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Regulation of exocytosis via release of Ca(2+) from intracellular stores.

The release of Ca(2+) from intracellular stores is an important trigger for secretion in many cell types. Depending on the spatial relationship between the intracellular Ca(2+) stores and the site of exocytosis, the Ca(2+) signal can be very local or spread throughout the entire cell. Here, we review how the release of Ca(2+) from inositol trisphospate (IP(3))-sensitive stores contributes differently to the stimulus-secretion coupling in three types of secretory cells (acinar cells of the pancreas, gonadotrophs, and corticotrophs of the anterior pituitary gland). We propose that in both pancreatic acinar cells and pituitary gonadotrophs the IP(3)-sensitive stores may be in close proximity to the sites of exocytosis such that the concentration of Ca(2+) at these sites are transiently much higher than the average cytosolic Ca(2+) concentration. In contrast, the local Ca(2+) gradient is less prominent in pituitary corticotrophs. Finally, some recent technical developments that may contribute significantly to future investigations of local Ca(2+) signals are discussed.

Adrenocorticotropic Hormone↗

Electrical coupling between glial cells in the rat retina.

The strength of electrical coupling between retinal glial cells was quantified with simultaneous whole-cell current-clamp recordings from astrocyte-astrocyte, astrocyte-Müller cell, and Müller cell-Müller cell pairs in the acutely isolated rat retina. Experimental results were fit and space constants determined using a resistive model of the glial cell network that assumed a homogeneous two-dimensional glial syncytium. The effective space constant (the distance from the point of stimulation to where the voltage falls to 1/e) equaled 12.9, 6.2, and 3.7 microm, respectively for astrocyte-astrocyte, astrocyte-Müller cell, and Müller cell-Müller cell coupling. The addition of 1 mM Ba(2+) had little effect on network space constants, while 0.5 mM octanol shortened the space constants to 4.7, 4.4, and 2.6 microm for the three types of coupling. For a given distance separating cell pairs, the strength of coupling showed considerable variability. This variability in coupling strength was reproduced accurately by a second resistive model of the glial cell network (incorporating discrete astrocytes spaced at varying distances from each other), demonstrating that the variability was an intrinsic property of the glial cell network. Coupling between glial cells in the retina may permit the intercellular spread of ions and small molecules, including messengers mediating Ca(2+) wave propagation, but it is too weak to carry significant K(+) spatial buffer currents.

Animals↗

Imaging of finger joints in a whole-body MR system using a simple and low-cost solenoidal coil.

This work considers the design and operation of a solenoid radiofrequency (RF) coil for magnetic resonance (MR) mini-imaging of finger joints using a whole-body system. The considerations were focused on the requirements of high sensitivity, easy patient access, and cost-effectiveness. The proposed design is a short, two-turn solenoidal coil that accomplishes high spatial uniformity over a 2-cm region of interest (ROI). The coil resembles an oversized ring, and easily fits fingers that are spread. An experimental demonstration of the proposed approach is given by imaging the proximal interphalangeal joint in healthy volunteers using a 2.5-cm field of view (FOV). Spin-echo and gradient-recalled-echo T1-weighted sequences, and a 3D spoiled-gradient-echo sequence were used.

Cost-Benefit Analysis↗

An iterative method to calculate forces exerted by single cells and multicellular assemblies from the detection of deformations of flexible substrates.

We present a new method for quantification of traction forces exerted by migrating single cells and multicellular assemblies from deformations of flexible substrate. It is based on an iterative biconjugate gradient inversion method. We show how the iteration and the solution are influenced by experimental parameters such as the noise on deformations sigma ( XY ), and the mean depth of recorded deformations Z (M). In order to find the validity range of our computational method, we simulated two different patterns of force. The first artificial force pattern mimics the forces exerted by a migrating Dictyostelium slug at a spatial resolution of Delta=20 mum (Rieu et al. in Biophys J 89:3563-3576, 2005) and corresponds to a large and spread force field. The second simulated force pattern mimics forces exerted by a polarized fibroblast at discrete focal adhesion sites separated by Delta=4 microm. Our iterative method allows, without using explicit regularization, the detailed reconstruction of the two investigated patterns when noise is not too high (sigma ( XY )/u (max)< or =6%, where u (max )is the maximal deformation), and when the plane of recorded deformations is close to the surface (Delta/Z (M)> or =4). The method and the required range of parameters are particularly suitable to study forces over large fields such as those observed in multicellular assemblies.

Algorithms↗

Secretagogue-induced calcium wave shows higher and prolonged transients of nuclear calcium concentration in mast cells.

To clarify the mechanism of secretagogue (compound 48/80)-induced calcium signaling in rat peritoneal mast cells, we analyzed serial confocal calcium images with high spatial and temporal resolution using different Ca(2+)-probes. The Ca(2+)-wave began at the periphery of the cytoplasm, and then spread to the center of the nucleus. Nuclear [Ca2+]i was clearly higher than cytoplasmic [Ca2+]i. The heterogeneity of [Ca2+]i continued until about 2 min after degranulation. These results suggest the existence of an intranuclear Ca(2+)-store which possesses a Ca(2+)-releasing mechanism similar to that in the cytoplasm.

Animals↗

Secondary tumor formation after stereotactic biopsy for intracerebral metastatic disease.

BACKGROUND: There are only a few published cases of iatrogenic tumor seeding and distant neoplastic growth along the path of the cannula after stereotactic biopsy. CASE DESCRIPTION: We report the case of a female patient with a solitary lung cancer metastasis in the left parietal brain lobe. The tumor was stereotactically biopsied and treated by radiosurgery. One month after the initial biopsy, a smaller de novo tumor mass located along the track of the stereotactic cannula was detected by contrast-enhanced MRI. The spatial location of this neoplastic nodule and the short time before development of a macroscopic lesion seemed to confirm iatrogenic tumor cell spread due to the stereotactic procedure. CONCLUSION: Our findings and the reviewed literature suggest that this complication is rare. Nevertheless, neurosurgeons should be aware of the potential risk and, if necessary, should be able to modify the technical procedure and the adjuvant treatment.

Biopsy, Needle↗

Phase contrast MRI with improved temporal resolution by view sharing: k-space related velocity mapping properties.

Phase contrast techniques in combination with k-space segmented CINE imaging are widely used for the quantitative assessment of blood flow or tissue motion. The temporal resolution of the corresponding pulse sequences plays an important role concerning the potential of the method to fully detect time resolved flow or motion patterns. A further improvement of temporal or spatial resolution in phase contrast CINE MRI can be achieved by the application of view sharing. Based on simulations with point-spread-functions resulting from different cyclic flow or motion patterns an analysis of view sharing techniques in combination with phase contrast MRI is presented. Velocity mapping properties and the role of different k-space regions concerning the resulting values in the phase images and thus encoded velocities were investigated. It could be shown that the velocity induced phase shifts in phase contrast techniques are mainly encoded in the central sections of k-space which makes view sharing also suitable for velocity mapping. As a result the use of appropriate sampling and data acquisition schemes permits the assessment of flow or motion patterns with significantly improved temporal resolution without loss of functional information. In addition phantom measurements with an oscillation phantom were performed in order to validate the simulation results and to demonstrate the potential of view sharing techniques to accelerate phase contrast imaging and improve the detection of the underlying flow or motion dynamics.

Blood Flow Velocity↗

Principles of high-spatial-resolution surface EMG (HSR-EMG): single motor unit detection and application in the diagnosis of neuromuscular disorders.

The most detailed information about the structural and functional characteristics of the muscle can be gained from the single motor unit (MU) action potential. In addition, information about the activity of a single MU is essential for the diagnosis of neuromuscular disorders. Due to the low spatial resolution of conventional bipolar surface electromyography (EMG), the resulting signal is a superposition of a large number of simultaneous active MUs. The difficulty is in separating the activity of a single MU from simultaneous active adjacent MUs. In contrast to other non-invasive EMG procedures, the high-spatial-resolution-EMG (HSR-EMG), which is based on the use of a multi-electrode array in combination with a spatial filter procedure, allows the detection of single MU activity in a non-invasive way. It opens access to the excitation spread and enables the determination of the conduction velocity in single MUs, and the localization of the endplate region. In addition, HSR-EMG detects changes in the electrical activities of the MUs which are typical in neuromuscular disorders. Using HSR-EMG it was possible to identify 97% of all investigated volunteers and patients with muscular or neuronal disorders. Therefore, HSR-EMG is suitable as a tool for the non-invasive diagnosis of neuromuscular disorders.

Journal Article↗

One-dimensional rabbit sinoatrial node models: benefits and limitations.

INTRODUCTION: Cardiac multicellular modeling has traditionally focused on ventricular electromechanics. More recently, models of the atria have started to emerge, and there is much interest in addressing sinoatrial node structure and function. METHODS AND RESULTS: We implemented a variety of one-dimensional sinoatrial models consisting of descriptions of central, transitional, and peripheral sinoatrial node cells, as well as rabbit or human atrial cells. These one-dimensional models were implemented using CMISS on an SGI Origin 2000 supercomputer. Intercellular coupling parameters recorded in experimental studies on sinoatrial node and atrial cell-pairs under-represent the electrotonic interactions that any cardiomyocyte would have in a multidimensional setting. Unsurprisingly, cell-to-cell coupling had to be scaled-up (by a factor of 5) in order to obtain a stable leading pacemaker site in the sinoatrial node center. Further critical parameters include the gradual increase in intercellular coupling from sinoatrial node center to periphery, and the presence of electrotonic interaction with atrial cells. Interestingly, the electrotonic effect of the atrium on sinoatrial node periphery is best described as opposing depolarization, rather than necessarily hyperpolarizing, as often assumed. CONCLUSION: Multicellular one-dimensional models of sinoatrial node and atrium can provide useful insight into the origin and spread of normal cardiac excitation. They require larger than "physiologic" intercellular conductivities in order to make up for a lack of "anatomical" spatial scaling. Multicellular models for more in-depth quantitative studies will require more realistic anatomico-physiologic properties.

Action Potentials↗

Space charge effects in a kinestatic charge detector.

The effects of space charge in a kinestatic charge detector (KCD) were examined using computer solutions to Poisson's equation. The KCD is a strip-beam parallel-plate drift chamber used for digital radiography. It was assumed that there is negligible electron attachment, i.e. there are no negative ions formed. The ionization rate per mA as a function of x-ray interaction depth was calculated for a detector filled with xenon at 25.3 x 10(5) Pa. Solution of Thomson's equations gave the positive ion density at the cathode, also as a function of depth. Water filtration values ranging from 0 to 30 cm were used in order to estimate the range of ion density values expected in a clinical KCD. The case of steady-state x-ray illumination was simulated for ionization rates less than the zero field limit (above which space charge changes the polarity of the electric field). Line spread responses were found for varying ionization rates to show the effect of space charge due to electric field distortion on the spatial resolution performance in the drift direction. The effect of imaging ideal edges with a KCD was calculated and the expected output signal was plotted for densities up to the zero field limit. Space charge dependence on the selection of KCD design and operating parameters is discussed. Because of the dependence of the KCD drift-direction spatial resolution on the uniformity of the electric field, space charge effects impose an upper limit on the detector entrance exposure and define the dynamic range of the device.

Humans↗

Fundamental clusters in spatial 2x2 games.

The notion of fundamental clusters is introduced, serving as a rule of thumb to characterize the statistical properties of the complex behaviour of cellular automata such as spatial 2 x 2 games. They represent the smallest cluster size determining the fate of the entire system. Checking simple growth criteria allows us to decide whether the cluster-individuals, e.g. some mutant family, are capable of surviving and invading a resident population. In biology, spatial 2 x 2 games have a broad spectrum of applications ranging from the evolution of cooperation and intraspecies competition to disease spread. This methodological study allows simple classifications and long-term predictions in various biological and social models to be made. For minimal neighbourhood types, we show that the intuitive candidate, a 3 x 3 cluster, turns out to be fundamental with certain weak limitations for the Moore neighbourhood but not for the Von Neumann neighbourhood. However, in the latter case, 2 x 2 clusters generally serve as reliable indicators to whether a strategy survives. Stochasticity is added to investigate the effects of varying fractions of one strategy present at initialization time and to discuss the rich dynamic properties in greater detail. Finally, we derive Liapunov exponents for the system and show that chaos reigns in a small region where the two strategies coexist in dynamical equilibrium.

Animals↗

Calcium clamp in isolated neurones of the snail Helix pomatia.

1. Intracellular free calcium concentration ([Ca2+]i) in isolated non-identified Helix pomatia neurones has been clamped at different physiologically significant levels by a feedback system between the fluorescent signal of fura-2 probe loaded into the cell and ionophoretic injection of Ca2+ ions through a CaCl2-loaded microelectrode. The membrane potential of the neurone has also been clamped using a conventional two-microelectrode method. 2. Special measurements have shown that the transport indices of injecting microelectrodes filled with 50 mM CaCl2 are quite variable (0.11 +/- 0.06, mean +/- S.D.). However, for each electrode the transport indices remained stable during several injection trials into a solution drop having the size of a neurone. The spread of calcium ions from the tip of the microelectrode across the cytosol of the neurone terminated within 2-4 s. The spatial difference in [Ca2+]i at this time did not exceed 10%. 3. Clamping of [Ca2+]i at a new increased level was accompanied by a transient of the Ca(2+)-injecting current. To increase [Ca2+]i by 0.1 microM, the amount of calcium ions injected during this stage had to be 36 +/- 20 microM Ca2+ per cell volume. Obviously, this transient represents the filling of a fast cytosolic buffer which has to be saturated to reach a new increased level of [Ca2+]i. It was followed by a steady component of Ca(2+)-injecting current, which was quite low (corresponding to injection of 0.39 +/- 0.20 microM s-1 for a 0.1 microM change of [Ca2+]i). This may represent the functioning of Ca(2+)-eliminating systems and corresponds to a similar amount of Ca2+ extruded from the cytoplasm. 4. Changes in the injection current also developed when Ca2+ influx through the membrane was triggered by the activation of voltage-gated calcium channels. The amount of Ca2+ entering the cell during the first seconds of depolarization to--15 mV was equal to 0.59 +/- 0.31 microM s-1 per cell volume. 5. No activation of Ca(2+)-dependent potassium current was observed during the changes in [Ca2+]i to levels exceeding the basal one by several times. Obviously, to activate this current, a much stronger increase in [Ca2+]i is needed in the immediate vicinity of the corresponding channels.

Animals↗

Use of a magnetic field to increase the spatial resolution of positron emission tomography.

Detector geometry, spatial sampling, and more fundamentally, positron range and noncollinearity of annihilation photon emission define Positron Emission Tomography (PET) spatial resolution. In this paper, a strong magnetic field is used to constrain positron travel transverse to the field. Measurement of the spread function from a 500 microns diameter 68Ga impregnated resin bead shows a squeezing of the full width at half maximum (FWHM) by a factor of 1.0, 1.22, 1.42, and 2.05, at 0, 4.0, 5.0, and 9.4 Tesla, respectively. The full width at tenth maximum (FWTM) decreases by a factor of 1.0, 1.73, 2.09, and 3.20, at 0, 4.0, 5.0, and 9.0 Tesla, respectively. Acquiring a PET image in a magnetic field should significantly reduce resolution loss due to positron range.

Biophysical Phenomena↗

Comparison of matched BOLD and FAIR 4.0T-fMRI with [15O]water PET brain volumes.

Valid comparisons of functional activation volumes from fMRI and PET require accurate registration, matched spatial resolution, and if possible matched noise. We coregistered 4.0T-fMRI and PET volumes, using a series of linear and nonlinear transformations applied to the PET volumes. Because of the limited number of fMRI slices that were available, PET volumes were transformed to the fMRI space. Since 4.0T-fMRI and 4.0T-MRI volumes have significant spatial distortion due to magnet inhomogeneities, high resolution 1.5T-MRI volumes were nonlinearly transformed to 4.0T-MRI volumes as part of the transformation chain. The smoothing effects of these registration transformations were measured, in order to match the spatial resolution of the coregistered fMRI and PET volumes. Spatial resolution of the transformed PET volumes in the fMRI space was degraded by up to 60% due to the transformation process. Due to both the image acquisition characteristics and the coregistration process, the transformed PET volumes had a spatial resolution that was lower than that of tMRI. Therefore, significant smoothing of fMRI volumes was necessary to match their spatial resolution with that of the transformed PET volumes. Matching the spatial resolution of the fMRI volumes to those of the transformed PET volumes was achieved by matching the shape of their point spread functions. In order to do this, Gaussian kernels were employed to smooth the fMRI volumes. We were unable to simultaneously match the resolution and noise of fMRI and PET signals in the motor cortex. Activation maps derived from transformed PET and smoothed fMRI volumes were compared. Contralateral motor cortex was active in all modalities but there were large variations in the size of the activated region and its signal to noise ratio across BOLD, FAIR, and PET images within each subject. Nevertheless, the relative CBF changes measured by FAIR were consistent with those determined by PET.

Biophysical Phenomena↗

A method for deconvolution of integrated electronic portal images to obtain incident fluence for dose reconstruction.

A method to convert integrated electronic portal imaging device (EPID) images to fluence for the purpose of reconstructing the dose to a phantom is investigated here for simple open fields. Ultimately, the goal is to develop a method to reconstruct the dose to patients. The EPID images are transformed into incident intensity fluence by spatial filtering with a deconvolution kernel. The kernel uses a general mathematical form derived from a Monte Carlo calculation of the point spread function of an EPID. The deconvolution kernel is fitted using a downhill search algorithm that minimizes the difference between the reconstructed dose and the dose measured in water. The beam profile "horns" that are removed by the EPID calibration procedure are restored to the resulting images by direct multiplication using the measured in-air off-axis ratio. Applying the fitted kernel to an EPID image provides the incident fluence for that beam. This beam fluence is then entered into an independent dose calculation algorithm for phantom or patient dose reconstruction. The phantom dose was computed to an accuracy of 2.0% of the dmax dose at one standard deviation. The method is general and can possibly be applied to any EPID equipped with an integration mode. We demonstrate the application of the fitted kernel in two clinical IMRT cases.

Algorithms↗

Physiological imaging of the lung: single-photon-emission computed tomography (SPECT).

Emission tomography provides three-dimensional, quantitative images of the distribution of radiotracers used to mark physiological, metabolic, or pathological processes. Quantitative single photon emission computed tomography (SPECT) requires correction for the image-degrading effects due to photon attenuation and scatter. Phantom experiments have shown that radioactive concentrations can be assessed within some percentage of the true value when relevant corrections are applied. SPECT is widely spread, and radiotracers are available that are easy to use and comparably inexpensive. Compared with other methods, SPECT suffers from a lower spatial resolution, and the time required for image acquisition is longer than for some alternative methods. In contrast to some other methods, SPECT allows simultaneous imaging of more than one process, e.g., both regional blood flow and ventilation, for the whole lung. SPECT has been used to explore the influence of posture and clinical interventions on the spatial distribution of lung blood flow and ventilation. Lung blood flow is typically imaged using macroaggregates of albumin. Both radioactive gases and particulate aerosols labeled with radioactivity have been used for imaging of regional ventilation. However, all radiotracers are not equally suited for quantitative measurements; all have specific advantages and limitations. With SPECT, both blood flow and ventilation can be marked with radiotracers that remain fixed in the lung tissue, which allows tracer administration during conditions different from those at image registration. All SPECT methods have specific features that result from the used radiotracer, the manner in which it is administered, and how images are registered and analyzed.

Animals↗

Binary image restoration by positive semidefinite programming.

We report an optimization approach to restore degraded binary images by using positive semidefinite programming when the point spread function (PSF) is known. The approach takes advantage of the combinatorial nature of the problem, considering not only local similarity and spatial context but also the relationship between individual pixel values and the PSF. Numerical experiments confirm the superiority of the approach.

Journal Article↗

Mitochondrial calcium ion and membrane potential transients follow the pattern of epileptiform discharges in hippocampal slice cultures.

Emerging evidence suggests that mitochondrial dysfunction contributes to the pathophysiology of epilepsy. Recurrent mitochondrial Ca2+ ion load during seizures might act on mitochondrial membrane potential (DeltaPsim) and proton motive force. By using electrophysiology and confocal laser-scanning microscopy, we investigated the effects of epileptiform activity, as induced by low-Mg2+ ion perfusion in hippocampal slice cultures, on changes in DeltaPsim and in mitochondrial Ca2+ ion concentration ([Ca2+]m). The mitochondrial compartment was identified by monitoring DeltaPsim in the soma and dendrites of patched CA3 pyramidal cells using the mitochondria-specific voltage-sensitive dye rhodamine-123 (Rh-123). Interictal activity was accompanied by localized mitochondrial depolarization that was restricted to a few mitochondria in small dendrites. In contrast, robust Rh-123 release into the cytosol was observed during seizure-like events (SLEs), indicating simultaneous depolarization of mitochondria. This was critically dependent on Ca2+ ion uptake and extrusion, because inhibition of the mitochondrial Ca2+ ion uniporter by Ru360 and the mitochondrial Na+/Ca2+ ion exchanger by 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3H)-one but not the inhibitor of mitochondrial permeability transition pore, cyclosporin A, decreased the SLE-associated mitochondrial depolarization. The Ca2+ ion dependence of simultaneous mitochondrial depolarization suggested enhanced Ca2+ ion cycling across mitochondrial membranes during epileptiform activity. Indeed, [Ca2+]m fluctuated during interictal activity in single dendrites, and these fluctuations spread over the entire mitochondrial compartment during SLEs, as revealed using mitochondria-specific dyes (rhod-2 and rhod-ff) and spatial frequency-based image analysis. These findings strengthen the hypothesis that epileptic activity results in Ca2+ ion-dependent changes in mitochondrial function that might contribute to the neuronal injury during epilepsy.

Animals↗