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An automated cell analysis sensing system based on a microfabricated rheoscope for the study of red blood cells physiology.

An automated rheoscope has been developed, utilizing a microfabricated glass flow cell, high speed camera and advanced image-processing software. RBCs suspended in a high viscosity medium were filmed flowing through a microchannel. Under these conditions, RBCs exhibit different orientations and deformations according to their location in the velocity profile. The rheoscope system produces valuable data such as velocity profile of RBCs, spatial distribution within a microchannel and deformation index (DI) curves. The variation of DI across the channel height, due to change in shear stress, was measured carrying implications for diffractometry methods. These curves of DI were taken at a constant flow rate and cover most of the relevant shear stress spectrum. This is an improvement of the existing techniques for deformability measurements and may serve as a diagnostic tool for certain blood disorders. The DI curves were compared to measurements of the flowing RBCs velocity profile. In addition, we found that RBCs flowing in a microchannel are mostly gathered in the center of the flow and maintain a characteristic spatial distribution. The spatial distribution in this region changes slightly with increasing flow rate. Hence, the system described, provides means for examining the behavior of individual RBCs, and may serve as a microfabricated diagnostic device for deformability measurement.

Biosensing Techniques↗

A new gel using super absorbent polymer for mapping the spatial dose distributions of electron beams by MR imager.

A technique for mapping the spatial dose distribution with a magnetic resonance imager is presented. A ferrous sulphate solution with sulfuric acid was used as the detecting medium for radiation dose. To make a gel of the solution for filling up a cubic phantom, we developed a new gel component that is combined with a super absorbent polymer (Sumikagel N-100) and a cross-linked dextran gel (Sephadex G-200). In order to make the application for radiation treatment planning, mapping of the dose distribution was carried out using a Unix computer.

Electrons↗

A new method to assess the fluidodynamic behaviour of an angiographic contrast agent.

PURPOSE: To propose a new method for the assessment of the fluidodynamic behaviour of angiographic contrast agents. The method enables measurement of the spatial distribution and time dependence of the contrast agent along a pseudo-vessel on images obtained with an X-ray image intensifier. MATERIAL AND METHODS: A particular phantom was devised consisting of a plexiglas box with an insert into which a latex tube with a 0.4 cm in diameter was placed in order to simulate the tortuous flow of a blood vessel. The box, which is filled with water to simulate the thickness of a normal patient, is connected to an injection and pumping system which serve to keep the contrast agent flowing in the pseudo-vessel tube. The pseudo-vessel tube was filled with plain water in one case and with saline solution in another case to assess their different dilution capabilities. The phantom and the flow of contrast agent were imaged with a conventional X-ray image intensifier system and the images were stored in digital format during 35 second acquisitions at a speed of 4 frames per second; for any frame it is possible to measure the mathematical contrast in any position in the image. Further, a diagram showing the time dependence of the spatial distribution of the mathematical contrast is proposed. The X axis shows the spatial distribution of the mathematical contrast, whereas the Y axis shows its temporal variation with a gray level proportional to the mathematical contrast value. By building an horizontal profile of this diagram one can obtain the spatial distribution at a fixed time, while by building a vertical profile one can obtain the temporal variation at a fixed point. Several different contrast agents were so tested. RESULTS: The proposed method allows different fluido-dynamic behaviour patterns of contrast agents and flowing media to be shown. Owing to the different chemical characteristics of water and saline solution these media have different dilution capabilities (higher for water) and this is well demonstrated by the diagram profiles obtained for each. The system has also allowed the detection of a particular behaviour of some contrast agents, whose spatial distribution was non uniform even in the last frames, thus showing a tendency to maintain their bolus-nature over time. An interesting feature which can be noticed in all the temporal profiles is the presence of a "pre-bolus", i.e. the contrast is higher at the very beginning of the flow, then decreases and after some time starts to increase again. Though the initial contrast value obviously depends on the iodine concentration employed, the method shows the contrast variation as a function of time is different for different contrast agents. CONCLUSIONS: The method and the equipment proposed provide a good description of the fluidodynamic behaviour of different contrast agents, but do not constitute a reference method for testing haemodynamic behaviour which, "in vivo", is obviously affected by several other chemical and metabolic factors. However, the method allows evaluation of the contrast agents from a physical and fluidodynamic point of view, showing that the iodine content is not the only feature affecting their behaviour. The method can be used in quality control to test the constancy of the physical behaviour of different contrast agents.

Angiography↗

Covariances among join-count spatial autocorrelation measures.

Spatial distributions of biological variables are often well-characterized with pairwise measures of spatial autocorrelation. In this article, the probability theory for products and covariances of join-count spatial autocorrelation measures are developed for spatial distributions of multiple nominal (e.g. species or genotypes) types. This more fully describes the joint distributions of pairwise measures in spatial distributions of multiple (i.e. more than two) types. An example is given on how the covariances can be used for finding standard errors of weighted averages of join-counts in spatial autocorrelation analysis of more than two types, as is typical for genetic data for multiallelic loci.

Analysis of Variance↗

[The distribution of the macrobenthos of the White Sea littoral in different spatial scales].

Spatial distribution of macrobenthos of middle intertidal zone was studied in scale from centimetres to 30 kilometres along the coastline. The community structure and distribution of the 5 most abundant species (Hydrobia ulvae, Mya arenaria, Macoma baltica. Peloscolex benedeni, Arenicola marina) were considered. Spatial heterogeneity of macrobenthos, estimated as mean dissimilarity between samples, kept constant in scale of centimetres--meters, but increased significantly when enlarged area is considered. Patterns of many species changed with scale from random mosaic to more or less pronounced patchiness, whereas the density of H. ulvae and the structure of the whole community demonstrated fractal (self-similar) patch pattern in wide range of scale from dozens of meters to several kilometres. Spatial correlations between species (the composition of assemblages) and between species and environmental factors were also scale dependent. Some possible effects of scale on the observed spatial distribution of benthos are discussed, and multiscaled analysis of biotic heterogeneity is concluded to be very fruitful.

Animals↗

Spatiotemporal visualization of long-term potentiation and depression in the hippocampal CA1 area.

Long-term potentiation (LTP) in the CA1 area of the hippocampus depends critically on the statistical characteristics of its stimulus. The ability of optical imaging to record spatial distribution has made it possible to examine systematically the effect of higher-order statistical characteristics, such as the correlation between successive pairs of inter-stimulus intervals (ISIs) on the induction of LTP. Therefore, the function of frequency (first-order) and temporal pattern (second-order) was examined using this imaging technique. To investigate the dependence of LTP on frequency, periodic stimuli with the same number of pulses were applied at different frequencies (1-10 Hz, n=200) to Schaffer commissural-collateral fibers. While stimulus frequencies from 2-10 Hz induced LTP of varying magnitudes and low-frequency stimuli (1 Hz) induced long-term depression (LTD), spatial distribution remained consistent. These results suggest that induction frequency has a greater effect on the magnitude of LTP than on its spatial distribution. By employing nonperiodic stimuli at the same mean frequency (2 Hz), the effect of varying the temporal structure of a stimulus was also investigated. As the correlation of successive ISIs was increased from negative to positive, not only did the magnitude of LTP increase, there was also a statistically significant change in the spatial distribution of LTP. Interestingly, when a strong negatively correlated stimulus was applied, both LTP and LTD were simultaneously observed in the CA1 area. It was also found that the magnitude of LTP 200-300 mum distal to the cellular layer was larger than that of the LTP induced proximal (<100 microm) to that layer. These results support the hypothesis that the spatial distribution of LTP throughout the hippocampus relies principally on the temporal patterning of input stimulation. This insight into the structure of the CA1 neural network may reveal the importance of stimulus timing events in the spatial encoding of memories.

Action Potentials↗

Method for quantitative determination of spatial polymer distribution in alginate beads using Raman spectroscopy.

A new method based on Raman spectroscopy is presented for non-invasive, quantitative determination of the spatial polymer distribution in alginate beads of approximately 4 mm diameter. With the experimental setup, a two-dimensional image is created along a thin measuring line through the bead comprising one spatial and one spectral dimension. For quantitative analysis of the Raman spectra, the method of indirect hard modeling was applied to make use of the information contained in the entire recorded spectra. For quantification of the alginate signals from within the beads, a calibration curve acquired from sodium alginate solutions was used after it was shown that only negligible differences occur between signals from alginate solutions and alginate gels. The distribution of alginate over the bead gel matrix was acquired with high spatial (51 microm) and time (12 s) resolution. The inhomogeneous distribution obtained using the new measuring technique is qualitatively in excellent agreement with data from the literature. In contrast to known measuring techniques, correct quantitative information about the spatial polymer distribution within the matrix was derived. It gave an alginate mass fraction of approximately 0.045 g/g at the edges and 0.02 g/g in the center of the beads. Next to the determination of mere polymer concentrations, the excellent time resolution of the presented method will enable investigation of the dynamic process of gel formation and it will also serve as a basis for investigation of mass transfer of small diffusing molecules in alginate matrices.

Alginates↗

Spatial indoor radon distribution in Mexico City.

We present a spatial analysis of residential radon concentrations in the Mexico City Metropolitan Area, which we intend to use to assign radon exposure in an ongoing case-control study. As part of a probabilistic household survey, carried out between May and June 1999, 501 dwellings were selected for indoor placement of solid state nuclear track detectors (LR 115) in a cup array over a period of approximately 90 days. As part of the sampling design, the city was grid partitioned into nine zones and a sample of dwellings was selected in each zone. All zones were simultaneously surveyed. The stratified sampling design allowed us to obtain radon geometric means, adjusted for household characteristics, week of detector placement and number of days of measurement for these zones. Additionally, adjusted geometric means were estimated for the 100 census tracts surveyed and this information was used to obtain a more detailed spatial distribution of residential radon levels through kriging interpolation and surface contouring. Radon levels depended on the room of placement, the floor level and the ventilation habits but not on building materials. Regarding the city zone, the highest adjusted geometric mean was found in the southwest (136 Bqm(-3)), where 46% of the households had an estimated radon level in excess of 200 Bqm(-3). In the rest of the city, the geometric mean concentration ranged between 41 and 98 Bqm(-3). A more detailed spatial distribution showed that, in general, most of the eastern and middle zones of the city had estimated radon geometric means below 74 Bqm(-3), while the western ones had geometric means above this concentration. Very high geometric means, exceeding 111 Bqm(-3) and even reaching 288 Bqm(-3), are estimated for some areas located in the southern and western zones of Mexico City. The obtained spatial distribution shows that the areas with very high estimated residential radon concentrations are close to inactive volcanic mountains. We believe that the geo-statistical techniques, we have used, offer reasonably good estimates of the average spatial residential radon distribution in Mexico City under average ventilation in homes. The use of this indirect approach for radon exposure measurement in epidemiological studies is an inexpensive alternative to direct radon exposure measurement but may be subject to non-differential misclassification error. The effect of such error on the detection of a real increase in lung cancer risk from indoor radon remains to be determined.

Air Pollution, Indoor↗

Calcium oscillations in a triplet of pancreatic acinar cells.

We use a mathematical model of calcium dynamics in pancreatic acinar cells to investigate calcium oscillations in a ring of three coupled cells. A connected group of cells is modeled in two different ways: 1), as coupled point oscillators, each oscillator being described by a spatially homogeneous model; and 2), as spatially distributed cells coupled along their common boundaries by gap-junctional diffusion of inositol trisphosphate and/or calcium. We show that, although the point-oscillator model gives a reasonably accurate general picture, the behavior of the spatially distributed cells cannot always be predicted from the simpler analysis; spatially distributed diffusion and cell geometry both play important roles in determining behavior. In particular, oscillations in which two cells are in synchrony, with the third phase-locked but not synchronous, appears to be more dominant in the spatially distributed model than in the point-oscillator model. In both types of model, intercellular coupling leads to a variety of synchronous, phase-locked, or asynchronous behaviors. For some parameter values there are multiple, simultaneous stable types of oscillation. We predict 1), that intercellular calcium diffusion is necessary and sufficient to coordinate the responses in neighboring cells; 2), that the function of intercellular inositol trisphosphate diffusion is to smooth out any concentration differences between the cells, thus making it easier for the diffusion of calcium to synchronize the oscillations; 3), that groups of coupled cells will tend to respond in a clumped manner, with groups of synchronized cells, rather than with regular phase-locked periodic intercellular waves; and 4), that enzyme secretion is maximized by the presence of a pacemaker cell in each cluster which drives the other cells at a frequency greater than their intrinsic frequency.

Animals↗

In vivo measurement of spatial dose distribution with thermoluminescent sheet around high dose-rate intracavitary source: application to rectal cancer.

For intracavitary high dose-rate radiation therapy, a thermoluminescent [TL] sheet for in vivo measurement of spatial dose distribution around source has been recently developed. The TL sheet was found to have a linear response with a very wide dynamic range from at least 0.002 cGy to 5000 cGy for 60Co gamma-rays. This TL sheet (40 cm x 50 cm x 200 microns), which is composed of Teflon mixed with BaSO4:Eu doped powder, is very flexible and can be cut to the desired size. In addition, this sheet is easy to handle because of its insensitivity to room light. The spatial dose distribution is displayed in a color mode by using a newly developed TL sheet readout system. For a clinical application, the TL sheet was wrapped on an applicator for intracavitary radiation therapy of a rectal cancer and was inserted into the rectum. The location of the TL sheet could be confirmed with diagnostic X ray film. After irradiation with high dose-rate 60Co source, the in vivo relative dose distribution on the surface of the rectum was determined. This TL sheet provided a convenient means of measuring the relative dose distributions around 60Co sources of various patterns in intracavitary radiation therapy.

Brachytherapy↗

Retinal distribution of attention.

Spatial distribution of attention around a fixation point was investigated by comparing cued vs noncued conditions. The curve of retinal locations for the cued condition was flatter than that of the noncued condition, indicating attentional spotlight is focused around the fixation point.

Attention↗

Modulation of repolarization by electrotonic interactions.

The expression of repolarization properties in different regions of the ventricle is a complex interaction between the spatial distribution of intrinsic properties of the cells and the spatial distribution of the coupling resistance among the cells of the cardiac syncytium. Our theoretical studies show that electrotonic modulation of repolarization among cells with intrinsically different repolarization properties can make the expressed properties of cells at a given location quite different from the intrinsic properties of these cells. These interactions are simulated both for a simple two cell system as well as for more complex geometries, showing that the spatial distribution of expressed repolarization properties is quite extensive, and, more specifically, is much more extensive than would be expected by passive electrotonic interactions.

Action Potentials↗

Distribution of single-channel conductances in cultured rat hippocampal neurons.

1. The nonhomogeneous spatial distribution of ionic channels in neurons has been implied from intracellular recordings at somatic and dendritic locations. These reports indicate that Na- and Ca-dependent regenerative currents are distributed differently throughout the neuron. Although a variety of K conductances and a noninactivating Na conductance have been described in intracellular studies, little is known about the spatial distribution of inward and outward currents throughout different regions of the neuron. 2. We recorded from cell-attached patches from cultured hippocampal cells from 1-day-old rats. The cells were cultured for 3-21 days. The spatial distribution of a variety of ionic channels was determined by comparing the conductances from somatic and dendritic membranes. Single-channel currents obtained from cell-attached patches were identified by the time course of ensemble (averaged) responses, voltage dependence, and the effect of channel blocking agents. 3. We consistently observed that only the rapidly inactivating inward current was localized to the soma. The other channel types that we studied, including an inward noninactivating, delayed rectifier and transient A-type currents, were observed in both the somatic and dendritic regions. 4. We suggest that the distribution of ionic conductances that we have observed may be functional in limiting excitability during development of neurons.

Animals↗

Automated extraction and variability analysis of sulcal neuroanatomy.

Systematic mapping of the variability in cortical sulcal anatomy is an area of increasing interest which presents numerous methodological challenges. To address these issues, we have implemented sulcal extraction and assisted labeling (SEAL) to automatically extract the two-dimensional (2-D) surface ribbons that represent the median axis of cerebral sulci and to neuroanatomically label these entities. To encode the extracted three-dimensional (3-D) cortical sulcal schematic topography (CSST) we define a relational graph structure composed of two main features: vertices (representing sulci) and arcs (representing the relationships between sulci). Vertices contain a parametric representation of the surface ribbon buried within the sulcus. Points on this surface are expressed in stereotaxic coordinates (i.e., with respect to a standardized brain coordinate system). For each of these vertices, we store length, depth, and orientation as well as anatomical attributes (e.g., hemisphere, lobe, sulcus type, etc.). Each arc stores the 3-D location of the junction between sulci as well as a list of its connecting sulci. Sulcal labeling is performed semiautomatically by selecting a sulcal entity in the CSST and selecting from a menu of candidate sulcus names. In order to help the user in the labeling task, the menu is restricted to the most likely candidates by using priors for the expected sulcal spatial distribution. These priors, i.e., sulcal probabilistic maps, were created from the spatial distribution of 34 sulci traced manually on 36 different subjects. Given these spatial probability maps, the user is provided with the likelihood that the selected entity belongs to a particular sulcus. The cortical structure representation obtained by SEAL is suitable to extract statistical information about both the spatial and the structural composition of the cerebral cortical topography. This methodology allows for the iterative construction of a successively more complete statistical models of the cerebral topography containing spatial distributions of the most important structures, their morphometrics, and their structural components.

Algorithms↗

Calcium concentration dynamics produced by synaptic activation of CA1 hippocampal pyramidal cells.

The spatial and temporal dynamics of many electrophysiological and biochemical processes in nerve cells are in turn dependent on the concentration dynamics of the second messenger calcium. We have used microfluorimetry of the calcium indicator fura-2 (Grynkiewicz et al., 1985) to measure and characterize synaptically activated calcium changes in individual CA1 pyramidal cells contained within guinea pig hippocampal slices. One component of the calcium changes was largely produced by influx through voltage-dependent Ca2+ channels (VDCCs). It consisted of large transient accumulations in the proximal-apical and basal dendrites; the amplitude was smaller in the distal-apical dendrites and the soma. This spatial profile was insensitive to the method of cell activation: stimulation of inputs located at different positions on the dendritic tree as well as antidromic stimulation produced only slight modifications. This component was not blocked by the NMDA antagonist 5-amino-4-phosphonovalerate (AP5) (Collingridge et al., 1983), was greatly reduced by Cd2+, partially reduced by nifedipine, and was increased by Bay-K 8644, providing the evidence that it was largely produced by influx through VDCCs. Blocking postsynaptic Na+ channels with QX-314 greatly reduced the accumulation amplitude, and spatial differences between proximal-dendritic and distal-dendritic regions were less pronounced, suggesting that active sodium conductances contribute significantly to the spatial activation of calcium conductances. Residual spatial differences that persist in QX-314 experiments are consistent with the idea that VDCCs have decreased density on distal-apical dendrites. A second component of accumulation was induced by ionic currents through NMDA receptor channels. It was blocked by AP5, unaffected by QX-314, attenuated and slowed down by elevated calcium buffering, and spatially localized to regions receiving activated synaptic inputs. The magnitude of this component was strongly dependent on the frequency and amplitude of synaptic activation. At high frequency, it was generally very large, often saturating the fura-2 (> 2 microM). Measurements made with the indicator furaptra also showed large localized AP5-sensitive fluorescence changes. Our results suggest that in dendritic regions near activated input fibers calcium levels may reach 2-10 microM. In general, our measurements of calcium dynamics provide an experimental basis for evaluating the spatial distribution of calcium conductances, the spatial distribution of calcium-activated electrophysiological and biochemical processes, and the spatial uniformity of calcium buffering and removal systems in CA1 hippocampal pyramidal cells. The time course and amplitude of Ca2+ transients we measured suggest that activation of Ca(2+)-dependent conductances [e.g., IK(Ca)] will be markedly different for different cellular regions.(ABSTRACT TRUNCATED AT 400 WORDS)

2-Amino-5-phosphonovalerate↗

Discrimination training alters object representations in human extrastriate cortex.

Visual object recognition relies critically on learning. However, little is known about the effect of object learning in human visual cortex, and in particular how the spatial distribution of training effects relates to the distribution of object and face selectivity across the cortex before training. We scanned human subjects with high-resolution functional magnetic resonance imaging (fMRI) while they viewed novel object classes, both before and after extensive training to discriminate between exemplars within one of these object classes. Training increased the strength of the response in visual cortex to trained objects compared with untrained objects. However, training did not simply induce a uniform increase in the response to trained objects: the magnitude of this training effect varied substantially across subregions of extrastriate cortex, with some showing a twofold increase in response to trained objects and others (including the right fusiform face area) showing no significant effect of training. Furthermore, the spatial distribution of training effects could not be predicted from the spatial distribution of either pretrained responses or face selectivity. Instead, training changed the spatial distribution of activity across the cortex. These findings support a dynamic view of the ventral visual pathway in which the cortical representation of an object category is continuously modulated by experience.

Discrimination, Psychological↗

Vertical and horizontal visual whole-field motion differently affect the metabolic activity of the rat medial terminal nucleus.

The metabolic activity of the medial terminal nucleus (MTN) of the Accessory Optic System was studied by means of the [14C]2-deoxyglucose (2-DG) method in Long-Evans rats exposed to moving and stationary visual stimuli. In particular we explored the rate of local cerebral glucose utilization (LCGU) and the spatial distribution of 2-DG uptake within MTN related to visual stimuli capable of triggering optokinetic nystagmus. It was found that increases in MTN metabolism accompanied the retinal slip signals evoked by whole-field visual patterns moving in the vertical as well as in the horizontal direction. At the same level of luminous flux neither the same but stationary pattern, nor constant, diffuse illumination were able to elicit comparable changes in MTN metabolic rates. The effects of vertical and horizontal motions differed, however, from each other. In binocular testing LCGU rates resulted significantly higher after vertically moving patterns and upon the same stimulus condition the spatial distribution of 2-DG matched very closely the spatial distribution of the retinal afferents and the cellular density within MTN, in sharp contrast with the diffuse spreading out of the label across the nucleus following horizontal motion. In monocular testings only the vertically moving patterns were able to increase LCGU rates significantly and then in contralateral MTN alone. However, comparison between the levels of glucose consumption measured in binocular and in monocular vision also showed the involvement of the uncrossed retinal path in relaying the retinal slip signals to MTN. No difference in LCGU and in spatial distribution of the label were finally observed in relation to the upward or to the downward direction of the moving pattern.

Animals↗

Spatial functional distribution in the corticospinal tract at the corona radiata: a three-dimensional anisotropy contrast study.

The spatial functional distribution of the nerve fibers was investigated in the corticospinal tract at the level of the corona radiata. Thirteen patients with corona radiata infarction underwent axial single-shot echo planar diffusion-weighted magnetic resonance imaging using a 1.5 Tesla scanner. Image analysis used the three-dimensional anisotropy contrast (3DAC) method to demarcate the nerve fibers in the corticospinal tract. Axial 3DAC images demonstrated the corticospinal tract as a distinct area indicating nerve fiber integrity in all normal hemispheres and infarction as a dark or black area in affected hemispheres. Seven patients with upper extremity-dominant motor dysfunction had infarction located in the middle one third of the corticospinal tract. A patient with lower extremity-dominant motor dysfunction had infarction in the posterior one third. Five patients with equal motor dysfunction in the upper and lower extremities had infarction in both the middle and posterior one thirds of the corticospinal tract. The recovery of motor dysfunction at one month follow up correlated with the location of the corticospinal tract injury on the initial 3DAC images. The findings of the 3DAC images provide an indicator of the pattern and the recovery from acute and chronic motor dysfunction in patients with corona radiata infarction.

Aged↗