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PubMed2006

Microarray-based identification of antigenic variants of foot-and-mouth disease virus: a bioinformatics quality assessment.

BACKGROUND: The evolution of viral quasispecies can influence viral pathogenesis and the response to antiviral treatments. Mutant clouds in infected organisms represent the first stage in the genetic and antigenic diversification of RNA viruses, such as foot and mouth disease virus (FMDV), an important animal pathogen. Antigenic variants of FMDV have been classically diagnosed by immunological or RT-PCR-based methods. DNA microarrays are becoming increasingly useful for the analysis of gene expression and single nucleotide polymorphisms (SNPs). Recently, a FMDV microarray was described to detect simultaneously the seven FMDV serotypes. These results encourage the development of new oligonucleotide microarrays to probe the fine genetic and antigenic composition of FMDV for diagnosis, vaccine design, and to gain insight into the molecular epidemiology of this pathogen. RESULTS: A FMDV microarray was designed and optimized to detect SNPs at a major antigenic site of the virus. A screening of point mutants of the genomic region encoding antigenic site A of FMDV C-S8c1 was achieved. The hybridization pattern of a mutant includes specific positive and negative signals as well as crosshybridization signals, which are of different intensity depending on the thermodynamic stability of each probe-target pair. Moreover, an array bioinformatic classification method was developed to evaluate the hybridization signals. This statistical analysis shows that the procedure allows a very accurate classification per variant genome. CONCLUSION: A specific approach based on a microarray platform aimed at distinguishing point mutants within an important determinant of antigenicity and host cell tropism, namely the G-H loop of capsid protein VP1, was developed. The procedure is of general applicability as a test for specificity and discriminatory power of microarray-based diagnostic procedures using multiple oligonucleotide probes.

Animals↗

PubMed2005

Effects of point configuration on the accuracy in 3D reconstruction from biplane images.

Two or more angiograms are being used frequently in medical imaging to reconstruct locations in three-dimensional (3D) space, e.g., for reconstruction of 3D vascular trees, implanted electrodes, or patient positioning. A number of techniques have been proposed for this task. In this simulation study, we investigate the effect of the shape of the configuration of the points in 3D (the "cloud" of points) on reconstruction errors for one of these techniques developed in our laboratory. Five types of configurations (a ball, an elongated ellipsoid (cigar), flattened ball (pancake), flattened cigar, and a flattened ball with a single distant point) are used in the evaluations. For each shape, 100 random configurations were generated, with point coordinates chosen from Gaussian distributions having a covariance matrix corresponding to the desired shape. The 3D data were projected into the image planes using a known imaging geometry. Gaussian distributed errors were introduced in the x and y coordinates of these projected points. Gaussian distributed errors were also introduced into the gantry information used to calculate the initial imaging geometry. The imaging geometries and 3D positions were iteratively refined using the enhanced-Metz-Fencil technique. The image data were also used to evaluate the feasible R-t solution volume. The 3D errors between the calculated and true positions were determined. The effects of the shape of the configuration, the number of points, the initial geometry error, and the input image error were evaluated. The results for the number of points, initial geometry error, and image error are in agreement with previously reported results, i.e., increasing the number of points and reducing initial geometry and/or image error, improves the accuracy of the reconstructed data. The shape of the 3D configuration of points also affects the error of reconstructed 3D configuration; specifically, errors decrease as the "volume" of the 3D configuration increases, as would be intuitively expected, and shapes with larger spread, such as spherical shapes, yield more accurate reconstructions. These results are in agreement with an analysis of the solution volume of feasible geometries and could be used to guide selection of points for reconstruction of 3D configurations from two views.

Coronary Angiography↗

PubMed1996

Synaptic structural complexity as a factor enhancing probability of calcium-mediated transmitter release.

1. In a model synaptic system, the excitatory neuromuscular junction of the freshwater crayfish, the nerve terminals possess synapses that vary in structural complexity, with numbers of active zones ranging from zero to five. Active zones on individual synapses show a wide range of separation distances. We tested the hypothesis that two active zones of a single synapse in close proximity can enhance the localized increase in free calcium ion concentration, thus enhancing the probability of neurotransmission at that synapse. We evaluated the increase in calcium ion concentration as a function of distance between adjacent active zones. 2. To test this hypothesis, a reaction-diffusion model for Ca2+ entering the presynaptic terminals was used. This test was used because 1) present measurement techniques are inadequate to resolve quantitatively the highly localized, transient calcium microdomains at synaptic active zones; and 2) there is presently no suitable preparation for physiological recording from isolated synapses with varying distances between active zones. Included in the model were intracellular buffer and a typical distribution of voltage-activated Ca2+ channels for an active zone, estimated from freeze-fracture micrographs. 3. The model indicated that localized Ca2+ clouds from discrete active zones can overlap to create spatial enhancement of Ca2+ concentration. The degree of interaction between two active zones depends on the distance between them. When two typical active zones are separated by < or = 200 nm, the maximum intracellular Ca2+ concentration ([Ca2+]i) is greater at 1) the midpoint between them, and 2) the center of each one, than at the corresponding positions for a single isolated active zone. Enhanced [Ca2+]i at the edge of the active zone where "docked" synaptic vesicles occur would be expected to have an effect on transmitter release. 4. When the model includes no intracellular buffer, the increase in [Ca2+]i is a linear function of calcium channel current, but is a nonlinear function of the number of conducting calcium channels in an active zone. With immobile buffer included, the increase in [Ca2+]i is nonlinear with respect to both channel current and number of conducting channels. 5. Inclusion of immobile buffer in the model provides "released" residual calcium that slowly accumulates during a train of current pulses. Released residual calcium accumulates more rapidly at paired active zones separated by < or = 200 nm that at single isolated active zones. 6. We propose that the probability of release is enhanced at synapses with closely associated active zones. Synapses of this type ("complex" synapses) could be selectively recruited when the neuron is active at low frequencies. At higher frequencies of neuronal activity, more distant active zones may interact and acquire a greater probability of releasing quanta. This would provide the nerve terminal with one component of a mechanism for frequency facilitation, because the number of quanta released by the terminal as a whole would increase with frequency. Thus variation in synaptic complexity in a nerve terminal provides a mechanism for short-term plasticity of transmitter release.

Animals↗

PubMed2002

Moyamoya disease in a Hispanic child: a case report.

BACKGROUND: Moyamoya disease was initially described by Suzuki and Takaku in 1963 as a radiographic phenomenon relating to the tiny collateral vessels characteristic of the disease that resemble a cloud or puff of smoke. The disease is rare and initially it was believed that the disease was confined to the Japanese population. It consists of occlusive vascular disease at the Circle of Willis with a tendency toward multiple ischemic neurological events and small strokes. In older populations it can often be associated with further vascular degeneration and intracerebral hemorrhage. This paper discusses the diagnosis, treatment, and management of moyamoya disease in a Hispanic child. CASE DESCRIPTION: The case of a Hispanic child who presented with transient ischemic attacks over a period of 1 year is reported. Magnetic resonance imaging (MRI) revealed occlusive vascular disease in the posterior Circle of Willis. Digital subtraction cerebral angiography showed vascular occlusion at the base of the skull with collateral leptomeningeal and posterior circulation contribution in a pattern typical of moyamoya disease. Technetium was injected for a SPECT study demonstrating less uptake in the left frontal and left parieto-occipital regions. The patient underwent a left superficial temporal-to-middle cerebral artery anastamosis followed by a right-sided anastamosis in a second operation. The patient tolerated the cerebral revascularization and was symptom-free at 6-month follow-up. Cerebral angiography demonstrated improved perfusion in both cerebral hemispheres postoperatively. CONCLUSION: This article reports the occurrence of moyamoya in a Hispanic child. It illustrates the improved perfusion postoperatively as seen on digital subtraction cerebral angiography. Direct revascularization is felt to be difficult in children and alternatives such as encephaloduroarteriosynangiosis have been advocated. Direct revascularization was effective in treating moyamoya disease in this instance. Most of the discussion of moyamoya disease has been focused on the Japanese and Far East population. This report confirms the entity as occurring in a Hispanic individual in the United States with no known Japanese ancestry.

Brain↗