A rapid enzymatic assay for high-throughput screening of adenosine-producing strains

Adenosine is a major local regulator of tissue function and industrially useful as precursor for the production of medicinal nucleoside substances. High-throughput screening of adenosine overproducers is important for industrial microorganism breeding. An enzymatic assay of adenosine was developed by combined adenosine deaminase (ADA) with indophenol method. The ADA catalyzes the cleavage of adenosine to inosine and NH₃, the latter can be accurately determined by indophenol method. The assay system was optimized to deliver a good performance and could tolerate the addition of inorganic salts and many nutrition components to the assay mixtures. Adenosine could be accurately determined by this assay using 96-well microplates. Spike and recovery tests showed that this assay can accurately and reproducibly determine increases in adenosine in fermentation broth without any pretreatment to remove proteins and potentially interfering low-molecular-weight molecules. This assay was also applied to high-throughput screening for high adenosine-producing strains. The high selectivity and accuracy of the ADA assay provides rapid and high-throughput analysis of adenosine in large numbers of samples.     

iQPR法处理水对SD鼠生物安全性的研究

iQPR技术处理污水是一项新型尖端的技术,此技术可以成功降低污水乃至受到污染的地下水中的各种污染指标。但是,iQPR技术处理污水尤其是地下水是否存在潜在的生物安全性问题有待于进一步研究。因此,为评估iQPR技术对生物安全性的影响,本研究首先分析了三种不同iQPR法处理水的水质成分;其次系统研究了iQPR水对SD鼠在个体水平、组织水平和病理形态学损伤的研究。研究表明:iQPR处理的水质成分较对照组普通饮用水好,在个体组织水平检测未见异常,尽管其中一组iQPR处理水造成了SD鼠的脾小体增大,但是可能的原因是水处理环节存在微生物污染现象,因此,初步认定此技术未造成SD大鼠的个体损伤。本研究为揭示iQPR处理的水对生物体的安全性评价提供一个理论依据。     

Systems Mapping for Hematopoietic Progenitor Cell Heterogeneity

Cells with the same genotype growing under the same conditions can show different phenotypes, which is known as “population heterogeneity”. The heterogeneity of hematopoietic progenitor cells has an effect on their differentiation potential and lineage choices. However, the genetic mechanisms governing population heterogeneity remain unclear. Here, we present a statistical model for mapping the quantitative trait locus (QTL) that affects hematopoietic cell heterogeneity. This strategy, termed systems mapping, integrates a system of differential equations into the framework for systems mapping, allowing hypotheses regarding the interplay between genetic actions and cell heterogeneity to be tested. A simulation approach based on cell heterogeneity dynamics has been designed to test the statistical properties of the model. This model not only considers the traditional QTLs, but also indicates the methylated QTLs that can illustrate non-genetic individual differences. It has significant implications for probing the molecular, genetic and epigenetic mechanisms of hematopoietic progenitor cell heterogeneity.     

埃兹蛋白:生物学特征及其在肿瘤转移中的作用

埃兹蛋白(ezrin)是埃兹蛋白、根蛋白和膜突蛋白(ezrin-radixin-moesin,ERM)家族成员之一,主要参与上皮细胞中细胞骨架与胞膜之间的连接,具有维持细胞形态和运动、连接黏附分子及调节信号转导等功能。近年来的研究发现,埃兹蛋白在肿瘤细胞中的表达异常,提示其在肿瘤的浸润、转移机制中发挥重要作用。     

人间隙连接蛋白 31 相互作用蛋白Annexin Ⅱ的筛选与证实

间隙连接蛋白 31 (connexin31 , Cx31) 是间隙连接蛋白 (connexin) 家族的一员,目前对于 Cx31 的功能及其调节方式知之甚少 . 采用固相多肽合成的方法合成 Cx31 羧基端一个多肽片段 (250~266) ,经 HPLC 纯化后偶联到匙孔血蓝蛋白,免疫新西兰雄兔后采血检测、并纯化、经蛋白质印迹、细胞免疫荧光染色、免疫沉淀证实得到的抗体为特异性抗 Cx31 的抗体 . 运用制备的抗 Cx31 多克隆抗体免疫沉淀, SDS- 聚丙烯酰胺凝胶电泳 (SDS-PAGE) 分离,蛋白质条带回收,蛋白质胶块酶解, Q-TOF 质谱分析,数据库扫描筛选可能相互作用蛋白,运用抗体 pull-down 实验,筛选到可能相互作用蛋白 annexin Ⅱ,经过免疫共沉淀、细胞免疫共定位等实验证实 annexin Ⅱ与 Cx31 相互作用 .     

Induction of apoptosis and activation of NF-kappaB by CD95 require different signalling thresholds

Mutations in the death domain of the death receptor CD95 (APO-1/Fas) cause lymphoproliferation and autoimmune disease in both lpr(cg) mice and in patients with autoimmune lymphoproliferative syndrome (ALPS) type Ia. By testing lymphocytes from ALPS type Ia patients, comparing heterozygous with homozygous lpr(cg) mice and coexpressing wild-type and mutant CD95 receptors, we demonstrate that induction of apoptosis requires two wild-type alleles of CD95. By contrast, nuclear factor-kappaB (NF-kappaB) can be fully activated in cells expressing both a mutant and a wild-type CD95 allele, suggesting different thresholds to activate the two signalling pathways. This was confirmed by testing lymphocytes from heterozygous lpr mice, which showed reduced sensitivity to CD95-mediated apoptosis but normal activation of NF-kappaB when compared with wild-type mice. Mutations in CD95 may eliminate the tumour-suppressive function of CD95, at the same time allowing induction of survival or proliferative pathways, which could contribute to the increased risk for lymphoma seen in ALPS type Ia patients.     

Molecular Imaging Reveals a Progressive Pulmonary Inflammation in Lower Airways in Ferrets Infected with 2009 H1N1 Pandemic Influenza Virus

Molecular imaging has gained attention as a possible approach for the study of the progression of inflammation and disease dynamics. Herein we used [¹⁸F]-2-deoxy-2-fluoro-D-glucose ([¹⁸F]-FDG) as a radiotracer for PET imaging coupled with CT (FDG-PET/CT) to gain insight into the spatiotemporal progression of the inflammatory response of ferrets infected with a clinical isolate of a pandemic influenza virus, H1N1 (H1N1pdm). The thoracic regions of mock- and H1N1pdm-infected ferrets were imaged prior to infection and at 1, 2, 3 and 6 days post-infection (DPI). On 1 DPI, FDG-PET/CT imaging revealed areas of consolidation in the right caudal lobe which corresponded with elevated [¹⁸F]-FDG uptake (maximum standardized uptake values (SUVMax), 4.7–7.0). By days 2 and 3, consolidation (CT) and inflammation ([¹⁸F]-FDG) appeared in the left caudal lobe. By 6 DPI, CT images showed extensive areas of patchy ground-glass opacities (GGO) and consolidations with the largest lesions having high SUVMax (6.0–7.6). Viral shedding and replication were detected in most nasal, throat and rectal swabs and nasal turbinates and lungs on 1, 2 and 3 DPI, but not on day 7, respectively. In conclusion, molecular imaging of infected ferrets revealed a progressive consolidation on CT with corresponding [¹⁸F]-FDG uptake. Strong positive correlations were measured between SUVMax and bronchiolitis-related pathologic scoring (Spearman’s ρ = 0.75). Importantly, the extensive areas of patchy GGO and consolidation seen on CT in the ferret model at 6 DPI are similar to that reported for human H1N1pdm infections. In summary, these first molecular imaging studies of lower respiratory infection with H1N1pdm show that FDG-PET can give insight into the spatiotemporal progression of the inflammation in real-time.