In vivo genome editing restores haemostasis in a mouse model of haemophilia

Editing of the human genome to correct disease-causing mutations is a promising approach for the treatment of genetic disorders. Genome editing improves on simple gene-replacement strategies by effecting in situ correction of a mutant gene, thus restoring normal gene function under the control of endogenous regulatory elements and reducing risks associated with random insertion into the genome. Gene-specific targeting has historically been limited to mouse embryonic stem cells. The development of zinc finger nucleases (ZFNs) has permitted efficient genome editing in transformed and primary cells that were previously thought to be intractable to such genetic manipulation. In vitro, ZFNs have been shown to promote efficient genome editing via homology-directed repair by inducing a site-specific double-strand break (DSB) at a target locus, but it is unclear whether ZFNs can induce DSBs and stimulate genome editing at a clinically meaningful level in vivo. Here we show that ZFNs are able to induce DSBs efficiently when delivered directly to mouse liver and that, when co-delivered with an appropriately designed gene-targeting vector, they can stimulate gene replacement through both homology-directed and homology-independent targeted gene insertion at the ZFN-specified locus. The level of gene targeting achieved was sufficient to correct the prolonged clotting times in a mouse model of haemophilia B, and remained persistent after induced liver regeneration. Thus, ZFN-driven gene correction can be achieved in vivo, raising the possibility of genome editing as a viable strategy for the treatment of genetic disease.     

Highly coupled ATP synthesis by F1-ATPase single molecules

F1-ATPase is the smallest known rotary motor, and it rotates in an anticlockwise direction as it hydrolyses ATP. Single-molecule experiments point towards three catalytic events per turn, in agreement with the molecular structure of the complex. The physiological function of F1 is ATP synthesis. In the ubiquitous F0F1 complex, this energetically uphill reaction is driven by F0, the partner motor of F1, which forces the backward (clockwise) rotation of F1, leading to ATP synthesis. Here, we have devised an experiment combining single-molecule manipulation and microfabrication techniques to measure the yield of this mechanochemical transformation. Single F1 molecules were enclosed in femtolitre-sized hermetic chambers and rotated in a clockwise direction using magnetic tweezers. When the magnetic field was switched off, the F1 molecule underwent anticlockwise rotation at a speed proportional to the amount of synthesized ATP. At 10 Hz, the mechanochemical coupling efficiency was low for the alpha3beta3gamma subcomplex (F1-epsilon)), but reached up to 77% after reconstitution with the epsilon-subunit (F1+epsilon)). We provide here direct evidence that F1 is designed to tightly couple its catalytic reactions with the mechanical rotation. Our results suggest that the epsilon-subunit has an essential function during ATP synthesis.     

Thermal radiation scanning tunnelling microscopy

In standard near-field scanning optical microscopy (NSOM), a subwavelength probe acts as an optical 'stethoscope' to map the near field produced at the sample surface by external illumination. This technique has been applied using visible, infrared, terahertz and gigahertz radiation to illuminate the sample, providing a resolution well beyond the diffraction limit. NSOM is well suited to study surface waves such as surface plasmons or surface-phonon polaritons. Using an aperture NSOM with visible laser illumination, a near-field interference pattern around a corral structure has been observed, whose features were similar to the scanning tunnelling microscope image of the electronic waves in a quantum corral. Here we describe an infrared NSOM that operates without any external illumination: it is a near-field analogue of a night-vision camera, making use of the thermal infrared evanescent fields emitted by the surface, and behaves as an optical scanning tunnelling microscope. We therefore term this instrument a 'thermal radiation scanning tunnelling microscope' (TRSTM). We show the first TRSTM images of thermally excited surface plasmons, and demonstrate spatial coherence effects in near-field thermal emission.     

Microstructural evolution of a recycled aluminum alloy deformed by equal channel angular pressing process

The microstructural evolution of a recycled aluminum alloy after equal channel angular pressing (ECAP) up to four passes was investigated using X-ray diffraction (XRD) analysis and transmission electron microscopy (TEM). Microhardness tests were performed to determine the associated changes in mechanical properties. An ultrafine-grained material has been obtained with a microstructure showing a mixture of highly strained crystallites. A high density of dislocations was achieved as a result of severe plastic deformation (SPD) through the die. Changes in mechanical behavior are also revealed after ECAP due to strain hardening. Thermal analysis and TEM micrographs obtained after annealing indicate the succession of the recovery, recrystallization, and grain growth phenomena. Moreover, the energy stored during ECAP may be related to the dislocation density introduced by SPD. We finally emphasize the role played by the precipitates in this alloy.     

A 'snowball Earth' climate triggered by continental break-up through changes in runoff

Geological and palaeomagnetic studies indicate that ice sheets may have reached the Equator at the end of the Proterozoic eon, 800 to 550 million years ago, leading to the suggestion of a fully ice-covered 'snowball Earth'. Climate model simulations indicate that such a snowball state for the Earth depends on anomalously low atmospheric carbon dioxide concentrations, in addition to the Sun being 6 per cent fainter than it is today. However, the mechanisms producing such low carbon dioxide concentrations remain controversial. Here we assess the effect of the palaeogeographic changes preceding the Sturtian glacial period, 750 million years ago, on the long-term evolution of atmospheric carbon dioxide levels using the coupled climate-geochemical model GEOCLIM. In our simulation, the continental break-up of Rodinia leads to an increase in runoff and hence consumption of carbon dioxide through continental weathering that decreases atmospheric carbon dioxide concentrations by 1,320 p.p.m. This indicates that tectonic changes could have triggered a progressive transition from a 'greenhouse' to an 'icehouse' climate during the Neoproterozoic era. When we combine these results with the concomitant weathering effect of the voluminous basaltic traps erupted throughout the break-up of Rodinia, our simulation results in a snowball glaciation.     

熔丝成型薄板表面粗糙度理论模型与实验验证

针对熔丝成型(fused filament fabrication， FFF)产品的表面质量缺陷问题，提出了一种FFF产品表面粗糙度的理论建模方法.以材料丝表面轮廓为研究对象，考虑样件表面的实际情况，分别建立了FFF产品垂直和平行于纤维方向的表面粗糙度理论模型;与此同时，利用激光显微镜完成了相关样件表面粗糙度的实验研究;通过对比理论分析与实验结果，验证了所建模型的正确性;理论模型的敏感性分析研究表明，增大挤出宽度或打印层厚度会增大FFF产品的表面粗糙度，而增大重叠区域宽度则会降低表面粗糙度.     

FDM薄板振动特性的理论与实验研究

熔融沉积成型(FDM)是一种能够直接打印出具有复杂几何形状零件的快速成型技术.然而,FDM在振动性能方面很难与传统加工方式相媲美,需要对其进行更为实际的振动特性分析,为此首次提出FDM 3D打印薄板的振动特性理论建模方法.以悬臂边界条件下FDM薄板为研究对象,基于经典层合板理论,采用双向梁函数组合法表示振型函数,并通过Ritz法求解获得了复合薄板的固有特性;搭建了FDM 3D打印薄板的固有特性测试实验台,以获得薄板的模态振型和固有频率;计算与实验结果验证了理论计算方法的正确性和可靠性.     

Prevention or acceleration of type 1 diabetes by viruses

Type 1 diabetes is an autoimmune disease characterized by the destruction of insulin-producing pancreatic β-cells. Even though extensive scientific research has yielded important insights into the immune mechanisms involved in pancreatic β-cell destruction, little is known about the events that trigger the autoimmune process. Recent epidemiological and experimental data suggest that environmental factors are involved in this process. In this review, we discuss the role of viruses as an environmental factor on the development of type 1 diabetes, and the immune mechanisms by which they can trigger or protect against this pathology.