Total syntheses in solution of TOAC-labelled alamethicin F50/5 analogues

Total syntheses in solution of a set of four selected analogues of the 19-mer component F50/5 of alamethicin, the most extensively studied among the channel-former peptaibol antibiotics, are planned and reported. All analogues bear three Glu(OMe) residues, replacing the Gln residues at positions 7, 18, and 19 of the naturally occurring compound. Three analogues are mono-labelled with the free-radical-containing amino acid residue TOAC at the strategic positions 1, 8, or 16. The fourth analogue is bis-labelled with the same EPR-active residue at both positions 1 and 16. In the native sequence, all of the positions where TOAC replacements have been introduced are characterized by residues of Aib, the prototype of the class of helicogenic C(alpha)-tetrasubstituted alpha-amino acids. All of the TOAC analogues synthesized exhibit significant membrane-modifying properties.     

Urea/thiourea catalyzed, solvent-free synthesis of 5-arylidenethiazolidine-2,4-diones and 5-arylidene-2-thioxothiazolidin-4-ones

An efficient and organo-catalyzed method has been developed for the synthesis of 5-arylidenethiazolidine-2,4-diones and 5-arylidene-2-thioxothiazolidin-4-ones via Knoevenagel condensation of arylaldehydes 1 and 2,4-thiazolidinedione 2a/2-thioxothiazolidin-4-one 2b under mild conditions. Urea-adduct 4 and azomethine 5 also afford arylidene-products 3 by reacting with 2a-b via addition-elimination reaction. This protocol has the features of use of inexpensive, ecofriendly readily available, effective catalyst system viz. urea/thiourea, avoidance of volatile solvents, excellent yield and simple work-up procedure.     

Vanadium mediated apoptosis and cell cycle arrest in MCF7 cell line

Vanadium is a metal widely distributed in the environment. It is also a dietary micronutrient. It has shown insulin mimetic and chemopreventive properties and has been considered as an important pharmacological agent. In this study, we evaluated the apoptogenic role of vanadium on human breast cancer cell line MCF7. Exposure of MCF7 cells to vanadium led to the induction of apoptosis in a dose-dependent manner. Percentage of apoptosis was maximum (42.5%) at the highest non-toxic dose (250 microM). It was found that vanadium treatment brought about a prominent chromatin condensation, cell cycle arrest leading to apoptosis. These apoptosis based assays demonstrate that vanadium has the potential to be developed into an anti-cancer drug in the near future.     

Chromatin condensation of ovarian nurse and follicle cells is regulated independently from DNA fragmentation during Drosophila late oogenesis

Programmed cell death constitutes a common fundamental incident occurring during oogenesis in a variety of different organisms. In Drosophila melanogaster, it plays a significant role in the maturation process of the egg chamber. In the present study, we have used an in vitro development system for studying the effects of inducers and inhibitors of programmed cell death during the late stages of oogenesis. Treatment of the developing egg chambers with two widely used inducers of cell death, etoposide and staurosporine, blocks further development and induces chromatin condensation but not DNA fragmentation in nurse and follicle cells, as revealed by propidium iodide staining and terminal transferase-mediated dUTP nick-end labeling assay. Moreover, incubation of the developing egg chambers with the caspase-3 inhibitor Z-DEVD-FMK significantly delays development, prevents DNA fragmentation, but does not affect chromatin condensation. The above results demonstrate, for the first time, that chromatin condensation in Drosophila ovarian nurse and follicle cells is a caspase-3-like independent process and is regulated independently from DNA fragmentation.     

Integrin signaling and cell spreading alterations by rottlerin treatment of chick limb bud mesenchymal cells

Endochondral skeletal development begins with the formation of a cartilaginous template where mesenchymal cells aggregate and increase in density prior to their overt differentiation into chondrocytes. Prechondrogenic condensation, in which mesenchymal cells aggregate, requires cell migration and proliferation. However, the molecular mechanisms promoting this aggregation remain to be elucidated. Here, we report that rottlerin suppresses migration and cell surface expression of integrin β1 in chondrogenic progenitors. Perturbation of integrin β1 function using an anti-integrin β1 blocking antibody suppressed the migration of wing bud mesenchymal cells. Furthermore, phosphorylation levels of Src and focal adhesion kinase (FAK) were decreased by rottlerin treatment. Cell treatment with PP2, an inhibitor of Src family kinase, or electroporation of FAK specific siRNA, suppressed cell migration in a wound-healing assay. Cells treated with rottlerin showed decreased phosphorylation of Akt, independent of PKCδ inhibition. In addition, an Akt inhibitor suppressed the migration of chick limb bud mesenchymal cells. Taken together, our results point to the novel finding that rottlerin may act as a negative regulator for cell migration, an essential step for prechondrogenic condensation, by regulating integrin β1 signaling at focal adhesion complexes via modulation of Akt activity.     

大兴安岭呼中林区地表死可燃物含水量及其环境梯度分析

森林地表死可燃物含水量与林火行为密切相关,理解森林地表死可燃物含水量及其与环境因子之间的关系对于林火管理有重要意义。本文对大兴安岭呼中林区的不同植被类型内的地表死可燃物含水量,依据1 h、10 h和100 h分类标准进行了对比,结果表明：杨桦林和钻天柳林等2种阔叶林地表死可燃物含水量最高,而其他植被类型比阔叶林内地表死可燃物含水量低,且之间无显著性差异;对兴安落叶松不同林型内的地表死可燃物含水量进行比较,发现越桔兴安落叶松林内地表死可燃物含水量最高,其他林型比越桔兴安落叶松林低,且之间没有显著性差异。此外,利用除趋势典范对应分析对地表死可燃物含水量的环境梯度进行分析。结果表明：第1排序轴反映了坡位与海拔梯度,即热量因素,沿着第1轴从左到右海拔和坡位逐渐降低;第2轴反映了坡向和林分密度梯度,即水分因素。总体来说,坡位与海拔是影响森林地表死可燃物含水量的主要环境因子,二者综合作用表达了该地区森林地表死可燃物含水量的空间生态梯度。     

Chromosome shattering: a mitotic catastrophe due to chromosome condensation failure

Chromosome shattering has been described as a special form of mitotic catastrophe, which occurs in cells with unrepaired DNA damage. The shattered chromosome phenotype was detected after application of a methanol/acetic acid (MAA) fixation protocol routinely used for the preparation of metaphase spreads. The corresponding phenotype in the living cell and the mechanism leading to this mitotic catastrophe have remained speculative so far. In the present study, we used V79 Chinese hamster cells, stably transfected with histone H2BmRFP for live-cell observations, and induced generalized chromosome shattering (GCS) by the synergistic effect of UV irradiation and caffeine posttreatment. We demonstrate that GCS can be derived from abnormal mitotic cells with a parachute-like chromatin configuration (PALCC) consisting of a bulky chromatin mass and extended chromatin fibers that tether centromeres at a remote, yet normally shaped spindle apparatus. This result hints at a chromosome condensation failure, yielding a “shattered” chromosome complement after MAA fixation. Live mitotic cells with PALCCs proceeded to interphase within a period similar to normal mitotic cells but did not divide. Instead they formed cells with highly abnormal nuclear configurations subject to apoptosis after several hours. We propose a factor depletion model where a limited pool of proteins is involved both in DNA repair and chromatin condensation. Chromosome condensation failure occurs when this pool becomes depleted. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. This article has been submitted as a contribution to the festschrift entitled “Uncovering cellular sub-structures by light microscopy” in honour of Professor Cremer’s 65th birthday.     

A novel pocket in 14-3-3? is required to mediate specific complex formation with cdc25C and to inhibit cell cycle progression upon activation of checkpoint pathways

Mitotic progression requires the activity of the dual specificity phosphatase, cdc25C. Cdc25C function is inhibited by complex formation with two 14-3-3 isoforms, 14-3-3? and 14-3-3γ. To understand the molecular basis of specific complex formation between 14-3-3 proteins and their ligands, chimeric 14-3-3 proteins were tested for their ability to form a complex with cdc25C in vivo. Specific complex formation between cdc25C and 14-3-3? in vivo requires a phenylalanine residue at position 135 (F135) in 14-3-3?. Mutation of this residue to the corresponding residue present in other 14-3-3 isoforms (F135V) leads to reduced binding to cdc25C and a decrease in the ability to inhibit cdc25C function in vivo. Similarly, F135V failed to rescue the incomplete S phase and the G2 DNA damage checkpoint defects observed in cells lacking 14-3-3?. A comparative analysis of the 14-3-3 structures present in the database suggested that the F135 in 14-3-3? was required to maintain the integrity of a pocket that might be involved in secondary interactions with cdc25C. These results suggest that the specificity of the 14-3-3 ligand interaction may be dependent on structural motifs present in the individual 14-3-3 isoforms.     

Effects of vegetation cover and precipitation on the process of sediment produced by erosion in a small watershed of loess region

With growing concerns over changes of the living environment and ecological environment, more and more scholars have focused their researches on understanding how vegetation covers and atmospheric conditions respond to soil erosion in watersheds. Former studies show that both the natural factors such as precipitation, vegetation, slope of terrain, soil properties and human activities are the main factors to affect the amount of sediment produced by erosion in the watershed, and there are special conditions of climate and soil that are unique to loess areas for water and soil conservations. Thus the relationships between soil erosion and vegetation and precipitation are very complicated and interesting. As a loess area, the Lüergou watershed with the area of 12.1 km2 lies in west of China. The watershed was a key area of high water and soil erosion forty years ago, but the area of vegetation cover has become larger because of highly effective methods of water and soil conservation. In the factors affecting the amount of sediment produced by erosion in the study area, which is more important for soil erosion: vegetation cover or precipitation? The experts, community has discussed this question for a long time. And the Lüergou watershed has become the natural and ideal test watershed. Based on water observation data, climate data and NOAA/AVHRR NDVI images collected from 1982 to 2000 in the Lüergou watershed of loess areas, analyses of the correlation and multi-variable regression were used to discuss the relationships among the amount of sediment produced by erosion, water indexes, precipitation factors and vegetation cover. The conclusions showed that with the increase of precipitation indexes and the decrease of plant indexes, and the amount of sediment produced by erosion in the study area would become larger. In order to distinguish the influences of erosion due to human activity and natural factors, the paper introduced multi-variable regression method by standardization data to determine the relative contributing ratio to soil erosions in the study area. The conclusions showed that the contributing ratio of vegetation cover and precipitation changes were 45.7% and 54.3%. It was obvious that the influences of precipitation were larger than those of vegetation for the soil erosion in the study area.