Heterologous protein production capacity of mammalian cells cultivated as monolayers and microtissues

A precise understanding of processes managing heterologous protein production in vitro and in vivo is essential for the manufacture of sophisticated biopharmaceuticals as well as for future gene therapy and tissue engineering initiatives. Capitalizing on the gravity-enforced self-assembly of monodispersed cells into coherent (multicellular) microtissues we studied heterologous protein production of microtissues and monolayers derived from cell lines and primary cells engineered/transduced for (i) constitutive, (ii) proliferation-controlled, (iii) macrolide-, or (iv) gas-inducible expression of the human placental secreted alkaline phosphatase (SEAP) and of the Bacillus stearothermophilus-derived secreted alpha-amylase (SAMY). Specific productivity of cells assembled in microtissues was up to 20-fold higher than isogenic monolayer cultures. Diffusion across microtissues could be further increased by HUVEC-mediated vascularization. As well as higher specific protein productivities, microtissues were also more efficient than monolayer cultures in assembling transgenic lentiviral particles. Our results showed that mammalian cells embedded in a tissue-like three-dimensional (3D) microenvironment exhibit increased production capacity. This observation should be considered for gene therapy and tissue engineering scenarios as well as for biopharmaceutical manufacturing.     

Genomic analysis of a novel integrative conjugative element in Vibrio cholerae

Integrative conjugative elements (ICEs) are a class of self-transmissible mobile elements that mediate horizontal gene transfer in bacteria, and play an important role in bacterial evolution. Since 1992, ICEs of the SXT/R391 family have been found to be widely distributed among Vibrio cholerae strains isolated in Asian countries. Here we describe ICEVchB33, an ICE found in the genomes of two V. cholerae O1 Eltor strains, one isolated in India, 1994, and the other from Mozambique, 2004. ICEVchB33 revealed a new genetic organization, different from other ICEs of the SXT/R391 family, demonstrating the genomic plasticity of these elements.     

Engineering microenvironments for manufacturing therapeutic cells

There are a growing number of globally approved products and clinical trials utilizing autologous and allogeneic therapeutic cells for applications in regenerative medicine and immunotherapies. However, there is a need to develop rapid and cost-effective methods for manufacturing therapeutically effective cells. Furthermore, the resulting manufactured cells may exhibit heterogeneities that result in mixed therapeutic outcomes. Engineering approaches that can provide distinct microenvironmental cues to these cells may be able to enhance the growth and characterization of these cell products. This mini-review describes strategies to potentially enhance the expansion of therapeutic cells with biomaterials and bioreactors, as well as to characterize the cell products with microphysiological systems. These systems can provide distinct cues to maintain the quality attributes of the cells and evaluate their function in physiologically relevant conditions.     

The therapeutic potential of stem cells

In recent years, there has been an explosion of interest in stem cells, not just within the scientific and medical communities but also among politicians, religious groups and ethicists. Here, we summarize the different types of stem cells that have been described: their origins in embryonic and adult tissues and their differentiation potential in vivo and in culture. We review some current clinical applications of stem cells, highlighting the problems encountered when going from proof-of-principle in the laboratory to widespread clinical practice. While some of the key genetic and epigenetic factors that determine stem cell properties have been identified, there is still much to be learned about how these factors interact. There is a growing realization of the importance of environmental factors in regulating stem cell behaviour and this is being explored by imaging stem cells in vivo and recreating artificial niches in vitro. New therapies, based on stem cell transplantation or endogenous stem cells, are emerging areas, as is drug discovery based on patient-specific pluripotent cells and cancer stem cells. What makes stem cell research so exciting is its tremendous potential to benefit human health and the opportunities for interdisciplinary research that it presents.     

植物区系基本特征的参数综合表达

确定了植物区系６个基本特征的数量描述与处理方法,应用模糊数学中综合评判的方法建立了植物区系基本特征的参数综合表达式,最后通过实例验证,其参数综合表达式计算出来的实际情况一致。     

Hub genes identification and association of key pathways with hypoxia in cancer cells: A bioinformatics analysis

Three human cancer cell lines (A549, HCT116, and HeLa) were used to investigate the molecular mechanisms and potential prognostic biomarkers associated with hypoxia. We obtained gene expression data from Gene Expression Omnibus (GEO) datasets GSE11704, GSE147384, and GSE38061, which included 5 hypoxic and 8 control samples. Using the GEO2R tool and Venn diagram software, we identified common differentially expressed genes (cDEGs). The cDEGs were then subjected to Gene ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis by employing DAVID. The hub genes were identified from critical PPI subnetworks through CytoHuba plugin and these genes' prognostic significance and expression were verified using Kaplan-Meier analysis and Gene Expression Profiling Interactive Analysis (GEPIA), respectively. The research showed 676 common DEGs (cDEGs), with 207 upregulated and 469 downregulated genes. The STRING analysis showed 673 nodes and 1446 edges in the PPI network. We identified 4 significant modules and 19 downregulated hub genes. GO analysis revealed all of them were majorly involved in ribosomal large subunit assembly and biogenesis, rRNA processing, ribosome biogenesis, translation, RNA & protein binding frequently at the sites of nucleolus and nucleoplasm while 11 were significantly associated with a better prognosis of hypoxic tumors. Our research sheds light on the molecular mechanisms that underpin hypoxia in human cancer cell lines and identifies potential prognostic biomarkers for hypoxic tumors.     

考洲洋饵料生物水平综合评价

在GIS(Geographical Information System)的支持下,利用等级模型对1999年2月(枯水期)和8月(丰水期)考洲洋饵料生物水平进行了综合的分析和评价。枯水期考洲洋整个水域饵料生物的分布较均匀,其密集分布区位于湾西北部、湾口和吉隆河口附近水域,达4级水平,饵料生物较为丰富。丰水期饵料生物水平的变化幅度较大,其密集分布区位于湾中部和望京洲沿岸水域,达4-5级,饵料生物最为丰富,其次为湾口,达4级水平。两个调查航次相比,丰水期的饵料生物水平高于枯水期。主要优势种的变化具有明显的季节交叉和演替现象,浮游植物和浮游动物的主要优势种在不同季节差异较大,而底栖生物的优势种组成在整个调查期间变化不大。调查期间各饵料生物的多样性指数和均匀度均呈现出较大的变化,其最大变化范围分别为0.01-4.49和0.01-0.96,表明湾内饵料生物受外界环境条件影响较大,生物群落组成结构和生物多样性水平不太稳定。调查期间整个水域的生物多样性阈值均呈现出湾西部及西北部水域较高,其余水域较低的分布特点,但不同季节其变化趋势及分布范围略有差异。枯水期多样性阈值的密集分布区出现于湾西侧的部分水域,其等级水平为3级,饵料生物多样性较好,其余水域均为2级或低于2级水平,多样性一般或较差。丰水期的分布趋势与枯水期相似,其密集区出现于湾西部和西北部的整片水域中,分布范围比枯水期大,但等级水平明显低于枯水期,为2级,多样性一般,其余大部分水域均为1级水平,多样性较差。       

CLIP-170S is a microtubule +TIP variant that confers resistance to taxanes by impairing drug-target engagement

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短期整体身心调节法的全身皮肤电导反应规律

目的：探讨短期整体身心调节法练习者的全身皮肤电导反应特点。方法：采用单因素完全随机实验设计,将68名在校大学生分为整体身心调节组和放松组,分别进行为期10天的整体身心调节训练和经典的肌肉放松训练,并于训练前和训练结束后,测试两组学生身体7个分支的皮肤电导反应。结果：训练前,大学生前3支皮肤电导反应大于后4支;训练10天后,与训练前相比,整体身心调节动I练组第1、2支皮肤电导反应显著减小,7个分支的皮肤电导反应出现均衡的规律;放松训练组学生身体7个分支皮肤电导反应变化不显著。结论：短期整体身心调节法形成了全身皮肤电导反应的均衡,可能反映短期整体身心调节法的整体身心相互作用原理。