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At least 757 records · Page 42Linked to original sources
PubMed2001

Controlling tumor-derived and vascular endothelial cell growth: role of the 4Ff2 cell surface antigen.

We have isolated a monoclonal antibody, clone betaE11, which recognizes an antigen that is highly abundant on the surface of mitotic vascular endothelial cells and tumor cells. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, expression of this 190-kd antigen is approximately threefold higher in mitotic versus interphase endothelial cells. Treatment of tumor cells with an antibody to the betaE11 antigen inhibits their growth in a dose-dependent manner in vitro with maximal inhibition at an antibody concentration of 1 microg/ml. Different tumor cell lines demonstrate varying sensitivities to anti-betaE11 with the following order of growth inhibition: colon > prostate = glioma > melanoma = fibroblast > breast > liver. Furthermore, the betaE11 antigen localizes to regions of prostatic intraductal neoplasia in paraffin-embedded sections. Mass spectrometry of the cell-derived betaE11 protein and V8-protease fingerprint analysis indicate that the betaE11 antigen is nearly identical to the 4F2 heavy chain antigen, a cell surface protein that has been implicated in cell activation and proliferation. Expression of the betaE11 antigen during mitosis functionally links it to a fundamental aspect of cell proliferation, and its spatial localization on the surface of both proliferating endothelium and tumor cells demonstrates its potential for tumor immunotherapy.

Animals↗
PubMed2000

Local marangoni number at the onset of hydrothermal waves

We report the results of an experiment in which a layer of fluid, placed in a container open to the air, is subjected to a horizontal temperature difference DeltaT. The fluid height h is kept constant and both the horizontal temperature difference and the horizontal dimensions of the fluid layer are changed. In this configuration, when DeltaT goes beyond a certain threshold, waves propagating from the cold to the hot side appear (hydrothermal waves) with a determined group velocity. We study the influence of the container geometry on these waves. Close to the threshold, they are spatially localized near the hot side and a local Marangoni number is defined to describe this behavior. If DeltaT is further increased, the waves fill the whole fluid layer. We also find an agreement between our experimental results and theoretical works and simulations.

Journal Article↗
PubMed1999

Random dot motion stimuli are more sensitive than light stimuli for detection of visual field loss in ocular hypertension patients.

PURPOSE: To determine whether motion detection perimetry or luminance size threshold perimetry (a test using the motion perimetry method with luminance stimuli) is more sensitive in detecting visual loss in ocular hypertension patients. METHODS: Motion perimetry uses a customized computer graphics program to detect a subject's ability to identify a coherent shift in position of moving dots in a defined circular area against a background of fixed dots. Motion size threshold is defined as the smallest circular area within which dot motion is detected. Patients respond by touching the area of the computer monitor where they perceive the stimulus with a light pen. The localization errors are measured as the number of pixels from target center for each trial. Luminance size threshold perimetry uses the same technique except the background is dark gray and the stimuli are filled lighter gray circles. We tested one eye in each of 27 ocular hypertension patients and 27 age-matched normal subjects with both tests. Our main outcome measures were motion and luminance size thresholds, total deviation probability plot data, and spatial localization errors. RESULTS: With the total deviation probability plot analysis, the ocular hypertension patients had a greater number of abnormal test locations with motion perimetry stimuli than with luminance stimuli. The abnormal test points were located most often in the superior and inferior nasal regions. Six subjects had nerve fiber bundle-like defects to motion stimuli whereas three patients had defects with luminance size threshold perimetry. The ocular hypertension patients had significantly greater localization errors than the controls with both tests. CONCLUSIONS: Using a size thresholding technique in ocular hypertension patients, random dot motion stimuli appear to be more sensitive than luminance stimuli. Errors in stimulus localization are significantly increased in ocular hypertension patients, independent of the stimulus (motion or luminance) used.

Adult↗
PubMed2005

Incremental online learning in high dimensions.

Locally weighted projection regression (LWPR) is a new algorithm for incremental nonlinear function approximation in high-dimensional spaces with redundant and irrelevant input dimensions. At its core, it employs nonparametric regression with locally linear models. In order to stay computationally efficient and numerically robust, each local model performs the regression analysis with a small number of univariate regressions in selected directions in input space in the spirit of partial least squares regression. We discuss when and how local learning techniques can successfully work in high-dimensional spaces and review the various techniques for local dimensionality reduction before finally deriving the LWPR algorithm. The properties of LWPR are that it (1) learns rapidly with second-order learning methods based on incremental training, (2) uses statistically sound stochastic leave-one-out cross validation for learning without the need to memorize training data, (3) adjusts its weighting kernels based on only local information in order to minimize the danger of negative interference of incremental learning, (4) has a computational complexity that is linear in the number of inputs, and (5) can deal with a large number of-possibly redundant-inputs, as shown in various empirical evaluations with up to 90 dimensional data sets. For a probabilistic interpretation, predictive variance and confidence intervals are derived. To our knowledge, LWPR is the first truly incremental spatially localized learning method that can successfully and efficiently operate in very high-dimensional spaces.

Algorithms↗
PubMed2000

New insights into the hemodynamic blood oxygenation level-dependent response through combination of functional magnetic resonance imaging and optical recording in gerbil barrel cortex.

Fast, low-angle shoot functional magnetic resonance imaging (fMRI), based on the blood oxygenation level-dependent (BOLD) effect, was combined with optical recording of intrinsic signals (ORIS) and 2-deoxyglucose labeling in gerbil barrel cortex. We observed over the activated barrel a positive BOLD signal and increased levels of deoxyhemoglobin and total hemoglobin during each period of prolonged (30 sec) D2 vibrissal stimulation. These data show that the hemodynamic basis of this fMRI signal is not necessarily a washout of deoxyhemoglobin, as generally assumed. Instead, they suggest that a positive BOLD signal can also be caused by a local increase of blood volume, even if deoxyhemoglobin levels are persistently elevated. We also show that this alternative interpretation is consistent with theoretical models of the BOLD signal. The changes in BOLD signal and blood volume, which are most tightly correlated with the periodic stimulation, peak at the site of neuronal activation. These results contribute to the understanding of the hemodynamic mechanisms underlying the BOLD signal and also suggest analysis methods, which improve the spatial localization of neuronal activation with both fMRI and ORIS.

Animals↗
PubMed1998

Statistical analysis of functional MRI data in the wavelet domain.

The use of the wavelet transform is explored for the detection of differences between brain functional magnetic resonance images (fMRI's) acquired under two different experimental conditions. The method benefits from the fact that a smooth and spatially localized signal can be represented by a small set of localized wavelet coefficients, while the power of white noise is uniformly spread throughout the wavelet space. Hence, a statistical procedure is developed that uses the imposed decomposition orthogonality to locate wavelet-space partitions with large signal-to-noise ratio (SNR), and subsequently restricts the testing for significant wavelet coefficients to these partitions. This results in a higher SNR and a smaller number of statistical tests, yielding a lower detection threshold compared to spatial-domain testing and, thus, a higher detection sensitivity without increasing type I errors. The multiresolution approach of the wavelet method is particularly suited to applications where the signal bandwidth and/or the characteristics of an imaging modality cannot be well specified. The proposed method was applied to compare two different fMRI acquisition modalities. Differences of the respective useful signal bandwidths could be clearly demonstrated; the estimated signal, due to the smoothness of the wavelet representation, yielded more compact regions of neuroactivity than standard spatial-domain testing.

Adult↗
PubMed2002

Activation mechanisms of protein kinase C: maturation, catalytic activation, and targeting.

The biological function of protein kinase C (PKC) depends on its catalytic activity and spatial localization. Its catalytic competence and localization in the resting state are regulated by serine/threonine phosphorylations, i.e., "maturation." Upon stimulation of various receptors, PKC is catalytically activated by several activators including diacylglycerol. In addition, PKC often translocates to particular subcellular compartments including the plasma membrane and Golgi complex, and such translation is here referred to as "targeting." In short, the physiological function of PKC is controlled by the three events: maturation, catalytic activation, and targeting. Catalytic activation and targeting contribute to temporal, spatial, and isotype-specific regulation of PKC. This review summarizes the evidence for the role of these three events in the isotype-specific activation of PKC, with particular emphasis on catalytic activation and targeting by lipid mediators.

Calcium↗
PubMed1998

A localized double-quantum filter for the in vivo detection of brain glucose.

A double-quantum filter (DQF) sequence with PRESS localization was developed for in vivo detection of the glucose resonances in the 3.85-ppm region of the brain proton spectrum. The efficiency and spectral editing characteristics were studied in phantom and animal experiments. Approximately 45% detection efficiency was achieved at 4.7 T with TE = 68 ms. Since the efficiency of the DQF method is dependent on the relative phases of the RF pulses, a phase calibration procedure was used to correct for phase shifts induced by the spatial localization. In addition to detecting the 3.85-ppm glucose resonances with approximately 45% efficiency, the DQF sequence simultaneously detects 1.3-ppm lactate resonance with approximately 20% efficiency. The use of the DQF technique for simultaneously monitoring both the input and output of anaerobic glycolysis in the brain was demonstrated by detecting brain glucose and lactate in the same acquisition after iv injection of glucose followed by the induction of global ischemia.

Animals↗
PubMed2005

Global competition dictates local suppression in pattern rivalry.

Certain pairs of visual patterns, when superimposed as if transparent, elicit a wavering percept in which one or the other pattern can enjoy temporary periods of exclusive visibility. This multistable perceptual phenomenon is called monocular rivalry or pattern rivalry (PR) and is most pronounced when the component patterns are faint and devoid of detail. The principal mechanisms that give rise to PR continue to be a topic of speculation. In the present study, we examine the determinants of exclusive dominance during PR using a novel stimulus in which a central portion is free of conflict. By observing the properties of suppression in this so-called rivalry-free region, we demonstrate that perception is driven largely by the global and holistic interpretation of the patterns, rather than by the need to resolve local spatial conflict. The suppression of this central region was often complete and varied as a function of the parameters of the global stimulus, including the size of the surround region, its ocular configuration, and stereoscopic depth ordering. Suppression also varied as a function of pattern continuity across the central region as well as with the temporal offset of the overlapping components. These findings demonstrate that the visibility or invisibility of a pattern is not fundamentally a product of local processing, but is instead shaped by the brain's global interpretive assumptions regarding the composition of the stimulus.

Adolescent↗
PubMed2005

FRET analyses of the U2AF complex localize the U2AF35/U2AF65 interaction in vivo and reveal a novel self-interaction of U2AF35.

We have analyzed the interaction between the U2AF subunits U2AF35 and U2AF65 in vivo using fluorescence resonance energy transfer (FRET) microscopy. U2 snRNP Auxiliary Factor (U2AF) is an essential pre-mRNA splicing factor complex, comprising 35-kDa (U2AF35) and 65-kDa (U2AF65) subunits. U2AF65 interacts directly with the polypyrimidine tract and promotes binding of U2 snRNP to the pre-mRNA branchpoint, while U2AF35 associates with the conserved AG dinucleotide at the 3' end of the intron and has multiple functions in the splicing process. Using two different approaches for measuring FRET, we have identified and spatially localized sites of direct interaction between U2AF35 and U2AF65 in vivo in live cell nuclei. While U2AF is thought to function as a heterodimeric complex, the FRET data have also revealed a novel U2AF35 self-interaction in vivo, which is confirmed in vitro using biochemical assays. These results suggest that the stoichiometry of the U2AF complex may, at least in part, differ in vivo from the expected heterodimeric complex. The data show that FRET studies offer a valuable approach for probing interactions between pre-mRNA splicing factors in vivo.

Bacterial Proteins↗
PubMed2002

Hyphal tip-associated localization of Cdc42 is F-actin dependent in Candida albicans.

The rho-type GTPase Cdc42 is important for the establishment and maintenance of eukaryotic cell polarity. To examine whether Cdc42 is regulated during the yeast-to-hypha transition in Candida albicans, we constructed a green fluorescence protein (GFP)-Cdc42 fusion under the ACT1 promoter and observed its localization in live C. albicans cells. As in Saccharomyces cerevisiae, GFP-Cdc42 was observed around the entire periphery of the cell. In yeast-form cells of C. albicans, it clustered to the tips and sides of small buds as well as to the mother-daughter neck region of large-budded cells. Upon hyphal induction, GFP-Cdc42 clustered to the site of hyphal evagination and remained at the tips of the hyphae. This temporal and spatial localization of Cdc42 suggests that its activity is regulated during the yeast-to-hypha transition. In addition to the accumulation at the hyphal tip, GFP-Cdc42 was also seen as a band within the hyphal tube in cells that had undergone nuclear separation. With the F-actin-assembly inhibitor latrunculin A, we found that GFP-Cdc42 accumulation at the bud site in yeast-form cells is F-actin independent, whereas GFP-Cdc42 accumulation at the hyphal tip requires F-actin. Furthermore, disruption of the F-actin cytoskeleton impaired the transcriptional induction of hypha-specific genes. Therefore, hypha formation resembles mating in Saccharomyces cerevisiae in that both require F-actin for GFP-Cdc42 localization and efficient signaling.

Actins↗
PubMedDate not supplied

Delineation and quantitation of brain lesions by fuzzy clustering in positron emission tomography.

In this study, we investigate the application of the fuzzy clustering to the anatomical localization and quantitation of brain lesions in Positron Emission Tomography (PET) images. The method is based on the Fuzzy C-Means (FCM) algorithm. The algorithm segments the PET image data points into a given number of clusters. Each cluster is an homogeneous region of the brain (e.g. tumor). A feature vector is assigned to a cluster which has the highest membership degree. Having the label affected by the FCM algorithm to a cluster, one may easily compute the corresponding spatial localization, area and perimeter. Studies concerning the evolution of a tumor after different treatments in two patients are presented.

Algorithms↗
PubMed2003

Colocalization of ATP release sites and ecto-ATPase activity at the extracellular surface of human astrocytes.

Extracellular ATP and other nucleotides function as autocrine and paracrine signaling factors in many tissues. Recent studies suggest that P2 nucleotide receptors and ecto-nucleotidases compete for a limited pool of endogenously released nucleotides within cell surface microenvironments that are functionally segregated from the bulk extracellular compartment. To test this hypothesis, we have used luciferase-based methods to continuously record extracellular ATP levels in monolayers of human 1321N1 astrocytoma cells under resting conditions, during stimulation of Ca2+-mobilizing receptors for thrombin or acetylcholine, and during mechanical stimulation by hypotonic stress. Soluble luciferase was utilized as an indicator of ATP levels within the bulk extracellular compartment, whereas a chimeric protein A-luciferase, adsorbed to antibodies against a glycosylphosphatidylinositol-anchored plasma membrane protein, was used as a spatially localized probe of ATP levels at the immediate extracellular surface. Significant accumulation of ATP in the bulk extracellular compartment, under either resting (1-2 nm ATP) or stimulated (10-80 nm ATP) conditions, was observed only when endogenous ecto-ATPase activity was pharmacologically inhibited by the poorly metabolizable analog, betagamma-methylene ATP. In contrast, accumulation of submicromolar ATP in the cell surface microenvironment was readily measured even in the absence of ecto-ATPase inhibition suggesting that the spatially colocalized luciferase could effectively compete with endogenous ecto-ATPases for released ATP. Other experiments revealed a critical role for elevated cytosolic [Ca2+] in the ATP release mechanism triggered by thrombin or muscarinic receptors but not in basal ATP release or release stimulated by hypotonic stress. These observations suggest that ATP release sites are colocalized with ecto-ATPases at the astrocyte cell surface. This colocalization may act to spatially restrict the actions of released ATP as a paracrine or autocrine mediator of cell-to-cell signaling.

Adenosine Triphosphatases↗
PubMed2006

Feedback control of subcritical oscillatory instabilities.

Feedback control of a subcritical oscillatory instability is investigated in the framework of a globally-controlled complex Ginzburg-Landau equation that describes the nonlinear dynamics near the instability threshold. The control is based on a feedback loop between the system linear growth rate and the maximum of the amplitude of the emerging pattern. It is shown that such control can suppress the blow up and result in the formation of spatially localized pulses similar to oscillons. In the one-dimensional case, depending on the values of the linear and nonlinear dispersion coefficients, several types of the pulse dynamics are possible in which the computational domain contains: (i) a single stationary pulse; (ii) several coexisting stationary pulses; (iii) competing pulses that appear one after another at random locations so that at each moment of time there is only one pulse in the domain; (iv) spatiotemporally chaotic system of short pulses; (v) spatially-synchronized pulses. Similar dynamic behavior is found also in the two-dimensional case. The effect of the feedback delay is also studied. It is shown that the increase of the delay leads to an oscillatory instability of the pulses and the formation of pulses with oscillating amplitude.

Journal Article↗
PubMed1995

Calmodulin binding to Drosophila NinaC required for termination of phototransduction.

The ninaC locus encodes two unconventional myosins, p132 and p174, consisting of fused protein kinase and myosin head domains expressed in Drosophila photoreceptor cells. NinaC are the major calmodulin-binding proteins in the retina and the NinaC-calmodulin interaction is required for the normal subcellular localization of calmodulin as well as for normal photo-transduction. In the current report, we present evidence for two calmodulin-binding sites in NinaC, C1 and C2, which have different in vitro binding properties. C1 was found to be common to both p132 and p174 while C2 was unique to p174. To address the requirements for calmodulin binding at each site in vivo, we generated transgenic flies expressing ninaC genes deleted for either C1 or C2. We found that the spatial localization of calmodulin depended on binding to both C1 and C2. Furthermore, mutation of either site resulted in a defective photoresponse. A prolonged depolarization afterpotential (PDA) was elicited at lower light intensities than necessary to produce a PDA in wild-type flies. These results suggest that calmodulin binding to both C1 and C2 is required in vivo for termination of phototransduction.

Amino Acid Sequence↗
PubMed1997

Golgi membrane skeleton: identification, localization and oligomerization of a 195 kDa ankyrin isoform associated with the Golgi complex.

To extend our finding of a Golgi-localized form of the membrane skeleton protein spectrin, we have identified an isoform of ankyrin that associates at steady state with the Golgi complex. Immuno-light and -electron microscopy show that this ankyrin isoform localizes to the perinuclear cytoplasm on tubular vesicular structures that co-stain with Golgi marker proteins. An antiserum raised against erythrocyte ankyrin, which was used to identify the Golgi ankyrin, recognized three prominent polypeptides of 220, 213 and 195 kDa in MDCK cells. Affinity purification of this antiserum against each of these MDCK cell ankyrins revealed that only an antibody specific for the 195 kDa form retained the ability to stain the Golgi complex; affinity purified antibody preparations specific for both the 220 and 213 kDa forms stained punctate and reticular cytoplasmic structures distinct from the Golgi complex. Antibody specific for the 195 kDa ankyrin did not recognize a recently identified 119 kDa ankyrin that is also localized to the Golgi. The 195 kDa Golgi ankyrin binds purified erythrocyte spectrin, and rapidly co-sediments with Golgi beta-spectrin during brief, low speed centrifugation of Triton X-100 extracts of MDCK cells. Golgi ankyrin and beta-spectrin are retained on tubular vesicular 'Golgi ghosts' following extraction of cultured cells with Triton X-100. Significantly, Golgi ghost tubules containing ankyrin/spectrin are co-linear with individual microtubules, suggesting a role for both Golgi membrane skeleton and microtubules in spatial localization of the Golgi. Golgi ankyrin dissociates from Golgi membranes during mitosis and in cells treated with brefeldin A, indicating that Golgi ankyrin has a dynamic assembly state similar to that of Golgi spectrin and other Golgi membrane coat proteins.

Animals↗
PubMed1991

13C NMR for the assessment of human brain glucose metabolism in vivo.

Proton-decoupled 13C NMR spectra of the human head were obtained during hyperglycemic glucose clamping using intravenous infusions of [1-13C]glucose in normal volunteers. In addition to 13C signals of mobile lipids, a variety of new metabolite resonances could be resolved for the first time in the human brain. At an enrichment level of 20% [1-13C]glucose, the signals of alpha- and beta-glucose at 92.7 and 96.6 ppm, respectively, could be detected in the human brain after only an infusion period of 15 min. The spatial localization of the different regions of interest was confirmed by 13C NMR spectroscopic imaging with a time resolution of 9 min. Increasing the enrichment level to 99% [1-13C]glucose not only improved the time resolution but allowed the detection of metabolic breakdown products of [1-13C]glucose. The time course of 13C label incorporation into the C2, C3, and C4 resonances of glutamate/glutamine and into lactate could be recorded in the human brain. These results suggest the possibility of obtaining time-resolved, spatially selective, and chemically specific information on the human body.

Adult↗
PubMed1993

MR imaging of fat-containing tissues: valuation of two quantitative imaging techniques in comparison with localized proton spectroscopy.

Since lipid protons, consisting mainly of triacylglycerols (TAG), are rather mobile, magnetic resonance imaging (MRI) is ideally suited for the examination of fat-containing tissues such as bone marrow. In contrast to water protons, however, lipid protons are chemically distinct and give rise to at least eight resonance peaks with different T1 and T2 relaxation times in the 1H spectrum. This is why the characterization of fat-containing tissues by quantitative MRI is much more difficult than that of most other tissues. In our study we wanted to examine the accuracy and the potential of a 1H chemical shift imaging (CSI) technique and a multiple spin-echo imaging (MSEI) technique. A stimulated-echo (STEAM) sequence for spatially localized proton spectroscopy was used as the reference method. In the first part of this paper, we describe quantitative imaging experiments which were performed to assess the accuracy of the fat-water separation according to the Dixon method and the bi-exponential decomposition of the MSEI data. For that purpose, we used a two-compartment phantom filled with either an aqueous Gd-DTPA solution and vegetable oil or with two different aqueous Gd-DTPA solutions, respectively. The analysis of the 1H CSI data revealed that the presence of non-methylen protons in neutral fats leads to a slight under-estimation (of about 15%) of the relative fat fraction. The error is described theoretically and verified quantitatively by STEAM measurements. The bi-exponential analysis of the transverse relaxation data, on the other hand, yields reliable T2 values if the relative proton density of both components is higher than 15%. IN the second part of our investigation, the same techniques were applied to acquire data from the subcutaneous fatty tissue, the femoral head, and the lumbar vertebrae of three healthy volunteers. In the bone marrow spectra, only two broad resonances could be resolved; they were superpositions of diverse molecular groups with different T1 and T2 relaxation times. In these cases, localized proton spectroscopy does not provide additional information with respect to 1H CSI. The MSEI data of the three examined fat containing tissue regions were adequately fitted by a bi-exponential function despite the fact that there were much more chemically distinct protons present in fatty tissues.

Adipose Tissue↗