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Phantoms for IMRT dose distribution measurement and treatment verification.

BACKGROUND: The verification of intensity-modulated radiation therapy (IMRT) patient treatment dose distributions is currently based on custom-built or modified dose measurement phantoms. The only commercially available IMRT treatment planning and delivery system (Peacock, NOMOS Corp.) is supplied with a film phantom that allows accurate spatial localization of the dose distribution using radiographic film. However, measurements using other dosimeters are necessary for the thorough verification of IMRT. METHODS: We have developed a phantom to enable dose measurements using a cylindrical ionization chamber and the localization of prescription isodose curves using a matrix of thermoluminescent dosimetry (TLD) chips. The external phantom cross-section is identical to that of the commercial phantom, to allow direct comparisons of measurements. A supplementary phantom has been fabricated to verify the IMRT dose distributions for pelvis treatments. RESULTS: To date, this phantom has been used for the verification of IMRT dose distributions for head and neck and prostate cancer treatments. Designs are also presented for a phantom insert to be used with polymerizing gels (e.g., BANG-2) to obtain volumetric dose distribution measurements. CONCLUSION: The phantoms have proven useful in the quantitative evaluation of IMRT treatments.

Head and Neck Neoplasms↗

[Multiple correlative studies of stereotaxic biopsies of brain tumors].

The opportunity of having several samples at the same site which could be spatially localized allows an intensive exploitation of stereotactic biopsies of brain tumors: the pathological data may be correlated to other measures, performed at the same site (electrical impedance X ray absorption coefficient) or on other samples (NMR relaxation times, water content, nucleic acids). These samples are available for oncology experiments in cellular biology (cell cultures, grafts on nude mice) or in molecular biology (DNA and RNA hybridization with specific nucleic acid probes). We were therefore able: 1) to study the diagnostic homologies between pathology and histology examinations; 2) to show that T1 and T2 NMR relaxation times are 2 times longer in tumor tissues than in normal brain; 3) to show that the electrical impedance is decreased by a factor 2 in brain tumors; 4) to show the absence of integrated viral genomic sequences and the existence of oncogenes association patterns in brain tumors by hybridization of specific sequences; 5) to establish permanent cell lines, the tumorigenicity of which is assayed by grafting on nude mice. Therefore, stereotactic biopsies appear to be, provided they are intensively and rationally exploited, a major research tool in an area which remains unsensitive to the various therapeutic approaches.

Animals↗

Chronic microstimulation in the feline ventral cochlear nucleus: physiologic and histologic effects.

This study was conducted to help to establish the feasibility of a multi-channel auditory prosthesis based on microstimulation within the human ventral cochlear nucleus, and to define the range of stimulus parameters that can be used safely with such a device. We chronically implanted activated iridium microelectrodes into the feline ventral cochlear nucleus and, beginning 80-250 days after implantation, they were pulsed for 7 h/day, on up to 21 successive days. The stimulus was charge-balanced pulses whose amplitude was modulated by a simulated human voice. The pulse rate (250 Hz/electrode) and the maximum pulse amplitude were selected as those that are likely to provide a patient with useful auditory percepts. The changes in neuronal responses during the multi-day stimulation regimens were partitioned into long-lasting, stimulation-induced depression of neuronal excitability (SIDNE), and short-acting neuronal refractivity (SANR). Both SIDNE and SANR were quantified from the changes in the growth functions of the evoked potentials recorded in the inferior colliculus. All of the stimulation regimens that we tested induced measurable SIDNE and SANR. The combined effect of SIDNE and the superimposed SANR is to depress the neuronal response near threshold, and thereby, to depress the population response over the entire amplitude range of the stimulus pulses. SIDNE and SANR may cause the greatest degradation of the performance of a clinical device at the low end of the amplitude range, and this may represent an inherent limitation of this type of spatially localized, high-rate neuronal stimulation. We determined sets of stimulus parameters which preserved most of the dynamic range of the neuronal response, when using either long (150 micros/phase) or short (40 micros/phase) stimulus pulses. Increasing the amplitude of the stimulus was relatively ineffective as a means of increasing the dynamic range of neuronal response, since the greater stimulus amplitude induced more SIDNE. All of the pulsed and unpulsed electrode sites were examined histologically, and no neuronal changes attributable to the stimulation were detected. There was some aggregation of glial cells immediately adjacent to some of the electrodes that were pulsed with the short-duration pulses, and at the highest current densities.

Animals↗

Adaptive data acquisition in MRI.

We propose an adaptive data acquisition technique that depends on the object to be imaged in magnetic resonance (MR) imaging. In this paper, we employed a matching pursuit (MP) algorithm to achieve the adaptive data acquisition. Since the matching pursuit is a greedy algorithm to find RF and gradient waveforms which are the best match for an object-signal, the signal can be decomposed with a few iterations and thereby lead reduction of imaging time in MR. To adopt the matching pursuit algorithm to the adaptive data acquisition in MRI, we have designed a dictionary which contains a windowed Fourier basis set. Because the basis set is localized spatially, the image signal could be divided into segmented signals so that matching pursuit with the segmented signals could lead to effective and object-dependent data acquisition. To verify the proposed technique, computer simulations and experiments are performed with a 1.0 T whole body MRI system.

Algorithms↗

Interaliasing distance of the flow convergence surface for determining mitral regurgitant volume: a validation study in a chronic animal model.

OBJECTIVES: We aimed to validate a new flow convergence (FC) method that eliminated the need to locate the regurgitant orifice and that could be performed semiautomatedly. BACKGROUND: Complex and time-consuming features of previously validated color Doppler methods for determining mitral regurgitant volume (MRV) have prevented their widespread clinical use. METHODS: Thirty-nine different hemodynamic conditions in 12 sheep with surgically created flail leaflets inducing chronic mitral regurgitation were studied with two-dimensional (2D) echocardiography. Color Doppler M-mode images along the centerline of the accelerating flow towards the mitral regurgitation orifice were obtained. The distance between the two first aliasing boundaries (interaliasing distance [IAD]) was measured and the FC radius was mathematically derived according to the continuity equation (R(calc) = IAD/(1 - radicalv(1)/v(2)), v(1) and v(2) being the aliasing velocities). The conventional 2D FC radius was also measured (R(meas)). Mitral regurgitant volume was then calculated according to the FC method using both R(calc) and R(meas). Aortic and mitral electromagnetic (EM) flow probes and meters were balanced against each other to determine the reference standard MRV. RESULTS: Mitral regurgitant volume calculated from R(calc) and R(meas) correlated well with EM-MRV (y = 0.83x + 5.17, r = 0.90 and y = 1.04x + 0.91, r = 0.91, respectively, p < 0.001 for both). However, both methods resulted in slight overestimation of EM-MRV (Delta was 3.3 +/- 2.1 ml for R(calc) and 1.3 +/- 2.3 ml for R(meas)). CONCLUSIONS: Good correlation was observed between MRV derived from R(calc) (IAD method) and EM-MRV, similar to that observed with R(meas) (conventional FC method) and EM-MRV. The R(calc) using the IAD method has an advantage over conventional R(meas) in that it does not require spatial localization of the regurgitant orifice and can be performed semiautomatedly.

Animals↗

Absolute concentrations of high-energy phosphate metabolites in normal, hypertrophied, and failing human myocardium measured noninvasively with (31)P-SLOOP magnetic resonance spectroscopy.

OBJECTIVE: The purpose of the present study was to measure absolute concentrations of phosphocreatine (PCr) and adenosine triphosphate (ATP) in normal, hypertrophied, and failing human heart. BACKGROUND: Conflicting evidence exists on the extent of changes of high-energy phosphate metabolites in hypertrophied and failing human heart. Previous reports using phosphorus-31 magnetic resonance spectroscopy ((31)P-MRS) have quantified metabolites in relative terms only. However, this analysis cannot detect simultaneous reductions. METHODS: Four groups of subjects (n = 10 each), were studied: volunteers and patients with hypertensive heart disease (HHD), aortic stenosis, and dilated cardiomyopathy (DCM). Left ventricular (LV) function and mass were measured by cine magnetic resonance imaging. Absolute and relative concentrations of PCr and ATP were determined by (31)P-MRS with spatial localization with optimum point spread function. RESULTS: Left ventricular ejection fraction remained normal in HHD and aortic stenosis, but was severely reduced to 18% in DCM; LV mass was increased by 55%, 79%, and 68% respectively. In volunteers, PCr and ATP concentrations were 8.82 +/- 1.30 mmol/kg wet weight and 5.69 +/- 1.02 mmol/kg wet weight, and the PCr/ATP ratio was 1.59 +/- 0.33. High-energy phosphate levels were unaltered in HHD. In aortic stenosis, PCr was decreased by 28%, whereas ATP remained constant. In DCM, PCr was reduced by 51%, ATP by 35%, and reduction of the PCr/ATP ratio by 25% was of borderline significance (p = 0.06). Significant correlations were observed among energetic and functional variables, with the closest relations for PCr. CONCLUSIONS: In human heart failure due to DCM, both PCr and ATP are significantly reduced. Ratios of PCr to ATP underestimate changes of high-energy phosphate levels.

Adenosine Triphosphate↗

Nuclear magnetic resonance imaging-guided phosphorus-31 spectroscopy of the human heart.

Phosphorus-31 nuclear magnetic resonance spectroscopy can determine the status of high energy phosphates in vivo. However, its application to human cardiac studies requires precise spatial localization without significant contamination from other tissues. Using image-selected in-vivo spectroscopy (ISIS), a technique that allows three-dimensional localization of the volume of interest, 12 subjects were studied to determine the feasibility and reproducibility of phosphorus-31 spectroscopy of the human heart. Nuclear magnetic resonance imaging was performed using a commercial 1.5 tesla system to define the volume of interest. Phosphorus-31 spectra were obtained from the septum and anteroapical region of the left ventricle in 10 studies. Relative peak heights and areas were determined for high energy phosphates. The mean phosphocreatine to adenosine triphosphate ratio was 1.33 +/- 0.19 by height analysis and 1.23 +/- 0.27 by area analysis. Duplicate measurements in four subjects showed a reproducibility of less than or equal to 10% in three of the subjects. All spectra showed significant signal contribution from the 2,3 diphosphoglycerate in chamber red cells without evidence of skeletal muscle contamination. These results demonstrate the feasibility of image-guided phosphorus-31 spectroscopy for human cardiac studies and indicate the potential of this technique to study metabolic disturbances in human myocardial disease.

Adenosine Triphosphate↗

A wavelet-based method for removal of out-of-plane structures in digital tomosynthesis.

Reconstructed images in digital tomosynthesis (DTS) are affected by artifacts due to blur from planes other than the fulcrum plane. A wavelet-based method has been developed for the discrimination and subsequent removal of unrelated structures from the reconstructed plane. The approach exploits both the specific pattern of noise in DTS and the spatial locality of the wavelet transformation. The technique was implemented on a DTS clinical protoype system. Experimental evaluation on angiographic types of images demonstrated excellent noise differentiation and elimination. The method is therefore particularly useful for certain medical imaging applications such as vascular DTS imaging.

Angiography↗

Zebrafish lunatic fringe demarcates segmental boundaries.

Cell interactions involving Notch signaling are required for the demarcation of tissue boundaries in both invertebrate and vertebrate development. Members of the Fringe gene family encode beta-1,3 N-acetyl-glucosaminyltransferases that function to refine the spatial localization of Notch-receptor signaling to tissue boundaries. In this paper we describe the isolation and characterization of the zebrafish (Danio rerio) homologue of the lunatic fringe gene (lfng). Zebrafish lfng is generally expressed in equivalent structures to those reported for the homologous chick and mouse genes. These sites include expression along the A-P axis of the neural tube, within the lateral plate mesoderm, in the presomitic mesoderm and the somites and in specific rhombomeres of the hindbrain; however, within these general expression domains species-specific differences in lfng expression exist. In mouse, Lfng is expressed in odd-numbered rhombomeres, whereas in zebrafish, expression occurs in even-numbered rhombomeres. In contrast to reports in both mouse and chicken embryos showing a kinematic cyclical expression of Lfng mRNA in the presomitic paraxial mesoderm, we find no evidence for a cyclic pattern of expression for the zebrafish lfng gene; instead, the zebrafish lfng is expressed in two static stripes within the presomitic mesoderm. Nevertheless, in zebrafish mutants affecting the correct formation of segment boundaries in the hindbrain and somites, lfng expression is aberrant or lost.

Amino Acid Sequence↗

Differential Pax6 promoter activity and transcript expression during forebrain development.

Three different Pax6 promoters -- P0, P1, and P alpha -- show differential activity in the developing eye and spinal cord. To examine promoter usage during forebrain development, we performed in situ hybridization and reverse transcription-polymerase chain reaction to detect transcripts initiated from each promoter. Promoter-specific transcripts are expressed within subdomains of total Pax6 expression, but differ from one another in their spatial localization and expression over time. Additionally, we identified a novel P0-initiated transcript and detected a developmentally regulated antisense transcript.

Alternative Splicing↗

A three-enzyme microelectrode sensor for detecting purine release from central nervous system.

As the purines, in particular adenosine, are important signaling agents in the nervous system we have devised a new biosensor for directly measuring their production in real time during physiological activity. Our amperometric adenosine biosensor is made by entrapping 3 enzymes (xanthine oxidase, purine nucleoside phosphorylase and adenosine deaminase) in a composite lactobionamide and amphiphillic polypyrrole matrix around a Pt microelectrode. The resulting sensors are small (25-100 microm diameter), fast responding (10-90% rise time, 2+/-0.23 s), sensitive (100-222 mA M(-1) cm(-2)) and stable (100% activity after 5 days). The sensor was used in vivo to demonstrate the spatial localization of release of adenosine from Xenopus embryo spinal cord during fictive swimming.

Animals↗

Axial patterning in leaves and other lateral organs.

The establishment of abaxial-adaxial polarity in lateral organs involves factors intrinsic to the primordia and interactions with the apical meristem from which they are derived. Recent molecular genetic studies have identified some of the genes that promote either adaxial or abaxial cell fates, with many of the genes encoding spatially localized transcription factors.

Animals↗

Phosphoinositides as spatial regulators of membrane traffic.

Inositol phospholipids are a focus of renewed interest with the discovery of their unanticipated pivotal roles in membrane trafficking events. Reversible phosphorylation of phosphatidylinositol generates spatially localized signals on membranes that recruit or activate proteins essential for cell membrane budding, fission and fusion. Recent advances have taken place in the characterization of lipid kinases and phosphoinositide-regulated effector proteins, and in the elucidation of phospholipase D mediated mechanisms involving ADP ribosylation factor and Rho family proteins. The roles played by phosphoinositides in aspects of secretory granule formation, fusion and endocytosis indicate the importance of phosphorylated lipids for neurotransmitter release.

Animals↗

Valence excitations in electron microscopy: resolved and unresolved issues.

Recent progress in the interpretation of spatially localized valence loss spectra is outlined. For a well-defined geometry of dielectric interfaces, detailed and quantitative analysis is now possible. In other cases, useful results may still be available although the common assumption that the spectrum at each point should be directly related to that from a uniform reference sample of appropriate composition may not always be valid.

Journal Article↗

The surgical approach to orbital space-occupying lesions.

From January 1985 to April 1987, 60 patients with orbital space-occupying lesions were treated surgically. Orientation and spatial localization of the lesion had already been achieved preoperatively by means of imaging methods. The decision on whether to use a neurosurgical transcranial, or a facial surgical approach, was made on this basis. The less elaborate procedure in each case was given preference to obtain a representative tissue sample. The definitive planning of the operation was then carried out with knowledge of the histology: a transcranial operation alone or as an additional measure was only necessary in six patients. Either definitive tumour therapy could be performed with or without temporal osteotomy in the remaining 54 patients exclusively via an anterior orbitotomy, or further oncological treatment was commenced after histological verification of the diagnosis.

Humans↗

Fine structure of the biogenesis of Giardia lamblia encystation secretory vesicles.

Synthesis, transport, and assembly of the extracellular cyst wall is the hallmark of Giardia lamblia encystation. Much is known of the biochemical pathways and their regulation. However, from a cell biology point of view, the biogenesis of the encystation specific vesicles (ESVs) that transport cyst wall proteins to the periphery of the cell is poorly understood. Therefore, we exploited a number of complementary ultrastructural approaches to test the hypothesis that the formation of ESVs utilizes a novel regulated secretory pathway. We analyzed parasites at different stages of encystation in vitro by electron microscopy of thin sections, freeze fracture replicas, and three-dimensional reconstruction from serial sections of cells fixed for cytochemical localization of the endoplasmic reticulum (ER) marker, glucose 6-phosphatase. We also used a stereological approach to determine the area occupied by the ER, clefts, ESVs, and cyst wall. Taken together, our kinetic data suggest that some ER cisternae first dilate to form clefts, which enlarge into the ESVs. Living non-encysting and early-encysting trophozoites were labeled around the periphery of both nuclei with C(6)-NBD-ceramide. At 18-21 h, outward migration of some ESVs frequently caused protrusions at the periphery of encysting trophozoites. The presence of lysosome-like peripheral vesicles between the ESV and plasma membrane of the cell was confirmed using acridine orange, an acidic compartment marker. Our data suggest that G. lamblia has a novel secretory pathway in which certain functions of the ER and Golgi co-localize spatially and temporally. These studies will increase understanding of the evolutionary appearance of regulated secretory pathways for assembly of a primitive extracellular matrix in an early diverging eukaryote.

Animals↗

Characterization of the first definitive hematopoietic stem cells in the AGM and liver of the mouse embryo.

At day 10 in mouse gestation, the intraembryonic aorta-gonads-mesonephros (AGM) region generates the first definitive hematopoietic stem cells (HSCs) of the adult blood system. By 11 days postcoitum, the liver contains such HSCs. While HSCs of the adult bone marrow and late-stage fetal liver have been extensively characterized for cell surface markers, there has been no phenotypic description of the first HSCs during embryo development. We report here the temporal cell surface phenotype of HSCs from the AGM region and early fetal liver and show that all HSCs reside in the c-kit+ population. c-kit+ HSCs from AGM and liver are mainly CD34+ and in the AGM are in both Mac-1+ and Mac-1 fractions. These results demonstrate that during mouse ontogeny the first definitive HSCs are similar in cell surface phenotype to the HSCs of adult bone marrow but that spatial localization and developmental time are critical factors in the phenotypic assessment of this functional cell population.

Animals↗

Radiation damage to DNA: the importance of track structure.

A wide variety of biological effects are induced by ionizing radiation, from cell death to mutations and carcinogenesis. The biological effectiveness is found to vary not only with the absorbed dose but also with the type of radiation and its energy, i.e., with the nature of radiation tracks. An overview is presented of some of the biological experiments using different qualities of radiation, which when compared with Monte Carlo track structure studies, have highlighted the importance of the localized spatial properties of stochastic energy deposition on the nanometer scale at or near DNA. The track structure leads to clustering of damage which may include DNA breaks, base damage etc., the complexity of the cluster and therefore its biological repairability varying with radiation type. The ability of individual tracks to produce clustered damage, and the subsequent biological response are important in the assessment of the risk associated with low-level human exposure. Recent experiments have also shown that biological response to radiation is not always restricted to the 'hit' cell but can sometimes be induced in 'un-hit' cells near by.

Animals↗