Active genes are sensitive to deoxyribonuclease I during metaphase

The active exogenous murine leukemia virus sequences of mouse cells growing in culture are preferentially digested by deoxyribonuclease I in metaphase chromosomes. As determined by nuclear nick translation, all of the gene sequences of these cells active during interphase are in a deoxyribonuclease I-sensitive conformation during metaphase. This method of nick translation can therefore be used to label chromosomes in situ in order to visualize the active regions of the genome.     

Hyperphenylalaninemia: disaggregation of brain polyribosomes in young rats

A single injection of l-phenzylalanine in 7-day-old rats produtced disaggregation of brain polyribosomties and inhibition of in vitro protein synthesis in cell-free systems prepared from brain. Liver polyribosomes and in vitro protein synthesis in hepatic systems were not affected. In 4-week-old rats these effects on brain protein synthesis did not occur.     

Dopamine: mediator of brain polysome disaggregation after L-dopa

The disaggregation of brain polysomes which is produced by giving large doses of (L)-dopa to rats is not reproduced by administering its metabolite, 3-O-methyldopa, by giving D-dopa, which also depletes the brain of S-adenosylmethionine but is not converted to catecholamines, or by giving the L-dopa after a decarboxylase inhibitor. Polysome disaggregation is potentiated by the prior administration of a monoamine oxidase inhibitor, indicating that formation of a catecholamine is an obligatory requirement. These observations suggest that the mechanism by which L-dopa disaggregates brain polysomes involves its conversion to dopamine within the majority of brain cells.     

Continuity of protein synthesis through cleavage metaphase

Protein synthesis continues without a decline in rate throughout the period of chromosome condensation and of cytokinesis in the first two cleavages of sea urchin embryos. The natural synchrony of the egg populations and the conditions of measurement allowed even a partial inhibition of synthesis to be observed. Our results do not explain the mechanism of inhibition of protein synthesis that occurs at metaphase in cultured mammalian cells, but it shows that such a change in rate is neither universal nor obligatory.     

L-dopa: disaggregation of brain polysomes and elevation of brain tryptophan

One hour after administration of L-dopa (50 to 300 milligrams per kilogram), there is a marked disaggregation of brain polysomes in immature rats. Adult animals show a similar response, but require larger doses of the amino acid (500 milligrams per kilogram). Single doses of L-dopa significantly elevate amounts of tryptophan in the brain; hence their effect on polysomes does not result from the unavailability of this amino acid.     

Chromosomes can congress to the metaphase plate before biorientation

The stable propagation of genetic material during cell division depends on the congression of chromosomes to the spindle equator before the cell initiates anaphase. It is generally assumed that congression requires that chromosomes are connected to the opposite poles of the bipolar spindle ("bioriented"). In mammalian cells, we found that chromosomes can congress before becoming bioriented. By combining the use of reversible chemical inhibitors, live-cell light microscopy, and correlative electron microscopy, we found that monooriented chromosomes could glide toward the spindle equator alongside kinetochore fibers attached to other already bioriented chromosomes. This congression mechanism depended on the kinetochore-associated, plus end-directed microtubule motor CENP-E (kinesin-7).     

改进非参数解集模型在月径流随机模拟中的应用

【目的】研究改进非参数解集模型在陕北地区月径流随机模拟中的适用性,为该区域水资源规划、开发利用和保护等提供决策依据。【方法】采用非参数核密度估计模型模拟年径流,利用改进非参数解集模型将年径流分解为月径流,对月径流进行随机模拟,并探讨改进非参数解集模型中不同正定型矩阵的分解方法。【结果】改进非参数解集模型考虑了分量与总量以及分量与各分量前期值的关系,克服了首尾自相关不一致的问题,对月径流实测序列的均值、均方差、Cv、Cs、最大值、最小值以及一、二阶自相关系数等统计特性保持较好,模拟效果比较理想。交口河站、张村驿站和绥德站S′矩阵为正定矩阵,可采用Cholesky分解法进行分解;刘家河站、志丹站、安塞站和杏河站S′矩阵为非正定矩阵,宜采用Schur分解法进行分解。【结论】利用改进非参数解集模型进行月径流的随机模拟是可行的。     

Isolation and zonal fractionation of metaphase chromosomes from human diploid cells

A simple method for isolating large quantities of human metaphase chromosomes has been developed. Fractionation of chromosomes from peripheral lymphocytes on a large scale is accomplished by velocity sedimentation in an A-XII zonal rotor.     

Streptonigrin: effect on the first meiotic metaphase of the mouse egg

Streptonigrin, an antibiotic and antitumor agent, alters the chromosomes of the mouse ovum during meiosis. Agglutination of bivalents or achromatic gaps and breaks occurred in the larger pairs both in vitro and in vivo. This newly detected cytogenetic effect suggests that such agents can gain access to developing mammalian ova and destroy the normal progress of meiosis.     

Interphase and metaphase resolution of different distances within the human dystrophin gene

Fluorescence in situ hybridization makes possible direct visualization of single sequences not only on chromosomes, but within decondensed interphase nuclei, providing a potentially powerful approach for high-resolution (1 Mb and below) gene mapping and the analysis of nuclear organization. Interphase mapping was able to extend the ability to resolve and order sequences up to two orders of magnitude beyond localization on banded or unbanded chromosomes. Sequences within the human dystrophin gene separated by less than 100 kb to 1 Mb were visually resolved at interphase by means of standard microscopy. In contrast, distances in the 1-Mb range could not be ordered on the metaphase chromosome length. Analysis of sequences 100 kb to 1 Mb apart indicates a strong correlation between interphase distance and linear DNA distance, which could facilitate a variety of gene-mapping efforts. Results estimate chromatin condensation up to 1 Mb and indicate a comparable condensation for different cell types prepared by different techniques.     

Requirement of Cks2 for the first metaphase/anaphase transition of mammalian meiosis

We generated mice lacking Cks2, one of two mammalian homologs of the yeast Cdk1-binding proteins, Suc1 and Cks1, and found them to be viable but sterile in both sexes. Sterility is due to failure of both male and female germ cells to progress past the first meiotic metaphase. The chromosomal events up through the end of prophase I are normal in both CKS2-/- males and females, suggesting that the phenotype is due directly to failure to enter anaphase and not a consequence of a checkpoint-mediated metaphase I arrest.     

Protein tyrosine kinase Wee1B is essential for metaphase II exit in mouse oocytes

Waves of cyclin synthesis and degradation regulate the activity of Cdc2 protein kinase during the cell cycle. Cdc2 inactivation by Wee1B-mediated phosphorylation is necessary for arrest of the oocyte at G2-prophase, but it is unclear whether this regulation functions later during the metaphase-to-anaphase transition. We show that reactivation of a Wee1B pathway triggers the decrease in Cdc2 activity during egg activation. When Wee1B is down-regulated, oocytes fail to form a pronucleus in response to Ca(2+) signals. Calcium-calmodulin-dependent kinase II (CaMKII) activates Wee1B, and CaMKII-driven exit from metaphase II is inhibited by Wee1B down-regulation, demonstrating that exit from metaphase requires not only a proteolytic degradation of cyclin B but also the inhibitory phosphorylation of Cdc2 by Wee1B.     

Induction of metaphase arrest in cleaving Xenopus embryos by the protein kinase p90Rsk

Before fertilization, vertebrate eggs are arrested in metaphase of meiosis II by cytostatic factor (CSF), an activity that requires activation of the mitogen-activated protein kinase (MAPK) pathway. To investigate whether CSF arrest is mediated by the protein kinase p90Rsk, which is phosphorylated and activated by MAPK, a constitutively activated (CA) form of Rsk was expressed in Xenopus embryos. Expression of CA Rsk resulted in cleavage arrest, and cytological analysis showed that arrested blastomeres were in M phase with prominent spindles characteristic of meiotic metaphase. Thus, Rsk appears to be the mediator of MAPK-dependent CSF arrest in vertebrate unfertilized eggs.     

Fat changes during adolescence

Lower thoracic fat, as measured on serial chest plates of 259 children in Ohio, increased in girls, between ages 6.5 and 14.5 years and, in boys, between ages 6.5 and 11.5 years. No evidence for a marked loss of "baby" fat in adolescence or for "waves" of fattening around the time of puberty was found.     

Enzyme dynamics during catalysis

Internal protein dynamics are intimately connected to enzymatic catalysis. However, enzyme motions linked to substrate turnover remain largely unknown. We have studied dynamics of an enzyme during catalysis at atomic resolution using nuclear magnetic resonance relaxation methods. During catalytic action of the enzyme cyclophilin A, we detect conformational fluctuations of the active site that occur on a time scale of hundreds of microseconds. The rates of conformational dynamics of the enzyme strongly correlate with the microscopic rates of substrate turnover. The present results, together with available structural data, allow a prediction of the reaction trajectory.