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A multiresolution image based approach for correction of partial volume effects in emission tomography.

Partial volume effects (PVEs) are consequences of the limited spatial resolution in emission tomography. They lead to a loss of signal in tissues of size similar to the point spread function and induce activity spillover between regions. Although PVE can be corrected for by using algorithms that provide the correct radioactivity concentration in a series of regions of interest (ROIs), so far little attention has been given to the possibility of creating improved images as a result of PVE correction. Potential advantages of PVE-corrected images include the ability to accurately delineate functional volumes as well as improving tumour-to-background ratio, resulting in an associated improvement in the analysis of response to therapy studies and diagnostic examinations, respectively. The objective of our study was therefore to develop a methodology for PVE correction not only to enable the accurate recuperation of activity concentrations, but also to generate PVE-corrected images. In the multiresolution analysis that we define here, details of a high-resolution image H (MRI or CT) are extracted, transformed and integrated in a low-resolution image L (PET or SPECT). A discrete wavelet transform of both H and L images is performed by using the "à trous" algorithm, which allows the spatial frequencies (details, edges, textures) to be obtained easily at a level of resolution common to H and L. A model is then inferred to build the lacking details of L from the high-frequency details in H. The process was successfully tested on synthetic and simulated data, proving the ability to obtain accurately corrected images. Quantitative PVE correction was found to be comparable with a method considered as a reference but limited to ROI analyses. Visual improvement and quantitative correction were also obtained in two examples of clinical images, the first using a combined PET/CT scanner with a lymphoma patient and the second using a FDG brain PET and corresponding T1-weighted MRI in an epileptic patient.

Algorithms↗

A biodegradable hybrid sponge nested with collagen microsponges.

A biodegradable hybrid sponge of poly(DL-lactic-co-glycolic acid) (PLGA) and collagen was fabricated by forming microsponges of collagen in the pores of PLGA sponge. Observation of the PLGA-collagen hybrid sponge by scanning electron microscopy (SEM) showed that microsponges of collagen with interconnected pore structures were formed in the pores of PLGA sponge. The hybrid structure further was confirmed by scanning electron microscopy-electron probe microanalysis (SEM-EPMA), and elemental nitrogen was detected in the microsponges of collagen and on the pore surfaces of PLGA, but not in cross-sections of PLGA regions. The formation of collagen microsponges was dependent on collagen concentration, the effective range of which was from 0.1 to 1.5 (w/v) %. The mechanical strength of the hybrid sponge was higher than that of either PLGA or collagen sponges, in both dry and wet states. The wettability with water was improved by hybridization with collagen, which facilitated cell seeding in the hybrid sponge. Mouse fibroblast L929 cells attached well and spread on the surfaces of the microsponges of collagen in the hybrid sponge. The distribution of cells was spatially uniform throughout the hybrid sponge. Use of the PLGA sponge as a skeleton facilitated formation of the hybrid sponge into desired shapes with high mechanical strength while collagen microsponges contributed good cell interaction and hydrophilicity.

Animals↗

Self-calibrated spiral SENSE.

Current standard sensitivity-encoded parallel imaging (SENSE) utilizes a fully sampled low-resolution reference scan to estimate the coil sensitivities. This reference scan adds scan time and may introduce misregistration artifacts. The purpose of this study was to investigate the feasibility of estimating the coil sensitivities for spiral SENSE directly from an undersampled k-space center. The limited spatial frequencies of the coil sensitivities, and the undersampling beyond the Nyquist radius cause image artifacts. A point spread function (PSF) analysis and experiments on both phantoms and humans identified an optimal radius for the k-space center by minimizing these image artifacts. The preliminary data indicate that self-calibrated SENSE is as accurate as standard SENSE, which uses a fully sampled reference scan.

Algorithms↗

Individual and multi-environment combined analyses identify QTLs for morphogenetic and reproductive development traits in white clover (Trifolium repens L.).

White clover (Trifolium repens L.) is a key component legume of temperate pasture agriculture and an important target for molecular marker-assisted plant breeding. A genetic map of white clover has been used to assess genetic control of agronomically important traits that vary in the F2(I.4RxI.5J) mapping family. Phenotypic analysis was performed for a range of vegetative morphogenesis traits (such as leaf area, internode length, plant height and plant spread) and reproductive morphogenesis and development traits (such as flowering date, floral intensity and seed yield), with both spatial and temporal replication. A multi-environment combined analysis (combined analysis) has been performed for traits assessed across multiple experimental datasets in order to identify consistent genetic effects. Quantitative trait locus (QTLs) were detected for the majority of traits, and the locations and magnitudes of QTL effects were compared between individual and combined analyses. This molecular genetic dissection of agronomic traits in white clover provides the basis for equivalent studies in more complex populations, design of marker-assisted selection strategies and comparative genetics with model legume species. Selection for QTLs derived from the combined analysis will permit robust improvement of phenotypic traits over different environments.

Chromosome Mapping↗

Patch-clamp recording from Müller (glial) cell endfeet in the intact isolated retina and acutely isolated Müller cells of mouse and guinea-pig.

Müller cells span through the entire retina and terminate with the formation of endfeet at the vitreous body. These endfeet are thought to be specialized for maintaining the K+ homeostasis in the retina based on the assumption that voltage signals can passively spread from the cell body to the endfeet. We employed the patch-clamp technique to study the physiological properties of these endfeet in a retinal wholemount preparation from guinea-pig or mouse. After assessing one endfoot with the patch pipette and establishing the whole cell recording configuration, a membrane area which approximately matched the size of one endfoot and proximal process could be voltage-clamped. This morphological correlation could be established by filling the cytoplasm with the fluorescent dye Lucifer Yellow via the patch-pipette. The morphological, immunocytochemical and ultrastructural inspection of the recorded cells revealed that mouse Müller cell endfeet were connected by only a thin stalk to the proximal process. In contrast, guinea-pig endfeet were connected by thick stalks. The endfoot current in the mouse was dominated by a voltage and time-independent K+ conductance. In contrast, in some of the recordings from guinea-pig, delayed and inwardly rectifying K+ currents were observed. These voltage-gated currents were more frequently observed or were facilitated when the membrane area under voltage clamp was increased, blocking the passive K+ currents by Ba2+ in both, mouse and guinea-pig. We thus assume that the voltage-gated currents were not in the endfeet membrane, but rather in the proximal process and could thus be better activated in the guinea-pig with its thicker stalk or after increasing the membrane area under voltage clamp control. Similar results were obtained in freshly isolated Müller cells; in contrast to the cells from the wholemount the voltage-gated currents were more frequently observed. These studies demonstrate that the Müller cell endfoot of the mouse with its vascularized retina is an electrically isolated unit and that voltage signals do not spread to the proximal process. Such a property would, however, be required for the redistribution of K+ via spatial buffer currents. In contrast, guinea-pig Müller glial cells with their stout morphological connection between endfoot and proximal process are better suited to fulfil this task.

Animals↗

Photoreceptor coupling and boundary detection.

Electrical coupling between photoreceptors results in the extensive spreading of output potentials along the syncytium of photoreceptor terminals. This smoothing of output potentials seems to make spatial resolution worse. However, the photoreceptor noise that is considered to be non-correlated both in space and time is smoothed to the greater extent than the correlated potential difference across the boundaries between areas of different brightness. This improves the signal-to-noise ratio more for more extended boundaries and favours lowering the threshold so that they can be detected more easily during the subsequent processing. The results have a striking parallel with a well known dependence of contrast threshold on stimulus size as measured psychophysically.

Contrast Sensitivity↗

Coupling and uncoupling of amphibian neuroglia.

Glial cells in the optic nerve of Necturus are coupled to each other by low resistance pathways which also permit the diffusion of the fluorescent dye Lucifer Yellow CH among the cells. The spread of dye is readily observed as nuclei of cells distant from the site of intracellular injection are stained. By contrast, horseradish peroxidase does not traverse the intercellular pathways. This protein remains in the injected cell. The addition of weak acids (carbonic or propionic) to the bathing medium reversibly uncouples the glia; it blocks the spread of ionic current and Lucifer Yellow among the cells. A block of ionic coupling will block the spatial buffering of potassium by the glial syncytium.

Animals↗

Influence of hole shape on collimator performance.

The imaging properties of triangular and hexagonal hole shapes were compared for low energy scintillation camera collimators. The response of both hole shapes was calculated using a ray-tracing computer program, corresponding collimators were constructed and their performance was evaluated by measuring line spread functions and imaging bar patterns. The triangular hole shape was found to give better results above a spatial frequency of 1-5 cycles cm-1, below that frequency the hexagonal hole shape is superior. The differences, however, are small and the superior performance of the hexagonal hole collimator at clinically significant spatial frequencies is barely visible in bar pattern images.

Computers↗

Imaging using parallel integrals in optical projection tomography.

We develop and demonstrate improved image-forming optics for optical projection tomography (OPT), with which the parallel integral throughout an object can be obtained. This method results in an improved resolution for OPT images, especially for the cross sections far from the optical axis of the image-forming optics. We find the optimal configuration used in our OPT system by use of a point spread function and simulation technique. The new method is validated by both numerical simulations and experimental results. The spatial resolution of the OPT system presented is approximately 40 microm.

Animals↗

Laminar analysis of cerebral blood flow in cortex of rats by laser-Doppler flowmetry: a pilot study.

Laser-Doppler flowmetry (LDF) is a reliable method for estimation of relative changes of CBF. The measurement depth depends on wavelength of the laser light and the separation distance of transmitting and recording optical fibers. We designed an LDF probe using two wavelengths of laser light (543 nm and 780 nm), and three separation distances of optical fibers to measure CBF in four layers of the cerebral cortex at the same time. In vitro comparison with electromagnetic flow measurements showed linear relationship between LDF and blood flow velocity at four depths within the range relevant to physiologic measurements. Using artificial brain tissue slices we showed that the signal for each channel decreased in a theoretically predictable fashion as a function of slice thickness. Application of adenosine at various depths in neocortex of halothane-anesthetized rats showed a predominant CBF increase at the level of application. Electrical stimulation at the surface of the cerebellar cortex demonstrated superficial predominance of increased CBF as predicted from the distribution of neuronal activity. In the cerebellum, hypercapnia increased CBF in a heterogeneous fashion, the major increase being at apparent depths of approximately 300 and 600 microns, whereas in the cerebral cortex, hypercapnia induced a uniform increase. In contrast, the CBF response to cortical spreading depression in the cerebral cortex was markedly heterogeneous. Thus, real-time laminar analysis of CBF with spatial resolution of 200 to 300 microns may be achieved by LDF. The real-time in depth resolution may give insight into the functional organization of the cortical microcirculation and adaptive features of CBF regulation in response to physiologic and pathophysiologic stimuli.

Animals↗

Spatial frequency thresholds versus border enhancement: sensitivity to retinal defocus.

Two psychophysical methods for monitoring retinal defocus are compared. One methods, spatial frequency thresholds with sine wave grids, is well known. The second methods is relatively new: it uses the effect of defocus on the width of the perceptually enhanced region of brightness or darkness generated next to a luminance edge. Both methods produce characteristic functions as the retinal image is thrown out of focus. However, spatial frequency thresholds appear relatively unchanged until a certain level of defocus has been introduced, whereas border enhancement spread is affected even by very small deviations from focus. It is suggested that variations in the frequency threshold are dampened in the near-focus range by aberrations and by the nonlinear relation between contrast and defocus, but chiefly by the resolution limit of the retina and visual pathways. However, it is found that the apparent lack of response to small deviations from focus is due to a drastic compression rather than absence of threshold changes. If responses through this region are shown on a greatly expanded scale in relation to the rest of the function. fluctuations appear that often match those generated by border enhancement.

Differential Threshold↗

Localization-delocalization transition in quantum dots

Single-electron capacitance spectroscopy precisely measures the energies required to add individual electrons to a quantum dot. The spatial extent of electronic wave functions is probed by investigating the dependence of these energies on changes in the dot confining potential. For low electron densities, electrons occupy distinct spatial sites localized within the dot. At higher densities, the electrons become delocalized, and all wave functions are spread over the full dot area. Near the delocalization transition, the last remaining localized states exist at the perimeter of the dot. Unexpectedly, these electrons appear to bind with electrons in the dot center.

Journal Article↗

5-Hydroxytryptophan modulates postprandial motor patterns of canine proximal small intestine.

The aim of the study was to clarify whether 5-hydroxytryptophan (5-HTP) stimulates the postprandial motor pattern of the duodenum in a similar way as that of the adjacent jejunal segment in dogs. Computerized analysis of motor patterns recorded by closely spaced strain gauges focused on the temporal and spatial distribution of the contractions. Results indicate that 5-HTP increased the incidence and the length of the spread of contraction waves after both an acaloric and a nutrient meal in the duodenum as well as in the adjacent jejunal segment. Effects were more pronounced after the nutrient than after the acaloric meal. After the nutrient meal, but not after the acaloric meal, 5-HTP additionally enhanced the number of both duodenal and jejunal contractions per minute and increased the force of duodenal contractions. The acaloric meal induced significant differences in the motor patterns between the duodenum and the adjacent jejunum. 5-HTP abolished these differences owing to a relatively stronger stimulation of duodenal motility. 5-HTP did not affect gastric emptying of both meals. We conclude (i) that 5-HTP is a potent stimulator of propagated contractions both in the duodenum and the adjacent jejunal segment and (ii) that intestinal motor patterns can be regulated independently of gastric emptying.

5-Hydroxytryptophan↗

Extended resolution wide-field optical imaging: objective-launched standing-wave total internal reflection fluorescence microscopy.

Standing-wave total-internal-reflection fluorescence (SW-TIRF) microscopy uses a super-diffraction-limited standing evanescent wave to extract the high-spatial-frequency content of an object through a diffraction-limited optical imaging system. The effective point-spread function is better than a quarter of the emission wavelength. With a 1.45 numerical aperture objective and 532 nm excitation wavelength, a Rayleigh resolution of approximately 100 nm can be achieved, which is better than twice the resolution of conventional TIRF microscopy. This first experimental realization of SW-TIRF in an objective-launched geometry demonstrates the potential for extended resolution imaging at high speed by using wide-field microscopy.

Equipment Design↗

Analysis of depth profiling data obtained by confocal Raman microspectroscopy.

The nominal depth resolution achieved in confocal Raman microscopy is on the order of a few micrometers. Often, however, the depth resolution is decreased by light refraction at the sample surface. The problem can be avoided with the use of an immersion objective and index matching oils. Through this intervention the instrument point-spread function (PSF) can be assumed to be independent of the depth of focus in the sample, and spatially invariant depth profiles can be acquired. In this work the instrument PSF was determined by measuring a depth profile of a thick uniform sample and calculating the first derivative of the depth profile curve. The first-derivative method was also used to determine sample thickness. Convolution with the PSF makes it possible to simulate the behavior of the instrument with different sample functions. It is also possible to use the instrument PSF to deconvolve depth-profiling data. Deconvolution reduces the blurring effect of the instrument and increases the depth resolution. Deconvolution can also be used in analysis of the sample surface position and in layer structure analysis. In this paper we show how the convolution integral can be used with the immersion sampling technique to determine the PSF and how the sample thickness can be determined.

Algorithms↗

Advanced tracer techniques to monitor synaptic activity.

The two approaches presented here bypass postsynaptic receptors as indicators of quantal release, and thus they can provide information which is clearly distinct from that obtained with standard electrophysiological techniques. Indeed, the inherently variable responsiveness of the postsynaptic membrane makes it an unreliable indicator of presynaptic activity and this has fueled a lot of controversy, particularly in the area of synaptic plasticity. A major advantage of these two methods is their ability to detect changes at the single bouton level. This offers a lot of advantages including the possibility to study the functional role for exo-endocytosis but also plasticity against a background of great variability among a large number of synapses. The spatial resolving power of FM1-43 and anti-synaptotagmin antibodies may be valuable in future studies of spread of LTP between neighboring synapses and in the mapping the pattern of neuronal activity in complex networks of neurons.

Animals↗

Ontogenetic aspects of dental evolution.

The evolution of dental ontogeny in the vertebrates is reviewed. Teeth probably originated as dermal structures, which secondarily spread to the mouth, where they became associated with bones. Tooth formation is a repetitive process, resulting in spatially separate units, and primitively it continued throughout life. Development of conical teeth commences at the tip and extends basally; folding of the basal lamina of the inner dental epithelium results in complex shapes, as in mammalian molars. Heterodonty, the divergent development of the teeth in a dentition, has evolved in a number of vertebrates, particularly mammals. Experimental analysis of dental development is still at an early stage, and the explanation of evolutionary changes in developmental terms is largely speculative Mammals are atypical vertebrates in many ways, and more studies of lower vertebrates, especially fishes, are needed.

Animals↗

Neuronal control of development in hydra.

Hydra is an excellent model system for developmental biology, because pattern formation processes can be easily studied in regeneration, transplantation, and reaggregation experiments. At the cellular level hydra has the advantage that it contains only a few basic cell types and that differentiation pathways are short. Two types of signals, produced and released by nerve cells, control the spatial and temporal patterns of differentiation. Positive signals induce specific local differentiation events, and negative signals inhibit the spread of such inductions to larger areas. Head-specific growth and differentiation are controlled by head activator and head inhibitor, food-specific processes are regulated by foot activator and foot inhibitor. The activators are peptides, the inhibitors are low-molecular-weight substances. The sequence of the head activator is known, and it is conserved throughout the animal kingdom. At the cellular level head activator exerts three types of effects in hydra. It stimulates cells to divide, and it is responsible for the determination and the final differentiation of nerve cells and head-specific epithelial cells. For nerve-cell differentiation the cAMP pathway is used as second messenger system. Components of this pathway were identified in hydra. In mammals head activator is produced by nerve and neuro-endocrine cells, and it acts as mitogen on cells of neural origin. It is present early in neural development and in abnormal neural development, such as brain and neuroendocrine tumours.

Amino Acid Sequence↗