Due to the lack of an adequate conventional therapy against lower limb ischemia, gene transfer for therapeutic angiogenesis is seen as an attractive alternative. However, the possibility of side effects, due to the expression of large amounts of angiogenic factors, justifies the design of devices that express synergistic molecules in low controlled doses. We have developed an internal ribosome entry site (IRES)–based bicistronic vector expressing two angiogenic molecules, fibroblast growth factor 2 (FGF2), and Cyr61. Through electrotransfer into the ApoE−/− mice hindlimb ischemic muscle model, we show that the IRES-based vector gives more stable expression than either monocistronic plasmid. Furthermore, laser Doppler analysis, arteriography, and immunochemistry clearly show that the bicistronic vector promotes a more abundant and functional revascularization than the monocistronic vectors, despite the fact that the bicistronic system produces 5–10 times less of each angiogenic molecule. Furthermore, although the monocistronic Cyr61 vector accelerates B16 melanoma growth in mice, the bicistronic vector is devoid of such side effects. Our results show an active cooperation of FGF2 and Cyr61 in therapeutic angiogenesis of hindlimb ischemia, and validate the use of IRES-based bicistronic vectors for the coexpression of controlled low doses of therapeutic molecules, providing perspectives for a safer gene therapy of lower limb ischemia.
Abstract. The Mars Pathfinder mission used a unique capability to rapidly generate and interactively display three-dimensional (3-D) photorealistic virtual reality (VR) models of the Martian surface. An interactive terrain visualization system creates and renders digital terrain models produced from stereo images taken by the Imager for Mars Pathfinder (IMP) camera. The stereo pipeline, an automated machine vision algorithm, correlates features between the left and right images to determine their disparity and computes the corresponding positions using the known camera geometry. These positions are connected to form a polygonal mesh upon which IMP images are overlaid as textures. During the Pathfinder mission, VR models were produced and displayed almost as fast as images were received. The VR models were viewed using MarsMap, an interface that allows the model to be viewed from any perspective driven by a standard three-button computer mouse. MarsMap incorporates graphical representations of the lander and rover and the sequence and spatial locations at which rover data were taken. Graphical models of the rover were placed in the model to indicate the rover position at the end of each day of the mission. Images taken by Sojourner cameras are projected into the model as 2-D "billboards" to show their proper perspective. Distance and angle measurements can be made on features viewed in the model using a mouse-driven 3-D cursor and a point-andclick interface. MarsMap was used to assist with archiving and planning Sojourner activities and to make detailed measurements of surface features such as wind streaks and rock size and orientation that are difficult to perform using 2-D images. Superresolution image processing is a computational method for improving image resolution by a factor of n 1/2 by combining n independent images. This technique was used on Pathfinder to obtain better resolved images of Martian surface features. We show results from superresolving IMP camera images of six targets including near-and far-field objects and discuss how the resolution improvement aids interpretation. Similar flood deposits can be seen on both of the Twin Peaks that cannot be resolved in raw images. Millimeter-sized pits are resolved on the rocks Wedge and Halfdome. Other rocks at the Pathfinder site exhibit fine-scale layering that is otherwise invisible. Use of the method resulted in the probable discovery of an artifact of intelligent life on Mars: a part of the Pathfinder spacecraft. IntroductionPathfinder was the first rover mission to Mars, but it will not be the last. Rover missions are able to perform many of the functions of a field geologist [Stoker, 1996[Stoker, , 1998]. For example, a field geologist is able to look around, construct a mental three-dimensional (3-D) model of the nearby surroundings; perform measurements of nearby objects; determine slopes, strike, and dip of the distant terrain; and construct detailed maps of the region. The geologist also uses binoculars to get improved resolution of distant fe...
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