P130Cas Src-binding and substrate domains have distinct roles in sustaining focal adhesion disassembly and promoting cell migration

The docking protein p130Cas is a prominent Src substrate found in focal adhesions (FAs) and is implicated in regulating critical aspects of cell motility including FA disassembly and protrusion of the leading edge plasma membrane. To better understand how p130Cas acts to promote these events we examined requirements for established p130Cas signaling motifs including the SH3-binding site of the Src binding domain (SBD) and the tyrosine phosphorylation sites within the substrate domain (SD). Expression of wild type p130Cas in Cas -/- mouse embryo fibroblasts resulted in enhanced cell migration associated with increased leading-edge actin flux, increased rates of FA assembly/disassembly, and uninterrupted FA turnover. Variants lacking either the SD phosphorylation sites or the SBD SH3-binding motif were able to partially restore the migration response, while only a variant lacking both signaling functions was fully defective. Notably, the migration defects associated with p130Cas signaling-deficient variants correlated with longer FA lifetimes resulting from aborted FA disassembly attempts. However the SD mutational variant was fully defective in increasing actin assembly at the protruding leading edge and FA assembly/disassembly rates, indicating that SD phosphorylation is the sole p130Cas signaling function in regulating these processes. Our results provide the first quantitative evidence supporting roles for p130Cas SD tyrosine phosphorylation in promoting both leading edge actin flux and FA turnover during cell migration, while further revealing that the p130Cas SBD has a function in cell migration and sustained FA disassembly that is distinct from its known role of promoting SD tyrosine phosphorylation.     

Transplacental effect of methylcobalamine on the growth of embryonic mouse kidney tissue in organotypic cultivation

Transplacental growth-stimulating effect of the Cbl-coenzyme methylcobalamine (MeCbl) was revealed in organ cultures of embryonic kidney tissue in DBA/2 mice. MeCbl improved the survival of embryonic kidney explants and decreased the incidence of degenerative damage in cultures. The frequency of regeneration development was 30%. Hyperplastic epithelial changes were much more marked after transplacental MeCbl administration (71.8%), than in the control (9.0%).     

Identification and nucleotide polymorphisms in Brassica rapa genes coding cold-shock domain proteins

Full-length BrCSDP2 and BrCSDP4 cold-shock gene sequences of Brassica rapa have been obtained. The isolated genes were shown to belong to a group AtCSP2/AtCSP4 of Arabidopsis thaliana and TsCSDP2/TsCSDP4 of Thellungiella salsuginea genes that encode proteins with a cold-shock domain and two zinc finger motifs. The structure and the allelic variants of these genes have been described and characterized. The identified allelic polymorphism was shown to result both from point substitutions and small indels. Coefficients of total genetic similarity ranged from 1.0 to 0.53. For BrCSDP2 and AtCSDP2, the genetic similarity coefficient was 0.89 and, for BrCSDP4 and AtCSDP4, it was 0.85. The in silico translation of gene sequences has revealed amino acid substitutions in the protein sequence, but no significant correlation between the detected polymorphism and signs of resistance to cold stress were found.     

Serotonin 2B Receptor Antagonism Prevents Heritable Pulmonary Arterial Hypertension

Serotonergic anorexigens are the primary pharmacologic risk factor associated with pulmonary arterial hypertension (PAH), and the resulting PAH is clinically indistinguishable from the heritable form of disease, associated with BMPR2 mutations. Both BMPR2 mutation and agonists to the serotonin receptor HTR2B have been shown to cause activation of SRC tyrosine kinase; conversely, antagonists to HTR2B inhibit SRC trafficking and downstream function. To test the hypothesis that a HTR2B antagonist can prevent BMRP2 mutation induced PAH by restricting aberrant SRC trafficking and downstream activity, we exposed BMPR2 mutant mice, which spontaneously develop PAH, to a HTR2B antagonist, SB204741, to block the SRC activation caused by BMPR2 mutation. SB204741 prevented the development of PAH in BMPR2 mutant mice, reduced recruitment of inflammatory cells to their lungs, and reduced muscularization of their blood vessels. By atomic force microscopy, we determined that BMPR2 mutant mice normally had a doubling of vessel stiffness, which was substantially normalized by HTR2B inhibition. SB204741 reduced SRC phosphorylation and downstream activity in BMPR2 mutant mice. Gene expression arrays indicate that the primary changes were in cytoskeletal and muscle contractility genes. These results were confirmed by gel contraction assays showing that HTR2B inhibition nearly normalizes the 400% increase in gel contraction normally seen in BMPR2 mutant smooth muscle cells. Heritable PAH results from increased SRC activation, cellular contraction, and vascular resistance, but antagonism of HTR2B prevents SRC phosphorylation, downstream activity, and PAH in BMPR2 mutant mice.     

Molecular characterisation and genetic mapping of candidate genes for qualitative disease resistance in perennial ryegrass (Lolium perenne L.)

Background  

Qualitative pathogen resistance in both dicotyledenous and monocotyledonous plants has been attributed to the action of resistance (R) genes, including those encoding nucleotide binding site – leucine rich repeat (NBS-LRR) proteins and receptor-like kinase enzymes. This study describes the large-scale isolation and characterisation of candidate R genes from perennial ryegrass. The analysis was based on the availability of an expressed sequence tag (EST) resource and a functionally-integrated bioinformatics database.     

高中生网络使用状况与网络成瘾知识调查与分析

目的：对高中生网络使用状况和网络成瘾知识进行调查,为学校网络教育提供依据。方法：采用问卷调查法,对北京某高中高二学生进行整群抽样。结果：学生上网目的主要依次为游戏、听歌看电影、获取信息、学习,对网络成瘾有关的知识知晓率较高,但也存在一些网络成瘾的不良行为。结论：调查对象对网络使用较为健康,网络教育尚需加强。     

Alteration of active and passive Wistar rats adaptive behavior in water-immersion model of depression

Animals with active and passive behavioral strategy were selected among male Wistar rats by testing them in the T-maze. Using the results of rats testing in T-maze index of behavioral activity and index of behavioral passivity were calculated. The development of post-stress psychopathology and ils correlation with initial behavioral strategy were studied under water-immersion stress conditions consiclering individual alteration of adaptive behavior. Two paradigms were used--one time trial (stress) and two times trial (stress-restress). It was found that active and passive rats being subjected to inescapable aversive exposure develop different types of post-stress depression and only passive animals show the signs of posttraumatic stress disorder.     

Monoclonal antibodies to Legionella pneumophila cytolysin

The fusion of spleen cells, taken from BALB/c mice immunized with the purified preparation of L. pneumophila cytolysin, with cells Sp2/0 and NP has been carried out. As a result, hybridoma cells producing IgG1, IgG3 and IgM antibodies to this protein have been obtained. All monoclonal antibodies (McAb) thus obtained react with L. pneumophila strain lysates in the precipitation test, while IgG3 and IgM antibodies react with erythrocyte diagnostic agents prepared from the lysate of L. pneumophila cells in the hemagglutination test. In the Western blot assay, McAb react with the 37 KD protein (cytolysin) and a number of other proteins from L. pneumophila cultures and L. pneumophila cell lysate, but do not react with the species-specific protein with a molecular weight of 29 KD, contained in the outer membrane of L. pneumophila, as well as with other species: L. bozemanii, L. dumoffii, L. longbeachae, L. micdadei. The possibility of using these McAb conjugated with FITC and peroxidase for the rapid diagnosis of Legionella infection is shown.