Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

Regardless of their tissue of origin, multipotent mesenchymal stromal cells (MSCs) are commonly expanded in vitro for several population doublings to achieve a sufficient number of cells for therapy. Prolonged MSC expansion has been shown to result in phenotypical, morphological and gene expression changes in MSCs, which ultimately lead to the state of senescence. The presence of senescent cells in therapeutic MSC batches is undesirable because it reduces their viability, differentiation potential and trophic capabilities. Additionally, senescent cells acquire senescence-activated secretory phenotype, which may not only induce apoptosis in the neighboring host cells following MSC transplantation, but also trigger local inflammatory reactions. This review outlines the current and promising new methodologies for the identification of senescent cells in MSC cultures, with a particular emphasis on non-destructive and label-free methodologies. Technologies allowing identification of individual senescent cells, based on new surface markers, offer potential advantage for targeted senescent cell removal using new-generation senolytic agents, and subsequent production of therapeutic MSC batches fully devoid of senescent cells. Methods or a combination of methods that are non-destructive and label-free, for example, involving cell size and spectroscopic measurements, could be the best way forward because they do not modify the cells of interest, thus maximizing the final output of therapeutic-grade MSC cultures. The further incorporation of machine learning methods has also recently shown promise in facilitating, automating and enhancing the analysis of these measured data.     

THE BIOMPHALARIA HAVANENSIS COMPLEX (GASTROPODA: PLANORBIDAE) IN CUBA: A MORPHOLOGICAL AND GENETIC STUDY

Conchological, anatomical and genetic characteristics werecompared among several populations of the Biomphalaria havanensis complexfrom Cuba and other localities in the Caribbean region. Two morphometricmeasures (height and diameter) distinguished two closely similarmorphs of the shell. The reproductive system also separated thesemorphs, particularly upon comparison of sizes of the penis sheathand the preputium. The two morphological groups differed in9 to 12 fixed alleles according to population. Further allozymicdata analyzed in the present work confirmed this difference.The results strongly support the assumption that the B. havanensis complexincludes two distinct species: B. havanensis, for which thetype locality is the swampy area near the vicinity of Havana,Cuba and Biomphalaria sp. a species which commonly occurs inCuba and in Dominican Republic. The taxonomic identity of Biomphalariasp. is discussed. (Received 28 January 2000; accepted 15 August 2000)     

里氏木霉与黑曲霉混合发酵产纤维素酶及其水解特性

研究了利用里氏木霉和黑曲霉混合培养产纤维素酶,以黑曲霉孢子悬浮液的不同活化浓度及不同的活化时间来寻找2个菌种发挥最大协同作用的结合点以及所产纤维素酶的水解特性。以里氏木霉单一培养和黑曲霉单一培养为参照进行对比研究。底物为农林废弃物之一的玉米秸秆,经过蒸气爆破预处理后,用作产酶C源。结果表明:黑曲霉孢子悬浮液活化浓度为10个/mL,活化时间为12 h时,滤纸酶比酶活最高,达3.32 U/mL,高于里氏木霉单一培养的2.25 U/mL,β-葡萄糖苷酶比酶活达1.32 U/mL,高于里氏木霉单一培养的0.57 U/mL。为进一步验证混合菌产纤维素酶的水解效果,利用混合菌产纤维酶的酶液及里氏木霉产纤维素酶的酶液进行酶水解实验,当酶用量为20 U/g绝干纤维素,底物质量浓度为100 g/L条件下水解48 h,混合菌所产酶液酶解得率达70.00%,高于里氏木霉所产酶液的酶解得率63.05%。实验表明里氏木霉与黑曲霉混合培养产酶是可行的,并优于单一菌种培养。     

Differentiating aquatic plant communities in a eutrophic river using hyperspectral and multispectral remote sensing

1. This study evaluates the efficacy of remote sensing technology to monitor species composition, areal extent and density of aquatic plants (macrophytes and filamentous algae) in impoundments where their presence may violate water‐quality standards. 2. Multispectral satellite (IKONOS) images and more than 500 in situ hyperspectral samples were acquired to map aquatic plant distributions. By analyzing field measurements, we created a library of hyperspectral signatures for a variety of aquatic plant species, associations and densities. We also used three vegetation indices. Normalized Difference Vegetation Index (NDVI), near‐infrared (NIR)‐Green Angle Index (NGAI) and normalized water absorption depth (D_H)_, at wavelengths 554, 680, 820 and 977 nm to differentiate among aquatic plant species composition, areal density and thickness in cases where hyperspectral analysis yielded potentially ambiguous interpretations. 3. We compared the NDVI derived from IKONOS imagery with the in situ, hyperspectral‐derived NDVI. The IKONOS‐based images were also compared to data obtained through routine visual observations. Our results confirmed that aquatic species composition alters spectral signatures and affects the accuracy of remote sensing of aquatic plant density. The results also demonstrated that the NGAI has apparent advantages in estimating density over the NDVI and the D_H. 4. In the feature space of the three indices, 3D scatter plot analysis revealed that hyperspectral data can differentiate several aquatic plant associations. High‐resolution multispectral imagery provided useful information to distinguish among biophysical aquatic plant characteristics. Classification analysis indicated that using satellite imagery to assess Lemna coverage yielded an overall agreement of 79% with visual observations and >90% agreement for the densest aquatic plant coverages. 5. Interpretation of biophysical parameters derived from high‐resolution satellite or airborne imagery should prove to be a valuable approach for assessing the effectiveness of management practices for controlling aquatic plant growth in inland waters, as well as for routine monitoring of aquatic plants in lakes and suitable lentic environments.     

家族性高胆固醇血症低密度脂蛋白受体第13外显子突变分析

目的:探讨本课题组收集家族性高胆固醇血症(FH)患者中存在低密度脂蛋白受体(LDLR)第13外显子(E13)基因突变患者临床生化和心血管系统损害特点.方法:对9例临床诊断为FH、基因检测到LDLR基因E13突变的患者进行回顾性分析.结果:(1)临床诊断FH纯合子患者7名,其总胆固醇(TC)水平15.12～26.14 mmol/L,杂合子患者2名,TC水平11.30～11.75 mrnol/L.(2)均可见不同程度黄色瘤;(3)FH纯合子3例心电图出现ST-T改变;4例儿童和1例青年患者出现瓣膜损害,冠脉血流储备(CVFR)减低;杂合子心电图检查均正常,1例出现瓣膜损害,CVFR均正常.(4)核苷酸序列分析证实:9例E13突变患者中,A606T纯舍突变3名;D601Y纯合突变2名;A606T+、W462X和A606T+D601Y复合杂合突变各1名;A606T和D601Y杂合突变各1名.结论:FH严重损害患儿心血管系统和皮肤,LDLR基因E13出现的A606T和D601Y突变可能成为中国FH人群的高频突变位点.     

Comparative transcriptome analysis of developmental stages of the Limonium bicolor leaf generates insights into salt gland differentiation

With the expansion of saline land worldwide, it is essential to establish a model halophyte to study the salt‐tolerance mechanism. The salt glands in the epidermis of Limonium bicolor (a recretohalophyte) play a pivotal role in salt tolerance by secreting excess salts from tissues. Despite the importance of salt secretion, nothing is known about the molecular mechanisms of salt gland development. In this study, we applied RNA sequencing to profile early leaf development using five distinct developmental stages, which were quantified by successive collections of the first true leaves of L. bicolor with precise spatial and temporal resolution. Specific gene expression patterns were identified for each developmental stage. In particular, we found that genes controlling salt gland differentiation in L. bicolor may evolve in a trichome formation, which was also confirmed by mutants with increased salt gland densities. Genes involved in the special ultrastructure of salt glands were also elucidated. Twenty‐six genes were proposed to participate in salt gland differentiation. Our dataset sheds light on the molecular processes underpinning salt gland development and thus represents a first step towards the bioengineering of active salt‐secretion capacity in crops.     

发酵抑制物对絮凝酵母戊糖发酵的影响

将絮凝剂加入酵母溶液中,使酵母絮凝成颗粒以此作为固定化酵母进行戊糖发酵。研究了常见发酵抑制物(甲酸、乙酸、糠醛和乳酸等)对絮凝酵母发酵木糖的影响。结果表明:在60.0g/L木糖发酵液中,经过24h发酵,木糖利用率达94.6%,当分别添加抑制物甲酸、乙酸、糠醛、乙醇和乳酸时,聚氧乙烯絮凝酵母分别对其的耐受浓度为0.5、0.5、1.0、30.0和8.0g/L。当抑制物添加量超过各自的耐受浓度后,对絮凝酵母发酵会产生明显的抑制作用。     

Multiple determinants influence root colonization and induction of induced systemic resistance by Pseudomonas chlororaphis O6

Colonization of the roots of tobacco by Pseudomonas chlororaphis O6 induces systemic resistance to the soft-rot pathogen, Erwinia carotovora ssp. carotovara SCC1. A screen of the transposon mutants of P. chlororaphis O6 showed mutants with about a fivefold reduction in ability to induce systemic resistance to the soft-rot disease. These mutations disrupted genes involved in diverse functions: a methyl-accepting chemotaxis protein, biosynthesis of purines, phospholipase C, transport of branched-chain amino acids and an ABC transporter. Additional mutations were detected in the intergenic spacer regions between genes encoding a GGDEF protein and fumarate dehydratase, and in genes of unknown function. The mutants in the ABC transporters did not display reduced root colonization. However, the other mutants had up to 100-fold reduced colonization levels. Generally the production of metabolites important for interactions in the rhizosphere, phenazines and siderophores, was not altered by the mutations. A reduced induction of systemic resistance by a purine biosynthesis mutant with a disrupted purM gene correlated with poor growth rate, lesser production of phenazines and siderophore and low levels of root colonization. These studies showed that multiple determinants are involved in the induction of systemic resistance, with there being a requirement for strong root colonization.     

Information‐theoretic model selection affects home‐range estimation and habitat preference inference: a case study of male Reeves’s Pheasants Syrmaticus reevesii

Reeves’s Pheasant Syrmaticus reevesii is a vulnerable forest bird inhabiting broadleaved habitats dominated by oaks Quercus spp. in central China. Identifying home‐ranges and habitat associations is important for understanding the biology of this species and developing effective management and conservation plans. We used information‐theoretic criteria to evaluate the relative performance of four parametric (exponential power, one‐mode bivariate normal, two‐mode bivariate normal and two‐mode bivariate circle) and two non‐parametric models (adaptive and fixed kernel) for estimating home‐ranges and habitat associations of Reeves’s Pheasants. For parametric models, Akaike’s information criterion (AIC_c) and the likelihood cross‐validation criterion (CVC) were relatively consistent in ranking the bivariate exponential power model the least acceptable, whereas the two‐mode bivariate models performed better. The CVC suggested that kernel models, particularly the adaptive kernel, performed best among all six models evaluated. The average core area and 95% contour area based on the model with greatest support were 6.1 and 54.9 ha, respectively, and were larger than those estimated from other models. The discrepancy in estimates between models with highest and the lowest support decreased as the contour size increased; however, home‐range shapes differed between models. Minimum convex polygons that removed 5% of extreme data points (MCP95) were roughly half the size of home‐ranges based on kernel models. Estimates of home‐range and model evaluation were not affected by sample size (> 50 observations for each bird). Inference about habitat preference based on composition analysis and home‐range overlap varied between models. That with strongest support suggested that Reeves’s Pheasants selected mature fir and mixed forest, avoided farmland, and had mean among‐individual home‐range overlaps of 20%. We recommend non‐parametric methods, particularly the adaptive kernel method, for estimating home‐ranges and core areas for species with complex multi‐polar habitat preferences in heterogeneous environments with large habitat patches. However, we caution against the traditional convenience of using a single model to estimate home‐ranges and recommend exploration of multiple models for describing and understanding the ecological processes underlying space use and habitat associations.     

研究生在基础与临床相结合科研项目中培养方法的几点思考

随着我国科研创新的开展与推进,学生进入实际科研项目已逐渐成为研究生培养的重要方式。基础与,临床相结合的科研项目立论均来自于临床问题,把在患者身上无法实现的方法通过基础实验的手段加以实施和验证,从而达到发现和揭示病因与发病机制的目的,结果甚或对临床产生指导意义。首都医科大学病理与病理生理学系研究生培养点结合这类研究项目的推进,在规范的方法和流程中引导和培养研究生从临床现象中设问、从海量数据中分析和论证的能力,并通过这些训练,使研究生为进一步深入研究奠定基础,共同为转化医学研究做出贡献。