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New approach to estimating variability in visual field data using an image processing technique.

AIMS: A new framework for evaluating pointwise sensitivity variation in computerised visual field data is demonstrated. METHODS: A measure of local spatial variability (LSV) is generated using an image processing technique. Fifty five eyes from a sample of normal and glaucomatous subjects, examined on the Humphrey field analyser (HFA), were used to illustrate the method. RESULTS: Significant correlation between LSV and conventional estimates--namely, HFA pattern standard deviation and short term fluctuation, were found. CONCLUSION: LSV is not dependent on normals' reference data or repeated threshold determinations, thus potentially reducing test time. Also, the illustrated pointwise maps of LSV could provide a method for identifying areas of fluctuation commonly found in early glaucomatous field loss.

Adult↗

Oscillatory convection in binary mixtures: thermodiffusion, solutal buoyancy, and advection.

The role of thermodiffusive generation of concentration fluctuations via the Soret effect, their contribution to the buoyancy forces that drive convection, the advective mixing effect of the latter, and the diffusive homogenisation are compared and elucidated for oscillatory convection. Numerically obtained solutions of the field equations in the form of spatially extended relaxed traveling waves, of standing waves, and of the transient growth of standing waves and their transition to traveling waves are discussed as well as spatially localized convective states of traveling waves that are surrounded by the quiescent fluid.

Journal Article↗

Dynamics of antagonistic neural processing elements.

Two coupled nonlinear first-order systems whose dynamic behavior reflects the neural states exhibited by a spatially localized population of excitatory and inhibitory nerve cells are described. The dynamics of each constituent neural subpopulation represents a fundamental neural information processing element (PE) of a complex neural system. Phase plane analysis is used in this paper to show how such antagonistic positive acting (excitatory) and negative acting (inhibitory) PEs can generate diverse steady-state and temporal phenomena when the nonlinear system parameters of the PEs are altered. By modifying a selected set of parameters, it is possible to program the positive and negative PEs to exhibit various dynamic attributes such as multiple stable states, transient response behavior and limit-cycle oscillations. These dynamic attributes may be used to perform a variety of useful computational tasks in signal processing and vision systems such as short-term memory (STM), temporal filtering (TF) and pulse frequency modulation (PFM). Computer simulations are presented throughout this paper in order to illustrate these dynamic attributes.

Algorithms↗

Polarized cell growth in higher plants.

Pollen tubes and root hairs are highly elongated, cylindrically shaped cells whose polarized growth permits them to explore the environment for the benefit of the entire plant. Root hairs create an enormous surface area for the uptake of water and nutrients, whereas pollen tubes deliver the sperm cells to the ovule for fertilization. These cells grow exclusively at the apex and at prodigious rates (in excess of 200 nm/s for pollen tubes). Underlying this rapid growth are polarized ion gradients and fluxes, turnover of cytoskeletal elements (actin microfilaments), and exocytosis and endocytosis of membrane vesicles. Intracellular gradients of calcium and protons are spatially localized at the growing apex; inward fluxes of these ions are apically directed. These gradients and fluxes oscillate with the same frequency as the oscillations in growth rate but not with the same phase. Actin microfilaments, which together with myosin generate reverse fountain streaming, undergo rapid turnover in the apical domain, possibly being regulated by key actin-binding proteins, e.g., profilin, villin, and ADF/cofilin, in concert with the ion gradients. Exocytosis of vesicles at the apex, also dependent on the ion gradients, provides precursor material for the continuously expanding cell wall of the growing cell. Elucidation of the interactions and of the dynamics of these different components is providing unique insight into the mechanisms of polarized growth.

Actin Cytoskeleton↗

Attentional modulation of visual processing.

Single-unit recording studies in the macaque have carefully documented the modulatory effects of attention on the response properties of visual cortical neurons. Attention produces qualitatively different effects on firing rate, depending on whether a stimulus appears alone or accompanied by distracters. Studies of contrast gain control in anesthetized mammals have found parallel patterns of results when the luminance contrast of a stimulus increases. This finding suggests that attention has co-opted the circuits that mediate contrast gain control and that it operates by increasing the effective contrast of the attended stimulus. Consistent with this idea, microstimulation of the frontal eye fields, one of several areas that control the allocation of spatial attention, induces spatially local increases in sensitivity both at the behavioral level and among neurons in area V4, where endogenously generated attention increases contrast sensitivity. Studies in the slice have begun to explain how modulatory signals might cause such increases in sensitivity.

Action Potentials↗

In vivo MR spectroscopic imaging with P-31. Work in progress.

Magnetic resonance imaging techniques were applied to in vivo spectroscopic analysis of spatially resolved phosphorus spectra in the cat brain to determine whether changes associated with stroke could be detected. Two-dimensional images of separate phosphorus-containing compounds, as well as spectra arising from spatially localized points, demonstrated that metabolism within tissue could be monitored in this manner. Preliminary results of phosphorus imaging of the human body in vivo are reported.

Animals↗

Chronic adult cerebral infarction studied by phosphorus NMR spectroscopy.

Spatially localized phosphorus (P-31) nuclear magnetic resonance (NMR) spectra were recorded in vivo from the brains of nine healthy adults and of four adult patients examined 10 weeks to 7 years following cerebral infarction. Spectra from the healthy adult subjects showed reduced inorganic phosphate (Pi) and phosphoester resonances compared with earlier such measurements in infants. Spectra from subjects with chronic cerebral infarction exhibited no significant differences in the ratios of high-energy phosphate metabolites, Pi, or pH compared with the spectra of the healthy adults, although decreases of up to about 40% in the total P-31 NMR signal from infarcted tissue were noted. These results are consistent with a reduction in the number of metabolically active brain cells and/or a decrease in cellular functional activity in chronic infarction. Phosphorus metabolite ratios do not appear to be a sensitive indicator of abnormal energy reserve in chronic brain infarction in adults.

Adult↗

Human in vivo NMR spectroscopy in diagnostic medicine: clinical tool or research probe?

In this critical review of human in vivo nuclear magnetic resonance (NMR) spectroscopy, the questions of which chemical species can be detected and with what sensitivity, their biochemical significance, and their potential clinical value are addressed. The current in vivo detectability limit is about 10(-6) of that of tissue water protons, necessitating a 1-10 cm3-volume of tissue and approximately 10-minute averaging time. This permits access to fats, membrane lipid metabolism, high-energy phosphate metabolism, glycogen, some neurotransmitters and metabolites in the citric acid cycle, and artificially introduced fluorocompounds. While hydrogen-31, phosphorus-31, carbon-13, sodium-23, and fluorine-19 in vivo results are discussed, the majority of patient studies use P-31 NMR spectroscopy. Here results from metabolic and ischemic disorders substantiate a case for spectroscopy as a diagnostic modality. The use of a broad range of spatial localization strategies is justifiable on the basis of the location and size of the pathologic condition and of NMR sensitivity. Abnormalities in spectra from many other disorders, most notably cancer, and improvements are often observed early in the course of successful therapy. Yet the potential impact of these results on clinical diagnosis and therapeutic monitoring is not always well understood, and many questions remain. Neurotransmitters and citric acid cycle metabolites exhibit high H-1 NMR sensitivities and represent major untapped potential for human clinical spectroscopy research. Studies evaluating spectroscopy in the context of existing modalities are needed. The unique ability of spectroscopy to provide noninvasive information about tissue chemistry in patients bodes well for its impact on clinical research and disease diagnosis.

Amino Acids↗

Alzheimer dementia: quantification of energy metabolism and mobile phosphoesters with P-31 NMR spectroscopy.

To determine whether high-energy phosphate metabolism and mobile phosphoester indexes of membrane metabolism are altered in Alzheimer disease and to help resolve some inconsistencies in the literature, brain phosphate metabolite concentrations and ratios were measured in 11 patients with mild to severe dementia of the probable Alzheimer type and 14 healthy subjects. Fully relaxed, spatially localized, phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy and proton (hydrogen-1) MR imaging were performed. No significant differences were found in the concentrations and relative ratios of phosphocreatine, nucleoside triphosphate, inorganic phosphate, phosphomonoester, and phosphodiesters in whole axial sections through the lateral cerebral ventricles of the brain that could not be accounted for by atrophy. There was no correlation between P-31 NMR indexes and the severity of dementia as assessed with neuropsychologic testing. High-energy phosphate and membrane metabolism, as detected in vivo with P-31 NMR spectroscopy in whole-brain sections, do not appear to play a major role in the disease process, except as a direct consequence of atrophy quantified with H-1 MR imaging.

Aged↗

Transport properties of the lens.

Many studies have shown that the lens is a multicellular syncytial tissue whose electrophysiological properties are the integrated result of membrane transport, low-resistance gap junctions interconnecting the cells, and the restricted extracellular space between cells. There are at least three structurally distinct populations of cells within the lens, and the membrane transport properties of each cell type appear to differ. Indeed, there may be subcellular specialization of membrane transport properties in the surface epithelial cells. We review the physical structure of the lens, its electrical structure, and our present knowledge of the membrane transport properties of the different cell types. Our recent work has focused on radially circulating fluxes generated by the spatial localization of membrane transport in surface cell membranes versus inner fiber cell membranes. We review this work and present some simplified models of the results with some discussion of physiological implications.

Animals↗

Immunocytochemical and functional characterization of Na+ conductance in adult alveolar pneumocytes.

The purpose of this study was to document the existence, assess the spatial localization, and characterize some of the transport properties of proteins antigenically related to epithelial Na+ channels in freshly isolated rabbit and rat alveolar type II (ATII) cells. ATII cells, isolated by elastase digestion of lung tissue and purified by density-gradient centrifugation, were incubated with polyclonal antibodies raised against Na+ channel protein purified from beef kidney papilla (NaAb), followed by a secondary antibody (goat antirabbit immunoglobulin G conjugated to fluorescein isothiocyanate). Rat ATII cells exhibited specific staining with NaAb at the level of the plasma membrane, which, in most cells, colocalized with that of the lectin Maclura pomiferra agglutinin, an apical surface marker. In Western blots, NaAb specifically recognized a 135 +/- 10-kDa protein in rat ATII membrane vesicles. When patch clamped in the whole cell mode using symmetrical solutions (150 mM Na+ glutamate), ATII cells exhibited outwardly rectified Na+ currents that were diminished by amiloride (10-100 microM) instilled into the bath solution. Ion substitution studies showed that the conductive pathways were three times more permeable to Na+ than K+. Amiloride, benzamil, and 5-(N-ethyl-N-isopropyl)-2',4'-amiloride were equally effective in diminishing 22Na+ flux into rabbit and rat ATII cells (45% inhibition at 100 microM, with IC50 of approximately 1 microM for all inhibitors). Tetraethylammonium chloride (10 mM) or BaCl2 (2 mM), well-known K+ channel blockers, had no effect on 22Na+ uptake. These results indicate that ATII cells express an amiloride-sensitive Na+ conductance, probably a channel, with a lower affinity for amiloride and its structural analogues than the well-established amiloride-sensitive Na+ channels found in bovine renal papila and cultured amphibian A6 kidney cells.

Amiloride↗

G protein-coupled receptors in gastrointestinal physiology. III. Asymmetry in plasma membrane signal transduction: lessons from brush-border Na+/H+ exchangers.

Signal transduction in epithelial cells adds another level of complexity to the signaling that occurs in symmetrical cells, in the form of the need to coordinate and keep separate signals at the apical and basolateral membranes. Regulation by protein kinases of ileal NaCl absorption and its component brush-border Na+/H+ exchanger are used as an example of how signaling in epithelial cells must deal with spatial localization of signals, protein-protein interactions, signaling molecules, and the involvement of the transport protein being regulated in collecting and focusing the signals generated at the receptor and beyond.

Animals↗

Exercise training attenuates coronary smooth muscle phenotypic modulation and nuclear Ca2+ signaling.

Physical inactivity is an independent risk factor for coronary heart disease, yet the mechanism(s) of exercise-related cardioprotection remains unknown. We tested the hypothesis that coronary smooth muscle after exercise training would have decreased mitogen-induced phenotypic modulation and enhanced regulation of nuclear Ca(2+). Yucatan swine were endurance exercise trained (EX) on a treadmill for 16-20 wk. EX reduced endothelin-1-induced DNA content by 40% compared with sedentary (SED) swine (P < 0.01). EX decreased single cell peak endothelin-1-induced cytosolic Ca(2+) responses compared with SED by 16% and peak nuclear Ca(2+) responses by 33% (P < 0.05), as determined by confocal microscopy. On the basis of these results, we hypothesized that sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) and intracellular Ca(2+) stores in native smooth muscle are spatially localized to dissociate cytosolic Ca(2+) and nuclear Ca(2+). Subcellular localization of SERCA in living and fixed cells revealed a distribution of SERCA near the sarcolemma and on the nuclear envelope. These results show that EX enhances nuclear Ca(2+) regulation, possibly via SERCA, which may be one mechanism by which coronary smooth muscle cells from EX are less responsive to mitogen-induced phenotypic modulation.

Animals↗

P2X1 receptors mediate sympathetic postjunctional Ca2+ transients in mesenteric small arteries.

Brief, spatially localized Ca(2+) transients occur in the smooth muscle adjacent to perivascular nerves of small arteries during neurogenic contractions. We named these "junctional Ca(2+) transients" (jCaTs) and postulated that they arose from Ca(2+) entering smooth muscle cells through P2X(1) receptors activated by neurally released ATP. Nevertheless, the lack of potent, subtype-selective P2X-receptor antagonists made determining the exact molecular identity of the channels difficult. Here we used small, pressurized mesenteric arteries from P2X(1)-receptor-deficient mice (KO) to test the hypothesis that jCaTs arise from Ca(2+) entering the smooth muscle cell via P2X(1) receptors. In wild-type (WT) arteries, confocal microscopy of fluo-4 fluorescence during electrical field stimulation (EFS) of perivascular sympathetic nerves revealed jCaTs in the smooth muscle cells adjacent to the perivascular nerves, similar to those reported previously in rat arteries, and alpha-latrotoxin (2.5 nM) markedly increased the frequency of "spontaneous" jCaTs. In the KO arteries, however, neither EFS nor alpha-latrotoxin elicited any jCaTs. A potent P2X-receptor agonist, alpha,beta-methylene ATP (10.0 microM), elicited strong contractions and increased intracellular Ca(2+) concentration in WT arteries but elicited neither in KO arteries. A biphasic vasoconstriction in response to EFS was observed in WT arteries. In KO arteries, however, the initial rapid, transient component of the biphasic vasoconstriction was absent. The data support the hypothesis that jCaTs represent Ca(2+) that enters the smooth muscle cells through P2X(1) receptors activated by neurally released ATP and that this Ca(2+) is involved in the initial rapid component of the sympathetic neurogenic contraction.

Adenosine Triphosphate↗

Regulation of murine myocardial energy metabolism during adrenergic stress studied by in vivo 31P NMR spectroscopy.

Image-guided, spatially localized 31P magnetic resonance spectroscopy (MRS) was used to study in vivo murine cardiac metabolism under resting and dobutamine-induced stress conditions. Intravenous dobutamine infusion (24 mug. min-1. kg body wt-1) increased the mean heart rate by approximately 39% from 482 +/- 46 per min at baseline to 669 +/- 77 per min in adult mice. The myocardial phosphocreatine (PCr)-to-ATP (PCr/ATP) ratio remained unchanged at 2.1 +/- 0.5 during dobutamine stress, compared with baseline conditions. Therefore, we conclude that a significant increase in heart rate does not result in a decline in the in vivo murine cardiac PCr/ATP ratio. These observations in very small mammals, viz., mice, at extremely high heart rates are consistent with studies in large animals demonstrating that global levels of high-energy phosphate metabolites do not regulate in vivo myocardial metabolism during physiologically relevant increases in cardiac work.

Adenosine Triphosphate↗

High-energy phosphate responses to tachycardia and inotropic stimulation in left ventricular hypertrophy.

Spatially localized nuclear magnetic resonance (NMR) spectroscopy was used to examine the effect of tachycardia and inotropic stimulation on myocardial ATP, creatine phosphate (CrP), and inorganic phosphate (Pi) in animals with left ventricular hypertrophy (LVH). Studies were performed in eight normal dogs and seven dogs with moderate LVH produced by banding the ascending aorta. 31P-NMR spectra were obtained from five layers across the LV wall, while blood flow (BF) was measured with microspheres during control conditions, pacing at 200 and 240 beats/min, and during dobutamine infusion (Dob). Myocardial ATP and CrP levels were normal in the LVH hearts during control conditions. Pacing did not alter the transmural distribution of perfusion or the levels of CrP, ATP, and Pi in normal hearts. In contrast, in four of seven LVH hearts, pacing decreased the subendocardial/subepicardial (ENDO/EPI) BF ratio and caused depletion of CrP and appearance of Pi characteristic of ischemia in the subendocardium. Dob produced greater increases in the heart rate x LV systolic pressure product (RPP) and greater increases of Pi and decreases of CrP in LVH than in normal hearts; however, at comparable elevations of RPP the alterations of Pi and CrP were similar in both groups. Although Dob decreased the ENDO/EPI in LVH hearts, Dob-induced alterations in CrP and Pi were uniform across the LV wall. Increasing myocardial BF with adenosine or carbochromen did not reverse the alterations in Pi or CrP produced by Dob. We conclude that 1) ENDO perfusion abnormalities during tachycardia in LVH do produce ENDO subendocardial ischemia; 2) when the degree of augmentation of mechanical performance is considered, the metabolic changes induced by Dob were similar in normal and LVH hearts; 3) Dob-induced alterations in Pi and CrP were not related to inadequate perfusion, since increasing coronary BF did not reverse these changes; and 4) alterations of Pi and CrP during Dob infusion were not more prominent in the ENDO, indicating that the decreased ENDO/EPI flow did not cause ENDO ischemia but may reflect relatively lower O2 demands in this region during inotropic stimulation.

Adenosine Triphosphate↗

Hyperperfusion and cardioplegia effects on myocardial high-energy phosphate distribution and energy expenditure.

This study examines the hypothesis that high-energy phosphate (HEP) compound levels in unstimulated in vivo myocardium are defined by 1) the level of perfusion and 2) non-perfusion-dependent metabolic characteristics. This hypothesis was tested by determining 1) the effects of pharmacological hyperperfusion of functioning myocardium on transmural HEP compound distribution, contractile function, and myocardial oxygen consumption rate (MVO2) as well as 2) the effect of KCl cardioplegia on transmural myocardial HEP compound distribution. Creatine phosphate (CP) and ATP were measured across the anterior left ventricular wall using spatially localized 31P-nuclear magnetic resonance (NMR). At baseline, the CP-to-ATP (CP/ATP) ratio was significantly lower in the subendocardium than in the subepicardium. This transmural HEP gradient was abolished by hyperperfusion without significant effects on contractile function or MVO2. Similarly, KCl arrest significantly increased CP and CP/ATP in all myocardial layers, and the transmural gradient of CP/ATP was abolished again. These studies indicate that in present experimental model 1) myocardial performance is not constrained by inadequate perfusion in any myocardial layer although modest oxygen limitation affects the kinetics of oxidative phosphorylation in the inner myocardial layers and 2) in all myocardial layers, submaximal activation of intermediary metabolism and oxidative phosphorylation reactions results in lower steady-state CP and higher ADP levels relative to their respective values when energy expenditure is markedly reduced by KCl arrest.

Adenosine Triphosphate↗

Caffeine-induced Ca(2+) sparks in mouse ventricular myocytes.

Ca(2+) sparks are spatially localized intracellular Ca(2+) release events that were first described in 1993. Sparks have been ascribed to sarcoplasmic reticulum Ca(2+) release channel (ryanodine receptor, RyR) opening induced by Ca(2+) influx via L-type Ca(2+) channels or by spontaneous RyR openings and have been thought to reflect Ca(2+) release from a cluster of RyR. Here we describe a pharmacological approach to study sparks by exposing ventricular myocytes to caffeine with a rapid solution-switcher device. Sparks under these conditions have properties similar to naturally occurring sparks in terms of size and intracellular Ca(2+) concentration ([Ca(2+)](i)) amplitude. However, after the diffusion of caffeine, sparks first appear close to the cell surface membrane before coalescing to produce a whole cell transient. Our results support the idea that a whole cell [Ca(2+)](i) transient consists of the summation of sparks and that Ca(2+) sparks consist of the opening of a cluster of RyR and confirm that characteristics of the cluster rather than the L-type Ca(2+) channel-RyR relation determine spark properties.

Animals↗