Cross-scale quality assessment of a mechanistic cation exchange chromatography model

Cation exchange chromatography (CEX) is an essential part of most monoclonal antibody (mAb) purification platforms. Process characterization and root cause investigation of chromatographic unit operations are performed using scale down models (SDM). SDM chromatography columns typically have the identical bed height as the respective manufacturing-scale, but a significantly reduced inner diameter. While SDMs enable process development demanding less material and time, their comparability to manufacturing-scale can be affected by variability in feed composition, mobile phase and resin properties, or dispersion effects depending on the chromatography system at hand. Mechanistic models can help to close gaps between scales and reduce experimental efforts compared to experimental SDM applications. In this study, a multicomponent steric mass-action (SMA) adsorption model was applied to the scale-up of a CEX polishing step. Based on chromatograms and elution pool data ranging from laboratory- to manufacturing-scale, the proposed modeling workflow enabled early identification of differences between scales, for example, system dispersion effects or ionic capacity variability. A multistage model qualification approach was introduced to measure the model quality and to understand the model's limitations across scales. The experimental SDM and the in silico model were qualified against large-scale data using the identical state of the art equivalence testing procedure. The mechanistic chromatography model avoided limitations of the SDM by capturing effects of bed height, loading density, feed composition, and mobile phase properties. The results demonstrate the applicability of mechanistic chromatography models as a possible alternative to conventional SDM approaches.     

静注氟比洛芬酯预防肺叶手术中全麻患者苏醒期躁动的临床研究

目的:探讨肺叶手术中超前应用氟比洛芬酯对减轻全身麻醉苏醒期躁动发生率的影响。方法:60例肺叶切除术患者随机分为A组和B组,每组30例,两组均常规诱导全麻气管插管,术中瑞芬、丙泊酚、维库溴铵泵住维持麻醉,A组于术前给予氟比洛芬酯1 MG/KG。比较两组躁动得分、苏醒后(10 min、30 min、1 h、2 h、4 h、24 h)六个时间点疼痛评分及不良反应发生率。结果:两组出血时间、拔管时间、麻醉时间差异均无统计学意义(均P0.05);A组患者的躁动得分显著低于B组,差异有统计学意义(P0.05);两组的VAS评分差异有统计学意义(P0.05),不同时间点的VAS评分差异有统计学意义(P0.05),分组与时间之间存在交互作用(P0.05);A组患者的不良反应发生率低于B组,差异有统计学意义(P0.05)。结论:肺叶手术中超前应用氟比洛芬酯具有良好的镇痛效果,可以降低全身麻醉苏醒期躁动的发生率。     

Novel Systems for Dynamically Assessing Insulin Action in Live Cells Reveals Heterogeneity in the Insulin Response

Regulated GLUT4 trafficking is a key action of insulin. Quantitative stepwise analysis of this process provides a powerful tool for pinpointing regulatory nodes that contribute to insulin regulation and insulin resistance. We describe a novel GLUT4 construct and workflow for the streamlined dissection of GLUT4 trafficking; from simple high throughput screens to high resolution analyses of individual vesicles. We reveal single cell heterogeneity in insulin action highlighting the utility of this approach – each cell displayed a unique and highly reproducible insulin response, implying that each cell is hard‐wired to produce a specific output in response to a given stimulus. These data highlight that the response of a cell population to insulin is underpinned by extensive heterogeneity at the single cell level. This heterogeneity is pre‐programmed within each cell and is not the result of intracellular stochastic events.     

五种荒漠灌木群落土壤有机碳垂直分布及其与根系分布的关系

研究土壤有机碳与根系分布之间关系对评估碳储量和固碳潜力具有重要意义,而我国荒漠区灌木群落土壤有机碳与植物根系之间的关系尚不清楚。本研究选取深根性表层型灌木(表层根系生物量最大)梭梭(Haloxylon ammodendro)和中间锦鸡儿(Caragana intermedia)、深根性非表层型(表层根系生物量不是最大)灌木白刺(Nitratia tangutorum)、浅根性表层型灌木红砂(Reaumuria songarica)及浅根性非表层型灌木霸王(Sarcozygium xanthoxylon)为优势种的5种荒漠灌木群落为研究对象,测定100 cm深度内不同层次根系生物量及土壤有机碳含量,探讨灌木群落土壤有机碳的垂直分布格局及其与根系之间的关系。结果显示:(1)0~100 cm土壤有机碳密度与群落0~20 cm根系生物量碳密度显著正相关,与其他层次或累积深度不显著相关;(2)梭梭群落0~100 cm土壤有机碳密度显著低于与其他4个群落(P0.01),其他4个群落无显著差异(P0.05);(3)优势种为表层型的梭梭、中间锦鸡儿和红砂群落土壤有机碳密度与群落及优势种根系变化趋势一致,优势种为浅根性非表层型的霸王群落仅表层40 cm土壤有机碳与优势种灌木根系分布规律一致,优势种为深根性非表层型的白刺群落土壤有机碳与优势种根系分布规律一致。本研究表明,荒漠灌木群落0~100 cm土壤有机碳储量与群落0~20 cm根系生物量显著相关,群落优势种根系分布类型是影响土壤有机碳垂直分布格局的重要因素。     

Conformational analysis of protein structures derived from NMR data

A study is presented of the conformational characteristics of NMR-derived protein structures in the Protein Data Bank compared to X-ray structures. Both ensemble and energy-minimized average structures are analyzed. We have addressed the problem using the methods developed for crystal structures by examining the distribution of ?, Ψ, and χ angles as indicators of global conformational irregularity. All these features in NMR structures occur to varying degrees in multiple conformational states. Some measures of local geometry are very tightly constrained by the methods used to generate the structure, e.g., proline ? angles, α-helix ?, Ψ angles, ω angles, and C_α chirality. The more lightly restrained torsion angles do show increasead clustering as the number of overall experimental observations increases. ?, Ψ, and χ₁ angle conformational heterogeneity is strongly correlated with accessibility but shows additional differences which reflect the differing number of observations possible in NMR for the various side chains (e.g., many for Trp, few for Ser). In general, we find that the core is defined to a notional resolution of 2.0 to 2.3 Å. Of real interest is the behavior of surface residues and in particular the side chains where multiple rotameric states in different structures can vary from 10% to 88%. Later generation structures show a much tighter definition which correlates with increasing use of J-coupling information, stereospecific assignments, and heteronumclear techniques. A suite of programs is being developed to address the special needs of NMR-derived structures which will take into account the existence of increased mobility in solution. © 1993 Wiley-Liss, Inc.     

Scale specific determinants of tree diversity in an old growth temperate forest in China

The major processes generating pattern in plant community composition depend upon the spatial scale and resolution of observation, therefore understanding the role played by spatial scale on species patterns is of major concern. In this study, we investigate how well environmental (topography and soil variables) and spatial variables explain variation in species composition in a 25-ha temperate forest in northeastern China. We used new variation partitioning approaches to discover the spatial scale (using multi-scale spatial PCNM variables) at which environmental heterogeneity and other spatially structured processes influence tree community composition. We also test the effect of changing grain of the study (i.e. quadrat size) on the variation partitioning results. Our results indicate that (1) species composition in the Changbai mixed broadleaf-conifer forest was controlled mainly by spatially structured soil variation at broad scales, while at finer scales most of the explained variation was of a spatial nature, pointing to the importance of biotic processes. (2) These results held at all sampling grains. However, reducing quadrat size progressively reduced both spatially and environmentally explained variance. This probably partly reflects increasing stochasticity in species abundances, and the smaller proportion of quadrats occupied by each species, when quadrat size is reduced. The results suggest that environmental heterogeneity (i.e. niche processes) and other biotic processes such as dispersal work together, but at different spatial scales, to build up diversity patterns.     

氨甲酰胆碱和Impromidine对CFU—S细胞周期的影响

正常情况下处于S期的CFU-S比例低于10％。氨甲酰胆碱(Cach 10~(-13)—10~(-9)mol/L)和Impromidine(Impro 10~(-9)—10~(-4)mol/L)在体外与小鼠骨髓细胞短时培育后,增加了CFU-S对细胞毒剂羟基脲的敏感性。反应最大时,9d和13dCFU-S的减少率分别是32.8和60.6％(Cach)以及38.4和49.5％(Impro)。这种效应可分别被胆碱能N受体阻断剂筒箭毒(10~(-6)mol/L)和组胺H_2受体阻断剂甲氰咪呱(10~(-6)mol/L)所阻断,表明9d和13d CFU-S表面胆碱能N受体和组胺H_2受体的密度或活性存在差别,再次证实了CFU-S是一个不均一的细胞群。     

黑河中游荒漠绿洲过渡带土壤水分与植被空间变异

荒漠绿洲过渡带是绿洲生态系统的重要组成部分,对维持绿洲稳定具有重要作用。过渡带土壤湿度和植被之间的相互关系是了解荒漠绿洲过渡带的重要科学问题,从而开始受到重视。研究是在黑河中游荒漠绿洲过渡带,选择一条1700m×200m的样带（包括3条平行样线）,在对土壤湿度、植被高度、盖度等调查的基础上,应用统计分析和地统计的方法,研究了荒漠绿洲过渡带土壤水分和植被的空间变异性特征。结果表明：在荒漠绿洲过渡带上,0～200cm土层中土壤平均湿度介于1．45％～3．85％,变异系数在27．7％～83．2％;植被盖度介于9％～80％,变异系数为80％。植被盖度、冠幅与0～20、20～40cm两层土壤湿度显著负相关（p〈0．05）,与120～140、140～160cm两层土壤湿度显著正相关（p〈0．01）。土壤水分和植被空间分布在样带上存在明显的空间异质性,表现在小于100m的尺度上随机分布,而在100～3110m的尺度上呈聚集分布格局。     

On statistical inference for the random set generated Cox process with set-marking

Cox point process is a process class for hierarchical modelling of systems of non-interacting points in Rd under environmental heterogeneity which is modelled through a random intensity function. In this work a class of Cox processes is suggested where the random intensity is generated by a random closed set. Such heterogeneity appears for example in forestry where silvicultural treatments like harvesting and site-preparation create geometrical patterns for tree density variation in two different phases. In this paper the second order property, important both in data analysis and in the context of spatial sampling, is derived. The usefulness of the random set generated Cox process is highly increased, if for each point it is observed whether it is included in the random set or not. This additional information is easy and economical to obtain in many cases and is hence of practical value; it leads to marks for the points. The resulting random set marked Cox process is a marked point process where the marks are intensity-dependent. The problem with set-marking is that the marks are not a representative sample from the random set. This paper derives the second order property of the random set marked Cox process and suggests a practical estimation method for area fraction and covariance of the random set and for the point densities within and outside the random set. A simulated example and a forestry example are given.     

Broad-scale habitat use by red-legged partridge <Emphasis Type="Italic">(Alectoris rufa</Emphasis>) in a low-density area in northwestern Spain

We developed broad-scale habitat selection models for the distribution of red-legged partridge Alectoris rufa in a low-density area in northwestern Spain, the Baixa-Limia site of community importance (SCI). The fieldwork consisted of ground surveys in 1 × 1 km squares. For habitat selection analysis, we used a 2 × 2 km grid integrating the information obtained in the 1 × 1 km squares. As predictors we used environmental variables measured on digital 1:50,000 scale cartography using a geographical information system (GIS). The red-legged partridge was scarce in the study area. The logistic regression analysis carried out on data from the squares with probable and confirmed breeding included the area of scrubland and pastureland with a positive sign. Using the breeding index category (BIC) three variables produced a slightly positive response: area of scrubland and pastureland, length of border between scrublands and forests, and length of border between forests and dams. The difficulty for modelling the habitat selection of this species could be due to human activities (hunting, habitat loss, restocking of hunt species), and may have modified their habitat preferences. Furthermore, the occupation of suboptimal habitats would distort the real habitat preferences.