Regulation of the proliferative potential of cord blood long-term culture-initiating cells (LTC-IC) by different stromal cell lines: implications for LTC-IC measurement

BACKGROUND AND OBJECTIVE: Long-term culture-initiating cells (LTC-IC) are the best available approximation to an in vitro assay of stem cells in humans although they still represent a heterogeneous population in terms of proliferative capacity and sensitivity to different growth factors. Human umbilical cord blood (CB) is rich in hemopoietic progenitor cells, as measured by clonogenic assays and contains stem cells capable of reconstituting the marrow after ablation in clinical transplantation. We evaluated the influence of culture conditions on the in vitro behavior of LTC-IC from CB. DESIGN AND METHODS: LTC-IC were evaluated in long-term cultures, comparing two types of murine stromal cell lines: M2-10B4 and M2-10B4 transfected with cDNAs for human G-CSF and IL-3. RESULTS: Two and five fold higher numbers of terminally differentiated cells were produced during nine weeks of culture of CB mononuclear or CD34+ cells respectively, in cultures containing a M2-10B4 IL-3 G-CSF cell line compared to cultures containing the parental cell line. Likewise, a higher number of colony-forming cells (CFC) were detected in the supernatant of cultures with the transfected cell line. In contrast, the number of CFC generated within the stromal layer, after 5 or 9 weeks of culture, was significantly higher in cultures on M2-10B4 cells than those on M2-10B4 IL-3 G-CSF. INTERPRETATION AND CONCLUSIONS: Our results show that the proliferative capacity of CB LTC-IC can be strongly influenced by culture conditions and that the frequency of LTC-IC estimated using these cell lines as stromal support is not identical.     

Simulated and measured X‐band radar reflectivity of land in mountainous terrain using a fan‐beam antenna

A computer program for meteorological radar siting, previously developed for pencil‐beam antenna, long‐range, C‐band radars, has been adapted for fan‐beam antenna, short‐range, X‐band radars. The simulator uses topographic information in the form of a raster digital elevation model and a surface backscattering cross‐section per unit area at grazing angles derived from the literature. It is independent of specific radar sites and radar systems. Its use will enhance optimized scanning strategies, antenna design tailored to mountainous terrain and knowledge of land‐clutter measurements at low grazing angles. The raster‐based approach used in implementing the program makes it compatible with any desired spatial resolution. Simplicity and short simulation times have been pursued as primary goals during the planning and implementation phases of the routines. A preliminary validation of the simulator using backscattering data acquired in the Alps is presented in this letter.     

Scale-space and edge detection using anisotropic diffusion

A new definition of scale-space is suggested, and a class of algorithms used to realize a diffusion process is introduced. The diffusion coefficient is chosen to vary spatially in such a way as to encourage intraregion smoothing rather than interregion smoothing. It is shown that the `no new maxima should be generated at coarse scales' property of conventional scale space is preserved. As the region boundaries in the approach remain sharp, a high-quality edge detector which successfully exploits global information is obtained. Experimental results are shown on a number of images. Parallel hardware implementations are made feasible because the algorithm involves elementary, local operations replicated over the image     

Measuring and Predicting Object Importance

How important is a particular object in a photograph of a complex scene? We propose a definition of importance and present two methods for measuring object importance from human observers. Using this ground truth, we fit a function for predicting the importance of each object directly from a segmented image; our function combines a large number of object-related and image-related features. We validate our importance predictions on 2,841 objects and find that the most important objects may be identified automatically. We find that object position and size are particularly informative, while a popular measure of saliency is not.     

Rho-regulated signals induce apoptosis in vitro and in vivo by a p53-independent, but Bcl2 dependent pathway

Rho proteins are a branch of GTPases that belongs to the Ras superfamily which are critical elements of signal transduction pathways leading to a variety of cellular responses. This family of small GTPases has been involved in diverse biological functions such as cytoskeleton organization, cell growth and transformation, cell motility, migration, metastasis, and responses to stress. We report that several human Rho proteins including Rho A, Rho C and Rac 1, are capable of inducing apoptosis in different cell systems like murine NIH3T3 fibroblasts and the human erythroleukemia K562 cell line. Since K562 cells are devoid of p53, apoptosis induced by Rho in this system is independent of p53. Rho-dependent apoptosis is mediated by the generation of ceramides, and it is drastically inhibited by ectopic expression of Bcl2, both under in vitro and in vivo conditions. Furthermore, the human oncogenes vav and ost that have been shown to function as guanine exchange factors for Rho proteins, were also able to induce apoptosis under similar conditions. Finally, we also report that the levels of endogenous Rho proteins are increased when U937 myeloid leukemia cells are exposed to apoptosis-inducing conditions such as TNF alpha treatment. Furthermore, TNF alpha-induced apoptosis in these cells is inhibited by expression of a dominant negative mutant of Rac 1 but it is not affected by a similar mutant of Rho A. These results suggest that Rho proteins play an important role in the physiological regulation of the apoptotic response to stress-inducing agents.     

Metal ion-mediated substrate-assisted catalysis in type II restriction endonucleases

The 2.15-A resolution cocrystal structure of EcoRV endonuclease mutant T93A complexed with DNA and Ca2+ ions reveals two divalent metals bound in one of the active sites. One of these metals is ligated through an inner-sphere water molecule to the phosphate group located 3' to the scissile phosphate. A second inner-sphere water on this metal is positioned approximately in-line for attack on the scissile phosphate. This structure corroborates the observation that the pro-SP phosphoryl oxygen on the adjacent 3' phosphate cannot be modified without severe loss of catalytic efficiency. The structural equivalence of key groups, conserved in the active sites of EcoRV, EcoRI, PvuII, and BamHI endonucleases, suggests that ligation of a catalytic divalent metal ion to this phosphate may occur in many type II restriction enzymes. Together with previous cocrystal structures, these data allow construction of a detailed model for the pretransition state configuration in EcoRV. This model features three divalent metal ions per active site and invokes assistance in the bond-making step by a conserved lysine, which stabilizes the attacking hydroxide ion nucleophile.     

Challenges in supply chain redesign for the Circular Economy: a literature review and a multiple case study

Despite companies face several challenges when redesigning their supply chain for the Circular Economy, the literature lacks a systematisation of such challenges and of the ways to overcome them. Through a systematic literature review, this paper identifies and systematises 24 challenges that may hamper a supply chain redesign for the Circular Economy. Sixteen among these challenges are well known from research in related topics. On the contrary, the remaining eight are relatively new or take a different relevance within the Circular Economy context. A multiple case study in the household appliance supply chain is carried out, to explore how these challenges appear in practice and how companies may tackle them. The cases analysed involve actors at different supply chain levels, and findings suggest that a great degree of vertical integration by one actor in the supply chain is not a necessary condition for Circular Economy implementation. The empirical study, in conjunction with the literature analysis, leads to the development of a framework linking the challenges to specific levers that companies may pursue to overcome them. The framework can be seen as a reference for managers undertaking the path towards Circular Economy.     

Visual identification by signature tracking

We propose a new camera-based biometric: visual signature identification. We discuss the importance of the parameterization of the signatures in order to achieve good classification results, independently of variations in the position of the camera with respect to the writing surface. We show that affine arc-length parameterization performs better than conventional time and Euclidean arc-length ones. We find that the system verification performance is better than 4 percent error on skilled forgeries and 1 percent error on random forgeries, and that its recognition performance is better than 1 percent error rate, comparable to the best camera-based biometrics.     

Recursive 3-D Visual Motion Estimation Using Subspace Constraints

A structure from motion algorithm is described which recovers structure and camera position, modulo a projective ambiguity. Camera calibration is not required, and camera parameters such as focal length can be altered freely during motion. The structure is updated sequentially over an image sequence, in contrast to schemes which employ a batch process. A specialisation of the algorithm to recover structure and camera position modulo an affine transformation is described, together with a method to periodically update the affine coordinate frame to prevent drift over time. We describe the constraint used to obtain this specialisation.Structure is recovered from image corners detected and matched automatically and reliably in real image sequences. Results are shown for reference objects and indoor environments, and accuracy of recovered structure is fully evaluated and compared for a number of reconstruction schemes. A specific application of the work is demonstrated—affine structure is used to compute free space maps enabling navigation through unstructured environments and avoidance of obstacles. The path planning involves only affine constructions.