Symmetrical directional cloning: An efficient method to prepare hairpin RNA interference constructs

RNA interference (RNAi) is a loss-of-function approach by which double-stranded RNA (dsRNA) initiates degradation of homologous mRNAs in a sequence specific manner. The dsRNA molecules can be produced in vitro or in vivo, and can be introduced to cells in a number of ways. Here we report a more efficient method for the cloning of inverted repeat DNA fragments into expression vectors that can be transcribed into effective dsRNA molecules in vivo or in vitro. This method, named Symmetrical Directional Cloning (SDC), takes the advantage of compatible non-palindromic restriction enezyme sites, which allow one to directionally clone a single PCR product in both the sense and antisense orientations together into a vector. SDC allows for the directional cloning of inverted repeats using a single PCR product; it requires only one cut site on each side of the loop. Hence this method is more cost effective and less time-consuming. At least 21 commercially available restriction endonucleases can be used as cloning sites for the SDC method. The efficacy of dsRNA expression vectors prepared by SDC has been demonstrated by targeting a negative regulator of the signaling pathway mediating the response of cells to phytohormone, gibberellins (GA), in the aleurone cells.     

Adelfa： a futuristic biodiesel fuel for DI diesel engines： a comparative experimental investigation

Biodiesel fuel is found to be a promising alter- native for the petroleum diesel based on the results published by the researchers for a decade. Biodiesel fuel is renewable and non degradable fuel. Many countries use biodiesel fuel for automotives to meet the crisis due to the depletion of the petroleum fuel and to meet the stringent emission norms. Various researches have been carried out with different bio- diesel fuels with vegetable oil as the source and appreciable results were reported. Few biodiesel fuels which have been already tested are Jatropha, Pongamia, Mahua, neem, cotton seed, etc. In this experimental work, Adelfa biodiesel blend is used as the test fuel. The emission and performance charac- teristics were compared with three other different biodiesel fuel blends. Appreciable results imply that Adelfa biodiesel （Nerium oil methyl ester） can be a futuristic biodiesel fuel, which has a good compatibility with the direct injection （DI） diesel engine without any major modification. Moreover, Adelfa can be cultivated in a non agricultural land with fewer sources of water. It is widely spread over all major countries of Asia. Experimental investigations have been carried out on a single cylinder DI diesel engine with standard engine speci- fications. In this experimental work, various Adelfa biodiesel blends is compared with reference fuel （diesel） to choose the best blend which gives a closer performance to diesel. The comparative analysis with other biodiesel fuels has also been done and results have been discussed.     

Temporal precision in the neural code and the timescales of natural vision

The timing of action potentials relative to sensory stimuli can be precise down to milliseconds in the visual system, even though the relevant timescales of natural vision are much slower. The existence of such precision contributes to a fundamental debate over the basis of the neural code and, specifically, what timescales are important for neural computation. Using recordings in the lateral geniculate nucleus, here we demonstrate that the relevant timescale of neuronal spike trains depends on the frequency content of the visual stimulus, and that 'relative', not absolute, precision is maintained both during spatially uniform white-noise visual stimuli and naturalistic movies. Using information-theoretic techniques, we demonstrate a clear role of relative precision, and show that the experimentally observed temporal structure in the neuronal response is necessary to represent accurately the more slowly changing visual world. By establishing a functional role of precision, we link visual neuron function on slow timescales to temporal structure in the response at faster timescales, and uncover a straightforward purpose of fine-timescale features of neuronal spike trains.     

Direct splitting of water under visible light irradiation with an oxide semiconductor photocatalyst.

The photocatalytic splitting of water into hydrogen and oxygen using solar energy is a potentially clean and renewable source for hydrogen fuel. The first photocatalysts suitable for water splitting, or for activating hydrogen production from carbohydrate compounds made by plants from water and carbon dioxide, were developed several decades ago. But these catalysts operate with ultraviolet light, which accounts for only 4% of the incoming solar energy and thus renders the overall process impractical. For this reason, considerable efforts have been invested in developing photocatalysts capable of using the less energetic but more abundant visible light, which accounts for about 43% of the incoming solar energy. However, systems that are sufficiently stable and efficient for practical use have not yet been realized. Here we show that doping of indium-tantalum-oxide with nickel yields a series of photocatalysts, In(1-x)Ni(x)TaO(4) (x = 0-0.2), which induces direct splitting of water into stoichiometric amounts of oxygen and hydrogen under visible light irradiation with a quantum yield of about 0.66%. Our findings suggest that the use of solar energy for photocatalytic water splitting might provide a viable source for 'clean' hydrogen fuel, once the catalytic efficiency of the semiconductor system has been improved by increasing its surface area and suitable modifications of the surface sites.     

A role for mitochondria in NLRP3 inflammasome activation

An inflammatory response initiated by the NLRP3 inflammasome is triggered by a variety of situations of host 'danger', including infection and metabolic dysregulation. Previous studies suggested that NLRP3 inflammasome activity is negatively regulated by autophagy and positively regulated by reactive oxygen species (ROS) derived from an uncharacterized organelle. Here we show that mitophagy/autophagy blockade leads to the accumulation of damaged, ROS-generating mitochondria, and this in turn activates the NLRP3 inflammasome. Resting NLRP3 localizes to endoplasmic reticulum structures, whereas on inflammasome activation both NLRP3 and its adaptor ASC redistribute to the perinuclear space where they co-localize with endoplasmic reticulum and mitochondria organelle clusters. Notably, both ROS generation and inflammasome activation are suppressed when mitochondrial activity is dysregulated by inhibition of the voltage-dependent anion channel. This indicates that NLRP3 inflammasome senses mitochondrial dysfunction and may explain the frequent association of mitochondrial damage with inflammatory diseases.     

断层相互作用产生同震库仑应力改变分布图像

讨论了同震库仑应力改变的基本含义,以及基于静应力位错理论求解同震库仑应力改变的边界元方法,对与发震断层相互平行和相互垂直两种典型构造模式下,断层相互作用产生的同震库仑应力改变进行了数值模拟研究,分别获取了相应条件下的同震库仑应力改变空间分布图像.结果表明:不同构造模式的断层相互作用,其产生的同震库仑应力改变分布图像存在较大差异,在分析和评价断层相互作用产生的同震库仑应力改变,及其对活动断层地震危险性的影响时,应该充分考虑待评价断层与发震断层之间的空间几何关系、断层的性质和产状等特定的构造条件.     

空间n边形中的正弦定理

利用向量代数中的一个基本等式进行推证,得出了在空间 ｎ 边形中正弦定理的表现形式。使文［１］,［２］,［３］更加完整     

Anthropogenic ocean acidification over the twenty-first century and its impact on calcifying organisms

Today's surface ocean is saturated with respect to calcium carbonate, but increasing atmospheric carbon dioxide concentrations are reducing ocean pH and carbonate ion concentrations, and thus the level of calcium carbonate saturation. Experimental evidence suggests that if these trends continue, key marine organisms--such as corals and some plankton--will have difficulty maintaining their external calcium carbonate skeletons. Here we use 13 models of the ocean-carbon cycle to assess calcium carbonate saturation under the IS92a 'business-as-usual' scenario for future emissions of anthropogenic carbon dioxide. In our projections, Southern Ocean surface waters will begin to become undersaturated with respect to aragonite, a metastable form of calcium carbonate, by the year 2050. By 2100, this undersaturation could extend throughout the entire Southern Ocean and into the subarctic Pacific Ocean. When live pteropods were exposed to our predicted level of undersaturation during a two-day shipboard experiment, their aragonite shells showed notable dissolution. Our findings indicate that conditions detrimental to high-latitude ecosystems could develop within decades, not centuries as suggested previously.     

Human gamma-chain genes are rearranged in leukaemic T cells and map to the short arm of chromosome 7

Three gene families that rearrange during the somatic development of T cells have been identified in the murine genome. Two of these gene families (alpha and beta) encode subunits of the antigen-specific T-cell receptor and are also present in the human genome. The third gene family, designated here as the gamma-chain gene family, is rearranged in murine cytolytic T cells but not in most helper T cells. Here we present evidence that the human genome also contains gamma-chain genes that undergo somatic rearrangement in leukaemia-derived T cells. Murine gamma-chain genes appear to be encoded in gene segments that are analogous to the immunoglobulin gene variable, constant and joining segments. There are two closely related constant-region gene segments in the human genome. One of the constant-region genes is deleted in all three T-cell leukaemias that we have studied. The two constant-region gamma-chain genes reside on the short arm of chromosome 7 (7p15); this region is involved in chromosomal rearrangements identified in T cells from individuals with the immunodeficiency syndrome ataxia telangiectasia and observed only rarely in routine cytogenetic analyses of normal individuals. This region is also a secondary site of beta-chain gene hybridization.