Applications of medium-voltage STEM for the 3-D study of organelles within very thick sections

Scanning transmission electron microscopy at 300 kV enables the visualization of nucleolar silver-stained structures within thick sections (3–8 μm) of Epon-embedded cells at high tilt angles (–50°; + 50°). Thick sections coated with gold particles were used to determine the best conditions for obtaining images with high contrast and good resolution. For a 6-μm-thick section the values of thinning and shrinkage under the beam are 35 to 10%, respectively. At the electron density used in these experiments (100e^—/Å₂/s) it is estimated that these modifications of the section stabilized in less than 10 min. The broadening of the beam through the section was measured and calculations indicated that the subsequent resolution reached 100 nm for objects localized near the lower side of 4-μm-thick sections with a spot-size of 5·6 nm. Comparing the same biological samples, viewed alternately in CTEM and STEM, demonstrated that images obtained in STEM have a better resolution and contrast for sections thicker than 3 μm. Therefore, the visualization of densely stained structures, observed through very thick sections in the STEM mode, will be very useful in the near future for microtomographic reconstruction of cellular organelles.     

Intranucleolar localization of DNA topoisomerase IIalpha is a distinctive feature of necrotic, but not of apoptotic, Jurkat T-cells

Two distinct types of cell death have been described: apoptosis and necrosis. However, it is becoming increasingly clear that the differences between these two types are far less numerous than initially thought. Morphological analyses might provide important information to distinguish apoptotic from necrotic samples. We recently reported that in necrotic, but not apoptotic, HL-60 human myeloid leukaemia cells, the nuclear protein topoisomerase IIalpha concentrated in nucleoli. In order to ascertain whether or not this phenomenon was restricted to a peculiar cell type or could be detected also in cells of lymphoid lineage, we performed an investigation aimed at defining the localization of topoisomerase IIalpha in apoptotic and necrotic Jurkat human T lymphoblastoid cells. Immunofluorescence staining demonstrated that topoisomerase IIalpha was excluded from the condensed chromatin of apoptotic cells, whereas in necrotic cells it was localized in discrete nuclear dots. Immuno-electron microscopy analysis showed that topoisomerase IIalpha was undetectable in nucleoli of normal and apoptotic cells, whereas it was present in the nucleolus of necrotic cells irrespectively of the type of inducer used (ethanol, H(2)O(2), HgCl(2)). Taken together, our findings identify topoisomerase IIalpha as a potential morphological marker useful to discriminate between apoptotic and necrotic cells.     

Non-Coding RNA-Driven Regulation of rRNA Biogenesis

Ribosomal RNA (rRNA) biogenesis takes place in the nucleolus, the most prominent condensate of the eukaryotic nucleus. The proper assembly and integrity of the nucleolus reflects the accurate synthesis and processing of rRNAs which in turn, as major components of ribosomes, ensure the uninterrupted flow of the genetic information during translation. Therefore, the abundant production of rRNAs in a precisely functional nucleolus is of outmost importance for the cell viability and requires the concerted action of essential enzymes, associated factors and epigenetic marks. The coordination and regulation of such an elaborate process depends on not only protein factors, but also on numerous regulatory non-coding RNAs (ncRNAs). Herein, we focus on RNA-mediated mechanisms that control the synthesis, processing and modification of rRNAs in mammals. We highlight the significance of regulatory ncRNAs in rRNA biogenesis and the maintenance of the nucleolar morphology, as well as their role in human diseases and as novel druggable molecular targets.     

微波预处理水酶法提取杨梅核仁油的研究

以杨梅核仁为原料,先采用微波处理,再用纤维素酶和蛋白酶水解,通过正交试验考察制备杨梅核仁油的最佳条件。结果表明,微波处理的效果优于蒸汽处理和热水浸泡两种预处理方式;影响纤维素酶和蛋白酶酶解杨梅核仁提取杨梅核仁油的主次因素为：复合酶用量＞酶解温度＞酶解时间＞料液比;最佳条件为：复合酶用量3.5%、酶解时间90min、酶解温度50℃、料液比1:3,在此条件下总油提取率为53.79%;杨梅核仁油的理化指标符合食用油脂标准;其脂肪酸中不饱和脂肪酸占87.43%,特别是亚油酸含量达46.14%,杨梅核仁油具有较高的营养价值。     

凸随机合作对策的核仁

将合作对策的核仁推广,建立了凸随机合作对策的核仁概念,并探讨了凸随机合作对策的核仁的存在性及与核仁稳定集的关系.     

The lysine-rich C-terminal repeats of the centromere-binding factor 5 (Cbf5) of Kluyveromyces lactis are not essential for function

The gene coding for the centromere-binding factor 5 (CBF5) of Kluyveromyces lactis has been isolated by hybridization of a Saccharomyces cerevisiae CBF5 DNA probe to a K. lactis library. The amino acid sequence of KlCbf5 is highly homologous, 88% identity, to ScCbf5, but also to the rat protein Nap57 (64% identity). The main difference between both yeast proteins and the rat protein is the presence of a lysine-rich domain with KKE/D repeats in the C-terminal part of the protein. These repeats are thought to be involved in binding of the protein to microtubules. Deletion of the KKE/D domain in KlCbf5 however, has no discernible effect on growth on rich medium, sensitivity to the microtubule-destabilizing drug benomyl or segregation of a reporter plasmid. On the other hand, insertion of two leucine residues adjacent to the KKE domain increases the loss rate of a reporter plasmid. In both yeasts complementation of a lethal CBF5 disruption with the heterologous gene results in a slight increase in benomyl sensitivity. A possible role of CBF5 in chromosome segregation will be discussed. © 1998 John Wiley & Sons, Ltd.     

分区核仁法分摊多区域互联输电网固定成本

探讨了电力市场环境下多区域互联电网固定成本的分摊问题。给出固定成本分摊的合作博弈模型,采用分区核仁算法求解。将多区域互联电网拆分为各个区域电网,固定成本根据区域间等值双边交易的比例分摊,再用核仁算法分区求得区域内部各用户承担的固定成本。此方法减小了将多区域互联电网作为一个整体求核仁解的计算量,并且保持了合作博弈模型原本给出的经济信号。IEEE14节点3区域系统的仿真算例验证了该方法的有效性。     

基于合作博弈成本分摊理论的电力系统备用容量分配的研究

为探求公平有效的电力系统备用容量分配方法,以某大区域实际电力负荷和机组参数数据为例,运用基于合作博弈的成本分摊理论进行分析计算,并指明各算法的不同适用范围。算例结果表明核仁分摊法和Shapley值法分配备用容量较现有比例分配法更具公平性、合理性。     

市场机制下机组启动费用的分摊研究

尝试运用基于合作博弈的成本分摊理论来分摊机组启动费用。研究了机组启动费用分摊的核的存在性问题,并指出该分摊不一定是凸博弈。分析了核仁分摊法和Shapley值分摊法的特点和适用范围,对比了两者的优缺点。此外,对机组空载费用的分摊问题也进行了有益的探索。文中,提出采用分时段分摊法和整体分摊法两种方法来分摊多时段的机组启动费用,并对比研究了两者的分摊结果。所提出的分摊方法与现有方法相比,在分摊合理性和公平性方面存在理论上的优势。数值算例结果表明,提出的分摊方法是公平、有效的。