Copper-induced germline apoptosis in Caenorhabditis elegans: The independent roles of DNA damage response signaling and the dependent roles of MAPK cascades

Excess copper is toxic to life. Copper has been shown to induce apoptosis in various cell lines and tissues. However, due to the lack of appropriate gene knockout animal models, data concerning the underlying pathways of copper-induced apoptosis are insufficient, especially with regards to in vivo systems. The nematode Caenorhabditis elegans is a good model to study basic biological processes, including stress responses and apoptosis. In the present study, we investigated copper-induced germline apoptosis in the C. elegans strains carrying mutated alleles of homologs to known mammalian genes that are involved in apoptosis regulation. We show here that exposing C. elegans to copper causes dose- and time-dependent germline apoptosis. The knockout of checkpoint genes hus-1, clk-2, the Bcl-2 homolog ced-9, and the BH3-only domain egl-1 did not prevent cells of the germline from copper-induced apoptosis. The loss-of-function of the tumor suppressor gene, p53/cep-1, caused a significant increase in germline apoptosis with exposure to copper, and the depletion of p53 antagonist ABL1 significantly enhanced apoptosis. The knockout of the caspase gene ced-3 and the Apaf-1 homolog ced-4 abrogated both copper-induced and physiological germline apoptosis. Germline apoptosis stopped increase in the strains lin-45(ku51), mek-2(n1989), mpk-1(ku1) under copper stresses, respectively. Copper-induced apoptosis was blocked in the loss-of-function alleles of both JNK and p38 MAPK cascades excepting pmk-3, one of the three p38 MAPK components. Together, the results of this study suggest that caspase and Apaf-1 are required for copper-induced germline apoptosis while DNA damage response genes are not essential, and that the Raf-MEK-ERK, ASK1/2-MKK7-JNK, ASK1/2-MKK3/6-p38 signaling pathways are indispensable in mediating this apoptotic response.     

Monitoring vegetation recovery after China’s May 2008 Wenchuan earthquake using Landsat TM time-series data: a case study in Mao County

The Wenchuan earthquake (Richter scale 8) on 12 May 2008 in southwestern China caused widespread ecosystem damage in the Longmenshan area. It is important to evaluate natural vegetation recovery processes and provide basic information on ecological aspects of the recovering environment after the earthquake. To circumvent the weather limits of remote sensing in the Wenchuan earthquake-hit areas, and to meet the need for regional observation analyses, three Landsat TM images pre- and post-earthquake in Mao County were used for analysis. Post-earthquake normalized difference vegetation index (NDVI) values were compared to pre-earthquake values with an NDVI-based index differencing method to determine the extent to which the vegetation was damaged in relation to the pre-earthquake pattern, and the rate of recovery was evaluated. The spatial characteristics of vegetation loss and natural recovery patterns were analyzed in relation to elevation, slope and aspect. The results indicated that severely damaged sites occurred mainly in river valleys, within a range of 1,500?C2,500?m elevation and on slopes of 25?C55°. The distance from rivers, rather than the distance from active faults, controls the damage patterns. After 1?year of natural regeneration, 36?% of the destroyed areas showed a decrease in NDVI value, 28.8?% showed very little change, 19.1?% showed an increase, and 16.1?% also increased with a recovery rate greater than 100?%. Moreover, there is a good correlation between recovery rate and both slope and elevation, but recovery patterns in the damaged area are complicated. Our results indicate that natural recovery in this arid valley is a slow process.     

Theoretical Insights into Catalytic Mechanism of Protein Arginine Methyltransferase 1

Protein arginine methyltransferase 1 (PRMT1), the major arginine asymmetric dimethylation enzyme in mammals, is emerging as a potential drug target for cancer and cardiovascular disease. Understanding the catalytic mechanism of PRMT1 will facilitate inhibitor design. However, detailed mechanisms of the methyl transfer process and substrate deprotonation of PRMT1 remain unclear. In this study, we present a theoretical study on PRMT1 catalyzed arginine dimethylation by employing molecular dynamics (MD) simulation and quantum mechanics/molecular mechanics (QM/MM) calculation. Ternary complex models, composed of PRMT1, peptide substrate, and S-adenosyl-methionine (AdoMet) as cofactor, were constructed and verified by 30-ns MD simulation. The snapshots selected from the MD trajectory were applied for the QM/MM calculation. The typical S_N2-favored transition states of the first and second methyl transfers were identified from the potential energy profile. Deprotonation of substrate arginine occurs immediately after methyl transfer, and the carboxylate group of E144 acts as proton acceptor. Furthermore, natural bond orbital analysis and electrostatic potential calculation showed that E144 facilitates the charge redistribution during the reaction and reduces the energy barrier. In this study, we propose the detailed mechanism of PRMT1-catalyzed asymmetric dimethylation, which increases insight on the small-molecule effectors design, and enables further investigations into the physiological function of this family.     

Screening of binding proteins that interact with human Salvador 1 in a human fetal liver cDNA library by the yeast two-hybrid system

Salvador promotes both cell cycle exit and apoptosis through the modulation of both cyclin E and Drosophila inhibitor of apoptosis protein in Drosophila. However, the cellular function of human Salvador (hSav1) is rarely reported. To screen for novel binding proteins that interact with hSav1, the cDNA of hSav1 was cloned into a bait protein plasmid, and positive clones were screened from a human fetal liver cDNA library by the yeast two-hybrid system. hSav1 mRNA was expressed in yeast and there was no self-activation and toxicity in the yeast strain AH109. Twenty proteins were found to interact with hSav1, including HS1 (haematopoietic cell specific protein1)-associated protein X-1 (HAX-1); neural precursor cell expressed, developmentally down-regulated 9, pyruvate kinase, liver and RBC, cytochrome c oxidase subunit Vb, enoyl coenzyme A hydratase short chain 1, and NADH dehydrogenase (ubiquinone) 1 beta subcomplex, demonstrating that the yeast two-hybrid system is an efficient method for investigating protein interactions. Among the identified proteins, there were many mitochondrial proteins, indicating that hSav1 may play a role in mitochondrial function. We also confirmed the interaction of HAX-1 and hSav1 in mammalian cells. This investigation provides functional clues for further exploration of potential apoptosis-related proteins in disease biotherapy.     

The role of GSK3beta in the development of the central nervous system

Glycogen synthase kinase 3β (GSK3β) is a multifunctional serine/threonine kinase.It is particularly abundant in the developing central nervous system (CNS).Since GSK3β has diverse substrates ranging fr...     

粉煤灰基质-草坪砖栽培环境对坪草生长的影响

为了解决停车场、甬路、渠坝与草坪争地的矛盾,增加城市绿地面积,在已研究粉煤灰草坪基质最佳混合比例的基础上,进一步探讨草坪砖作为草坪栽培环境(砖孔内填人粉煤灰混合基质,简称砖孔环境)的肥力效应与生物效应。为了对比分析,特此设置了土孔环境(在土壤中挖取与草坪砖形状和容积相同的孔并填入粉煤灰混合基质)与土灰环境(在粉煤灰混合基质中按照草坪砖孔模式划出同样大小的圈)种植黑麦草作为对照。研究结果表明：坪草N、K、Na、Cu、Zn等含量以砖孔环境最高,土灰环境和土孔环境较低,差异显著,说明砖孔环境能为植物生长提供较多的养分元素;砖孔环境中的坪草含有的Fe、Cu、Zn浓度远高于坪草最佳需求量,今后不再需要补充铁肥、铜肥和锌肥。试验还表明砖孔环境的蒸散率最小,基质含水量最大,基质势最高,持续供水能力强,抗旱效果十分明显。砖孔环境与土孔环境中的草坪草屑累积量差异不显著,但均比土灰环境中的高,且差异显著。夏季高温期砖孔环境中的草坪质量显著高于土孔环境和土灰环境中的草坪质量。     

Optimal Formation Reconfiguration Control of Multiple UCAVs Using Improved Particle Swarm Optimization

Optimal formation reconfiguration control of multiple Uninhabited Combat Air Vehicles （UCAVs） is a complicated global optimum problem. Particle Swarm Optimization （PSO） is a population based stochastic optimization technique inspired by social behaviour of bird flocking or fish schooling. PSO can achieve better results in a faster, cheaper way compared with other bio-inspired computational methods, and there are few parameters to adjust in PSO. In this paper, we propose an improved PSO model for solving the optimal formation reconfiguration control problem for multiple UCAVs. Firstly, the Control Parameterization and Time Diseretization （CPTD） method is designed in detail. Then, the mutation strategy and a special mutation-escape operator are adopted in the improved PSO model to make particles explore the search space more efficiently. The proposed strategy can produce a large speed value dynamically according to the variation of the speed, which makes the algorithm explore the local and global minima thoroughly at the same time. Series experimental results demonstrate the feasibility and effectiveness of the proposed method in solving the optimal formation reconfiguration control problem for multiple UCAVs.     

应用组织芯片技术研究MMP-9和PCNA在宫颈癌组织中的表达及其意义

目的探讨MMP-9和PCNA在宫颈癌组织中的联合表达、相关性及其临床意义。方法采用组织芯片和免疫组化技术检测143例宫颈浸润癌(ICC)、20例癌旁正常宫颈上皮(NCE)中MMP-9和PCNA的表达,统计分析MMP-9、PCNA表达与病理分级、临床分期和淋巴结转移等临床病理因素的关系。结果 MMP-9、PCNA在宫颈癌组织中的阳性率分别为89.5%(128/143)和93.7%(134/143),较正常宫颈组织中的阳性率40.0%(8/20)和15.0%(3/20)显著增高(P=0.000)。MMP-9的表达与病理分级(r=0.342,P=0.000)、淋巴结转移(r=0.197,P=0.018)和间质浸润深度(r=0.203,P=0.015)呈正相关。PCNA表达与临床分期(r=0.228,P=0.006)、病理分级(r=0.330,P=0.000)呈正相关。MMP-9和PCNA在宫颈癌组织中的表达强度呈正相关(r=0.228,P=0.006),二者表达一致的比例高达87.41%(125/143)。结论MMP-9和PCNA在宫颈癌中的异常表达与肿瘤的分化、侵袭和发展密切相关,联合检测二者的表达对于进一步理解宫颈癌的生物学行为和判断预后有一定价值。     

钼,铁,硫化合物激活曝氧的棕色固氮菌固氮酶钼铁蛋白的研究

曝氧后,棕色固氮菌(Azotobacter vinelandii)固氮酶钼铁蛋白的催化活性和圆二色信号都显著降低,而吸收光谱则显著增加。与钼、铁、硫化合物和二硫苏糖醇组成的重组溶液保温后,曝氢蛋白的圆二色信号和吸收光谱几乎完全恢复至天然状态的同时,乙炔还原活性也得到了显著的恢复,表明重组溶液可使曝氧蛋白中的 P-cluster和其它活性部位都得到了不同程度的修复。