Measuring low levels of protein aggregation by sedimentation velocity

The required performance of an analytical method depends on the purpose for which it will be used. As a methodology matures, it may find new application, and the performance demands placed on the method can increase. Sedimentation velocity analytical ultracentrifugation (SV-AUC) has a long and distinguished history with important contributions to molecular biology. Now the technique is transitioning into industrial settings, and among them, SV-AUC is now used to quantify the amount of protein aggregation in biopharmaceutical protein products, often at levels less than 1% of the total protein mass. In this paper, we review recent advances to SV methodology which have been shown to improve quantitation of protein aggregation. Then we discuss the performance of the SV method in its current state, with emphasis on the precision and quantitation limit of the method, in the context of existing industrial guidance on analytical method performance targets for quantitative methods.     

A parallel study of different array-CGH platforms in a set of Spanish patients with developmental delay and intellectual disability

Developmental delay and intellectual disability, which occur in 1–3% of the population, account for a large number of the cases regularly seen in genetic units. Chromosomal microarray analysis has been shown to be a valuable clinical diagnostic assay and it should be the first-tier clinical diagnostic test for individuals with these conditions. However and due to several difficulties such as the platform resolution, the cost, and the inexperience with genomic data bases, the implementation of this test in many cytogenetic laboratories has been delayed. In an attempt to provide more insights of the benefits derived by using the chromosomal microarray analysis, this study presents the experience of two clinical centers using three different microarray platforms. The results obtained using a custom microarray (KaryoArray®) and two different commercial medium- and high-resolution whole-genome oligonucleotide microarrays have been compared. An overall diagnostic yield of around 15% has been obtained. However, the custom microarray platform has been shown to be more convenient for a clinical setting, since it allows the detection of more pathogenic copy number variants and less common variants.     

A conserved asparagine residue in transmembrane segment 1 (TM1) of serotonin transporter dictates chloride-coupled neurotransmitter transport

Na(+)- and Cl(-)-dependent uptake of neurotransmitters via transporters of the SLC6 family, including the human serotonin transporter (SLC6A4), is critical for efficient synaptic transmission. Although residues in the human serotonin transporter involved in direct Cl(-) coordination of human serotonin transport have been identified, the role of Cl(-) in the transport mechanism remains unclear. Through a combination of mutagenesis, chemical modification, substrate and charge flux measurements, and molecular modeling studies, we reveal an unexpected role for the highly conserved transmembrane segment 1 residue Asn-101 in coupling Cl(-) binding to concentrative neurotransmitter uptake.     

捕食线虫真菌对松材线虫捕食率测定的新方法

本文依据捕食线虫真菌的捕食特性以及松材线虫的生长习性,对松材线虫进行了定时,定量的测定试验,试图通过培养后线虫的种群数量变化,来摸索定量测定捕食率的方法.在漏斗法基础上进行了相应的改进,成功地摸索出3种方法捕食率测定的新方法.各方法各具优缺点,滤膜镜检法和双层镜检法的数据更精确,条切镜检法和漏斗法所测数值较粗略.因此,可依据实验的侧重点以及实验所需的精密程度,而采用最适宜的方法进行捕食率测定实验.     

Optimized intermolecular potential for nitriles based on Anisotropic United Atoms model

An extension of the anisotropic united atoms intermolecular potential model is proposed for nitriles. The electrostatic part of the intermolecular potential is calculated using atomic charges obtained by a simple Mulliken population analysis. The repulsion-dispersion interaction parameters for methyl and methylene groups are taken from transferable AUA4 literature parameters [Ungerer et al., J. Chem. Phys., 2000, 112, 5499]. Non-bonding Lennard-Jones intermolecular potential parameters are regressed for the carbon and nitrogen atoms of the nitrile group (–C≡N) from experimental vapor-liquid equilibrium data of acetonitrile. Gibbs Ensemble Monte Carlo simulations and experimental data agreement is very good for acetonitrile, and better than previous molecular potential proposed by Hloucha et al. [J. Chem. Phys., 2000, 113, 5401]. The transferability of the resulting potential is then successfully tested, without any further readjustment, to predict vapor-liquid phase equilibrium of propionitrile and n-butyronitrile. Figure Saturated vapour pressure of nitriles calculated in this work by molecular simulation compared to experimental data: a) for acetonitrile and b) for both propionitrile and butyronitrile     

Major plant communities of the Carson Desert (Nevada), North America's coldest and driest desert

This paper describes the vegetation of the Carson Desert (Nevada, USA) based on a phytosociological analysis of its major plant associations, as determined by the Braun-Blanquet method. Diagnostic tables, climatic, edaphic, and biogeographical data were used to establish floristic affinities among identified plant communities and to interpret their distributions in zonal gradients. Their syntaxonomic positions in the classes Allenrolfeetea occidentalis and Artemisio tridentatae–Juniperetea osteospermae, and in the new class Sarcobatetea vermiculati were also established. Three associations (Atriplici confertifoliae–Sarcobatetum baileyi, Atriplici canescentis–Psorothamnetum polydenii, and Suaedo moquinii–Sarcobatetum vermiculati) are described here for the first time along with the order Sarcobatetalia vermiculati and the alliance Sarcobation vermiculati.     

小鼠胃 Cajal 间质细胞分离与培养实验方法探讨

目的:尝试优化体外培养Balb/c小鼠胃Cajal间质细胞(interstitial cells of Cajal,ICC)的实验方法,为深入探索该细胞的生理病理作用机制提供基础。方法:无菌条件下取出小鼠胃组织,使用酶解法消化分离细胞,将细胞悬液接种于含有SCF(干细胞因子)的M199培养基中培养,并进行传代。倒置显微镜下观察不同时间段细胞生长状态,采用ICC特异性标志物c-Kit(酪氨酸激酶受体)进行免疫荧光鉴定。结果:细胞培养24 h后基本已贴壁,呈梭形或三角形,有短突起;72 h后细胞胞体变大,突起伸长;5 d后,细胞之间通过突起彼此相互连接,开始形成网状结构;传代后细胞依然保持其固有特征。免疫荧光鉴定可见细胞c-Kit抗体荧光染色阳性。结论:使用酶解法成功分离细胞,细胞数量较多但不增殖,传代后可见细胞纯度较好,稳定培养3周以上后细胞形态逐渐发生变化并开始凋亡。     

超声调制生物介质中光子自相关性质研究

本文首次用Monte Carlo方法研究了超声调制生物介质中散射光子的时间自相关性质,讨论了超声参数、介质的散射系数和吸收系数对自相关函数的影响。正常生物组织和病变生物组织的自相关函数有明显的判别,超声调制自相关函数为光学医学诊断提供了一种新参考。     

酸奶中乳酸菌镜检涂片的特殊染色法

报道了酸奶中乳酸菌涂片的特殊染色法-甲苯胺蓝染色法。涂片固定后,用0．2％甲苯胺蓝染色液染色2min,然后,用1％乙酸溶液脱色数秒,即可使涂片中的菌体着奥蓝色,而背景呈现白色或无色,菌体形状十分清晰。即使涂片很厚且不均,也可清晰看到菌体。便于菌体细胞的计数,形态观察及摄影．