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Biomedical subjects

V Grau

Publications and source records attributed to V Grau.

At least 19 recordsLinked to original sources

Zero-temperature equation of state of two-dimensional 3He.

The equation of state of two-dimensional 3He at zero temperature has been calculated using the diffusion Monte Carlo method. By means of a combination of the fixed-node and released-node techniques, it is shown that backflow correlations provide a very accurate equation of state. The results prove unambiguously the non-self-bound character of two-dimensional 3He due to its Fermi statistics. We present solid evidence that the gas phase, predicted for the two-dimensional system, can be extrapolated to the case of 3He adsorbed on a strong substrate such as graphite.

Journal Article↗

Accumulating monocytes in the vasculature of rat renal allografts: phenotype, cytokine, inducible no synthase, and tissue factor mRNA expression.

BACKGROUND: Necrotic patches and hemorrhagic lesions develop in the renal tissue between day 4 and day 5 after transplantation of fully allogeneic DA rat kidneys to LEW recipients. These lesions are at least in part due to destruction and obstruction of blood vessels. Damage of graft endothelial cells and blood coagulation are likely to be mediated by intravascular graft leukocytes. However, this cell population has not been thoroughly characterized before. METHODS: We perfused untreated control kidneys, renal isografts, and allografts on day 4 after transplantation with phosphate-buffered saline/ethylenediaminetetraacetic acid to harvest leukocytes from both the blood stream as well as from the marginal intravascular pool. The mRNA expression of typical products of activated monocytes was analyzed in reverse-transcriptase polymerase chain reaction experiments. Graft monocytes were purified and their immunophenotype was investigated by flow cytometry. RESULTS: Allograft rejection led to a 10-fold increase in the number of intravascular graft leukocytes compared to isografts. A mean number of about 100x10(6) leukocytes was harvested from a single allogeneic kidney, about 73% of these cells were monocytes and most of them displayed an activated phenotype. Compared to isografts, intravascular allograft leukocytes displayed an increased expression of tumor necrosis factor-alpha, inducible NO synthase and tissue factor. CONCLUSIONS: Our study shows that large numbers of activated monocytes accumulate inside allograft vessels. As they express genes the products of which might damage the allograft by inducing cell death or thrombosis, we speculate that they directly participate in allograft destruction.

Animals↗

Automatic localization of cephalometric Landmarks.

A system for automatic detection of cephalometric landmarks is presented. Landmark detection is carried out in two steps: a line detection module searches for significant, well-contrasted lines of the image, such as the jaw line or the nasal spine. The landmark detection module uses the lines located in the first module to determine the search areas and then applies a pattern detection algorithm, based on mathematical morphology techniques. Relations between landmarks and lines are determined by means of a training process. The system has been tested for the detection of 17 landmarks on 20 images: more than 90% of the landmarks are accurately identified.

Algorithms↗

Monocytes in the rat: phenotype and function during acute allograft rejection.

Cells of the monocyte/macrophage system originate from the bone marrow, reach the organs via the blood, immigrate through postcapillary venules and further differentiate into organ-specific tissue macrophages. In rats and other species, activated monocytes/macrophages aggravate autoimmune reactions, rejection of non-vascularized allografts and chronic allograft rejection. It is very likely that they also contribute to acute allograft destruction. So far it has been impossible to distinguish the function of monocytes from that of macrophages, because cell phenotypes and their alterations upon activation are ill-defined. We have thus begun to characterize the ex vivo phenotype and function of rat monocytes in the normal state and during renal allograft rejection. Monocytes are recovered from both the central and the marginal blood pool by perfusing either the recipient's circulation or the allograft vasculature. Rat monocytes have a unique surface phenotype. During allograft rejection or after infusion of interferon-gamma they up-regulate class II MHC molecules, CD161 (NKR-P1A), CD62L and CD8, while CD4 and CD43 are down-modulated. Activated perfusate monocytes exert increased in vitro cytotoxicity against tumour targets, which differs from that of NK cells. We speculate that activated monocytes contribute to kidney allograft destruction by directly damaging endothelial cells or by promoting intravascular coagulation.

Animals↗

Monocytes in the rat.

We review our methods for definition and phenotypical characterisation of normal and activated rat monocytes. To obtain a comprehensive sample of all blood monocytes including cells from the marginal pool of the blood stream, we extensively perfuse the extrapulmonary circulation with cold PBS/EDTA. Normal rat monocytes are isolated from untreated specified pathogen-free male LEW rats. In vivo activated monocytes are investigated after three days of infusion of recombinant IFN-gamma or during acute renal allograft rejection. Rat monocytes are defined by reactivity with mAbs ED1 and ED9, detecting a lysosomal membrane antigen and a member of the signal-regulatory protein family, respectively, as well as by expression of CD11b. Concomitantly rat monocytes are characterized by the absence of CD5, the absence of the B cell form of CD45R, and the absence of reactivity with mAb RP-1. The majority of the monocytes from untreated LEW rats are CD4+, CD11a(high), CD18high, CD43high, CD62-L-, CD161-, and MHC class II-. Upon stimulation of the immune system in vivo, a second monocyte population increases in number. These cells have a larger diameter and an increased granularity. They are CD4-, CD11a(int), CD18int, CD43low, CD62-L+, CD161int, and MHC class II+. Although some reagents are not yet available (e.g. antibodies against rat CD14 and CD16), rat monocytes can be defined and their state of activation can be characterized. The functionally important population of monocytes, which have already marginated, is accessible by perfusion and relatively high monocyte numbers are isolated per rat. As specified pathogen-free rats are available and numerous experimental systems involving acute or chronic inflammation have been established in rats, differentially activated monocytes may be investigated. The rat is thus a suitable experimental animal for basic research on monocytes.

Animals↗

Chemokine expression during acute rejection of rat kidneys.

During acute rejection of fully allogeneic rat renal allografts, few neutrophil granulocytes are detected, whereas an abundant infiltrate of macrophages and T lymphocytes becomes apparent. The mechanisms leading to this specific pattern of infiltration are not understood. We performed a sequential daily Northern blot analysis of the mRNA expression of the CC-chemokines MCP-1, MIP-1alpha and RANTES and of the CXC-chemokines GRO/KC and MIP-2 in rat renal isografts (LEW --> LEW, n = 1 per day) and allografts during acute rejection (DA --> LEW, n = 3 per day). MCP-1 gene expression strongly increased on days 3-4 after allotransplantation and returned to control levels on day 6. The expression of MIP-1alpha and RANTES continuously rose until day 3-4 and remained stable thereafter. Isografts displayed minor changes in CC-chemokine expression. In contrast to CC-chemokines, GRO/KC was expressed in low amounts during rejection and MIP-2 mRNA remained undetectable. In conclusion, the expression of the CC-chemokines MCP-1, MIP-1 and RANTES was clearly upregulated during rejection, whereas the mRNA of the CXC-chemokines MIP-2 and GRO/KC was not detected at all or remained at low levels. This pattern of chemokine gene expression is in good accordance with the predominant mononuclear leukocyte infiltrate in allografts.

Animals↗

Multiresolution segmentation of three-dimensional medical images using mathematical morphology techniques.

A semi-automatic method for three-dimensional segmentation of medical images is proposed. A multiresolution representation is achieved through the application of morphological filters, which assures causality for image extrema. This allows for a compact scale space representation, in which each extremum is assigned a scale value. Interactive selection of the interesting extrema of the image is carried out, aided by this scale information and other relevant features. Extrema selected are then used as markers in three-dimensional watersheds calculation. The system has been developed and tested under low cost platforms, and can be the base for totally automatic, knowledge based segmentation systems.

Artificial Intelligence↗

A new efficient method for 3D registration using human brain atlases.

In this article a new efficient method in deformable brain atlases is proposed. To achieve this goal we use the composition of two applications, an affine transformation and other one. In this second application we have used three different approaches. One approach that has been used and two new approaches. We use a set of points to be identified both in brain atlas and in patient's brain for registration. The three approach present an advantage: the mathematical solution obtained makes it to be very fast, almost immediate. Several results have been obtained and they have been classified as very satisfactory by the neurosurgeons that have tested our system.

Brain Mapping↗

Interactive image-guided surgery system with high-performance computing capabilities on low-cost workstations: a prototype.

We present a new frameless stereotatic system prototype that has been initially validated in functional neurosurgery operations and that makes use of an optical position tracker for image-guided neurosurgery. Several devices for tracking different surgical instruments have been designed and manufactured. These devices include an array of infrared light-emitting diodes that are tracked by three charge-coupled device cameras. The system presents several new approaches for surgery planning. For high-quality 3D images of the patient's anatomy, we have developed a parallel version of a volume-rendering algorithm, thus enabling real-time 3D anatomy manipulation on low-cost PC workstations. In order to test the accuracy of the system, the localization of the target by means of a stereotatic frame has been compared with frameless techniques, obtaining a difference of about 1 +/- 1 mm.

Algorithms↗

A system for the simulation and planning of orthodontic treatment using a low cost 3D laser scanner for dental anatomy capturing.

The detection and correction of malocclusions and other dental abnormalities is a significant area of work in orthodontic diagnosis. To assess the quality of occlusion between the teeth the orthodontist has to estimate distances between specific points located on the teeth of both arches. Distance measuring is based on the observation, by the orthodontist, of a plaster model of the mouth. Gathering of information required to make the diagnosis is a time consuming and costly operation. On the other hand, obtaining and manipulation of plaster casts constitute a huge problem in clinics, due to both the large space needed and high costs associated with plaster casts manufacturing. For this problem we present a new system for three-dimensional orthodontic treatment planning and movement of teeth. We describe a computer vision technique for the acquisition and processing of three-dimensional images of the profile of hydrocolloids dental imprints taken by mean of a own developed 3D laser scanner. Profile measurement is based on the triangulation method which detects deformation of the projection of a laser line on the dental imprints. The system is computer-controlled and designed to achieve depth and lateral resolutions of 0.1 mm and 0.2 mm, respectively, within a depth range of 40 mm. The developed diagnosis software system (named MAGALLANES) and the 3D laser scanner (named 3DENT) are both commercially available and have been designed to replace manual measurement methods, which use costly plaster models, with computer measurements methods and teeth movement simulation using cheap hydrocolloid dental wafers. This procedure will reduce the cost and acquisition time of orthodontic data and facilitate the conduct of epidemiological studies.

Adult↗

Dynamics of monocytes/macrophages and T lymphocytes in acutely rejecting rat renal allografts.

We examined the infiltration of acutely rejecting renal allografts (DA-->LEW) by ED1+ and ED2+ macrophages and T lymphocytes at intervals of 24 h after transplantation. Donor and recipient macrophages were differentiated by MHC class II antigen expression in double-staining experiments with ED1. Proliferation was assayed after pulse-labelling with BrdU. We subdivided allograft infiltration into three consecutive phases: 1) During phase I on days 1 to 2 after allogeneic kidney transplantation, perivascular infiltrates developed that contained numerous donor and recipient macrophages. Allograft rejection could already be diagnosed 24 h after transplantation by perivascular infiltration of T lymphocytes, whereas T cells were rarely found in isografts. 2) Phase II of allograft rejection from day 3 to 4 was characterized by massive propagation of the infiltrate. About equal numbers of interstitial donor and recipient macrophages were counted. Both macrophages and T lymphocytes proliferated in situ and macrophages outnumbered T cells until complete rejection. 3) During phase III the allograft was destroyed. Large intravascular monocytes surprisingly expressed the ED2 antigen. In the interstitium of viable graft regions, the population of recipient macrophages grew, whereas the population of donor macrophages and of T lymphocytes decreased.

Acute Disease↗

De novo expression of MHC class II molecules by microglial cells during acute rat renal allograft rejection.

The expression of MHC class I and class II molecules in the cerebral cortex of rats was investigated at daily intervals from day 3 to day 6 after fully allogeneic (DA-->LEW) and isogeneic (LEW-->LEW) kidney transplantation. MHC class II molecules were temporarily induced on the previously negative microglial cells and on the endothelia of arterioles and venules during acute rejection. On the endothelia of all brain vessels MHC class I expression was enhanced. MHC class I+ cells with microglial cell morphology were discernible within the diffusely MHC class I+ brain parenchyma. In contrast, the brain parenchyma of isograft recipients and untreated control animals did not express detectable levels of MHC molecules. In conclusion, we demonstrate that a strong immune reaction in the periphery is able to activate microglial cells in the central nervous system.

Acute Disease↗

An advanced system for the simulation and planning of orthodontic treatment.

This paper presents a new system for three-dimensional (3-D) orthodontic treatment planning and movement of teeth. We describe a computer vision technique for the acquisition and processing of 3-D images of the profile of hydrocolloid dental imprints. Profile measurement is based on the triangulation method which detects deformation of the projection of a laser line on the dental imprints. The system is computer-controlled and designed to achieve depth and lateral resolutions of 0.1 and 0.2 mm, respectively, within a depth range of 40 mm. The 3-D image of the imprint is segmented in order to identify different teeth. Two operators are presented: one for the detection of molars and premolars based on a directional gradient, and one for incisors and canines based on 3-D registration with dental models contained in a database. We apply these 3-D dental models to simulate the 3-D movement of teeth, including rotations, during orthodontic treatment. With this objective, we have developed an original simplified model of arch-wire behaviour and a viscoplastic behaviour law for the alveolar bone in order to simulate teeth displacements during orthodontic treatment. The contribution of the paper is part of a diagnosis system (called MAGALLANES) that is designed to replace manual measurement methods, which use costly plaster models, with computer measurement methods and teeth movement simulation using cheap hydrocolloid dental wafers. This procedure will reduce the cost and acquisition time of orthodontic data and facilitate the conduct of epidemiological studies.

Biomechanical Phenomena↗

Phenotype of rat monocytes during acute kidney allograft rejection: increased expression of NKR-P1 and reduction of CD43.

Monocytes and macrophages mediate cytotoxicity after appropriate activation and thus represent effectors secondary to T lymphocytes and natural killer (NK) cells. However, very little is known about the role of activated monocytes in organ allograft rejection. In this study, isogeneic (LEW to LEW) and fully allogeneic (DA to LEW) rat kidneys were grafted to bilaterally nephrectomized recipients. Four days after transplantation a comprehensive sample of leucocytes was recovered by perfusion of the recipients' vasculature with phosphate-buffered saline containing EDTA (PBS/EDTA). Monocytes were enriched by density gradient centrifugation and their physical parameters and immunophenotype were investigated by flow cytometry in comparison with untreated, specified pathogen-free (SPF) LEW rats. Isotransplantation and allotransplantation of kidneys dramatically increased the absolute number of intravascular monocytes in the recipient. Analysis of NKR-P1 (CD161), CD4, CD62L, CD43, CD11a, CD18 and MHC class II expression identified at least two monocyte populations in all experimental groups. In graft recipients it was evident that activated monocytes were able to express and upregulate NKR-P1 and CD8, which have not been detected in these cells to date. If activated monocytes utilize NKR-P1 and CD8, by analogy to NK cells and lymphocytes these cells may be endowed with additional pathways to upregulate cytolytic functions and effect allograft damage.

Animals↗

Activation of microglial and endothelial cells in the rat brain after treatment with interferon-gamma in vivo.

Activation of microglial cells occurs during the pathogenesis of various neurologic diseases. However, the mechanisms of activation in vivo are still elusive. We infused adult rats intravenously with interferon-gamma and demonstrated microglial cell activation after three days of treatment. We show for the first time that microglial cells proliferate inside the brain in response to a circulating cytokine. Microglial cells were induced to express major histocompatibility (MHC) class I and class II antigens and small amounts of leukocyte function-associated molecule 1 (LFA-1, CD11a). On endothelial cells of the brain, MHC class I and class II antigens and intercellular adhesion molecule 1 (ICAM-1, CD54) were enhanced.

Animals↗

Rat monocytes up-regulate NKR-P1A and down-modulate CD4 and CD43 during activation in vivo: monocyte subpopulations in normal and IFN-gamma-treated rats.

LEW rats were treated intravenously with recombinant rat interferon-gamma (IFN-gamma) for 3 days to achieve intravascular accumulation, proliferation, and activation of monocytes. Monocytes, defined by their expression of the ED1, ED9, and Ox41 antigens, were recovered from the vasculature by perfusion with PBS/EDTA, subsequently depleted of erythrocytes and granulocytes by Percoll density gradient centrifugation, and analyzed by flow cytometry and immunocytology. In untreated and control-infused specified pathogen-free (SPF) rats, lymphocytes and monocytes formed overlapping cell populations with respect to size and internal granularity. At least two intravascular monocyte subsets, probably central and marginating cells, were distinguished by their size and differential expression of CD43, CD4, CD11a, CD18, and L-selectin. It is interesting to note that a fraction of the monocytes in normal and control-infused animals carried the NKR-P1A molecule. IFN-gamma treatment provoked a duplication of monocyte size and granularity. Both the number of positive monocytes and the level of expression of NKR-P1A strongly increased after IFN-gamma infusion, whereas CD43 (leukosialin) and CD4 were impressively down-regulated. NKR-P1A+ L-selectin+ CD43low CD4- monocytes also occur in the vasculature of rats during immune reactions in vivo. We speculate that these cells are involved in organ damage and that their number is controlled by activation-induced cell death within the vessels.

Animals↗