降幅约束条件,确定各阶段时长和各分区的闸门群调控方案等。基于2018年3月输水工况,采用明渠一维非恒定流模型,仿真总干渠上游15个渠池的藻类生态调度过程。结果表明,生态调度可在3.5 d内完成,各渠池的平均流速由0.48 m/s增至0.93 m/s,持续时间超过2 h。在整个生态调度过程中,水位变化平稳,水位变幅符合安全阈值要求,下游渠道的正常运行未受生态调度明显影响。     

QoS metrics-in-the-loop for endowing runtime self-adaptation to robotic software architectures

Multimedia Tools and Applications - The design of robots capable of operating autonomously in changing and unstructured environments, requires using complex software architectures in which,...     

Implication of the NLRP3 Inflammasome in Bovine Age-Related Sarcopenia

Sarcopenia is defined as the age-related loss of skeletal muscle mass, quality, and strength. The pathophysiological mechanisms underlying sarcopenia are still not completely understood. The aim of this work was to evaluate, for the first time, the expression of NLRP3 inflammasome in bovine skeletal muscle in order to investigate the hypothesis that inflammasome activation may trigger and sustain a pro-inflammatory environment leading to sarcopenia. Samples of skeletal muscle were collected from 60 cattle belonging to three age-based groups. Morphologic, immunohistochemical and molecular analysis were performed to assess the presence of age-related pathologic changes and chronic inflammation, the expression of NLRP3 inflammasome and to determine the levels of interleukin-1β, interleukin-18 and tumor necrosis factor alpha in muscle tissue. Our results revealed the presence of morphologic sarcopenia hallmark, chronic lymphocytic inflammation and a type II fibers-selective NLRP3 expression associated to a significant decreased number of immunolabeled-fibers in aged animals. Moreover, we found a statistically significant age-related increase of pro-inflammatory cytokines such as interleukin-1β and interleukin-18 suggesting the activation of NLRP3 inflammasome. Taken together, our data suggest that NLRP3 inflammasome components may be normally expressed in skeletal muscle, but its priming and activation during aging may contribute to enhance a pro-inflammatory environment altering normal muscular anabolism and metabolism.