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Mitochondrial inhibition prior to oxygen-withdrawal facilitates the occurrence of hypoxia-induced spreading depression in rat hippocampal slices.

Oxygen withdrawal blocks mitochondrial respiration. In rat hippocampal slices, this triggers a massive depolarization of CA1 neurons and a negative shift of the extracellular DC potential, the characteristic sign of hypoxia-induced spreading depression (HSD). To unveil the contribution of mitochondria to the sensing of hypoxia and the ignition of HSD, we modified mitochondrial function. Mitochondrial uncoupling by carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP, 1 microM) prior to hypoxia hastened the onset and shortened the duration of HSD. Blocking mitochondrial ATP synthesis by oligomycin (10 microg/ml) was without effect. Inhibition of mitochondrial respiration by rotenone (20 microM), diphenyleneiodonium (25 microM), or antimycin A (20 microM) also hastened HSD onset and shortened HSD duration. 3-nitropropionic acid (1 mM) increased HSD duration. Cyanide (100 microM) hastened HSD onset and increased HSD duration. At higher concentrations, cyanide (1 mM), azide (2 mM), and FCCP (10 microM) triggered SD episodes on their own. Compared with control HSD, the spatial extent of the intrinsic optical signals of cyanide- and azide-induced SDs was more pronounced. Monitoring NADH (nicotinamide adenine dinucleotide) and FAD (flavin adenine dinucleotide) autofluorescence and mitochondrial membrane potential verified the mitochondrial targeting by the drugs used. Except 1 mM cyanide, no treatment reduced cellular ATP levels severely and no correlation was found between ATP, NADH, or FAD levels and the time to HSD onset. Therefore ATP depletion or a cytosolic reducing shift due to NADH/FADH2 accumulation cannot serve as a general explanation for the hastening of HSD onset on mitochondrial inhibition. Additional redox couples (glutathione) or events downstream of the mitochondrial depolarization need to be considered.

Adenosine Triphosphate↗

Distribution and spatial geometry of dopamine interplexiform cells in the rat retina: I. Developing retina.

The morphology and distribution of dopaminergic interplexiform cells were analyzed in 9-day-old rat retinas processed as wholemounts for tyrosine hydroxylase immunohistochemistry. The mean number of dopaminergic interplexiform cells was estimated as about half of the total number of dopaminergic neurons in the immature retina, with a higher density in the temporal retina. Four interplexiform cells were individually analyzed and reconstructed with a computer system. Their arborizations could be divided into three different regions based on both their morphological features and their position within the retinal layers: (1) an internal arborization, spreading at the margin between the inner nuclear layer and the inner plexiform layer, composed of long, thick, somatofugal dendrites branching at acute angles, (2) an external arborization in the middle of the inner nuclear layer, formed by short, thin, varicose, recurved, axon-like processes branching at right angles, (3) and one or more scleral process(es), originating either from the cell body or from the internal arborization, running toward the outermost cell row of the INL, some of which reached the outer plexiform layer. Finally, analysis of the arborization network by computer simulations based on the 4 digitalized cells was compared with a nearest-neighbour analysis of cell body distribution. It showed that cell bodies are almost randomly distributed--at least in the inferior retina--but that an adjustment of dendritic growth and orientation probably occurs to ensure a homogeneous coverage of the retina with a constant degree of overlap in the adult. This report represents the first three-dimensional computer reconstruction of chemically identified neurons in the retina.

Animals↗

[Spatial distribution and bloodmeal preferences of Glossina palpalis palpalis in the forest focus of Zoukougbeu (Ivory Coast). Epidemiological consequences].

In the sleeping sickness focus of Zoukougbeu (Côte d'Ivoire), in the cropping areas which are favourable for disease transmission, more than a quarter of the flies collected were found to have fed on domestic pigs. The sites where Glossina palpalis palpalis was caught fed on these animals were concordant with the sites where the patients were present. These results might indicate that in Zoukougbeu, but perhaps also in other sleeping sickness foci, the pig could play an active role in disease transmission, allowing the parasite to spread widely via the tsetse.

Animals↗

Echo train shifted multi-echo FLASH for functional MRI of the human brain at ultra-high spatial resolution.

This paper describes the development of a novel technique for functional MRI of the human brain at 0.135 microL resolution for a whole brain section. In comparison with conventional studies at 3 mm isotropic resolution or 27 microL voxel size, the method yields an improvement by a factor of 200. To achieve optimum image quality, the approach is based on a multi-echo fast low-angle shot (FLASH) sequence with unipolar traversals of k-space in the frequency-encoding dimension and echo train shifting to avoid amplitude discontinuities in the phase-encoding dimension. These strategies ensure a smooth point-spread function and eliminate image ghosting artifacts without the need for any phase correction or other post-processing. Signal-to-noise losses due to the considerably reduced voxel sizes are compensated for by single slice acquisitions, optimized bandwidths and an experimental four-channel shoulder coil matched to the posterior portion of the head. Multi-echo FLASH studies of the human brain (2.9 T, seven echoes, 200 Hz/pixel bandwidth, effective echo time 36 ms, acquisition time 6 s) at 300 microm resolution (no interpolation) and 1.5 mm slice thickness revealed robust activations in primary visual areas in response to binocular stimulation. The new method holds promise for refined studies of the columnar organization of specific brain systems and for functional assessments of the gray matter at laminar resolution.

Animals↗

Ecotoxicity assessment of natural attenuation effects at a historical dumping site in the western Baltic Sea.

During the late 1950s and early 1960s of the past century, industrial waste material highly enriched in various contaminants (heavy metals, PAHs) was dumped in the inner Mecklenburg Bay, western Baltic Sea. Large-scale shifts in the spatial distribution of heavy metals in surface sediments were mapped by geochemical monitoring in the mid-1980s and 12 years later in 1997. A further study in 2001 was designed to investigate the small-scale spatial distribution of contaminants inside, on top of, and around the historical dumping ground and to examine possible effects to benthic organisms (Arctica islandica, microbiological toxicity tests). The site is located within an area characterized by a discontinuous deposition of fine sediments, but net sedimentation rate of about 3mm/y led ultimately to a gradual coverage of the dumped material. During the first decades after the dumping, about half of the originally dumped material was already spread over the surrounding area as a result of sediment dynamics and re-suspension processes. Recent sediments of the historical dump site are still significantly enriched in heavy metals and PAHs. Microbiological toxicity test results were positive, though uptake of contaminants in mussels was found to deviate only slightly from that of a non-contaminated reference station due to a gradual dilution and decrease in bioavailability of the contaminants.

Animals↗

Stopping the motion and sleuthing the flash-lag effect: spatial uncertainty is the key to perceptual mislocalization.

A moving object is perceived to lie beyond a static object presented at the same time at the same retinal location (flash-lag effect or FLE). Some studies report that if the moving stimulus stops moving (flash-terminated condition or FTC) the instant the flash occurs, a FLE does not occur. Other studies, using different stimuli, report that the FLE does, in fact, occur in the FTC. The FTC is thus a crucial turning point in theories of flash-lag. Unraveling the mystery of the FLE in the FTC will help unravel the mechanisms underpinning flash-lag and perhaps even perceptual localization in general. Our experiments show that eccentricity of the moving stimulus was a contributing factor, as were eccentricity of the flashed stimulus and spatial separation between the two stimuli. Other factors, such as contrast and offset of moving stimulus, also modulate the magnitude of the FLE in the FTC. We surmise that uncertainty in determining the position in space of a moving stimulus is a key requirement for the lag-effect. A lag-effect in the FTC challenges influential models, such as differential latency, motion extrapolation, and postdiction. Based partly on the notion of an asymmetric spread of activity that arises because of the sheer nature of motion and from a combination of established physiological mechanisms, we propose a schematic account of the present findings that subsumes previous psychological models and scaffolds past experimental findings.

Contrast Sensitivity↗

Quantum dispersion and its exponential growth of a wave packet in chaotic systems.

The quantum correspondence of one particular signature of classical chaos--the exponential instability of motion--can be characterized by the initial exponential growth rate of the spreading of the propagating quantum wave packet. The growth rate is approximately twice the classical maximum Lyapunov exponent of the system. In the regular case, the dispersion of the wave packet is only due to the usual quantum effect that should vanish in the classical limit. In contrast, in the chaotic case, the evolution behavior of the wave packet is due to the dynamical effect associated with the nonlinearity and persists as long as the spatial extension of the initial wave packet is kept finite.

Journal Article↗

A method for modulation transfer function determination from edge profiles with correction for finite-element differentiation.

In this paper we describe a technique for determining the modulation transfer function (MTF) of an imaging system from an experimentally obtained edge profile. The technique includes an exact correction for the frequency passband of the finite-element differentiation required to obtain the line spread function from the edge spread function. This correction has been ignored by investigators in the past and is required whenever finite-element differentiation is used rather than analytic differentiation of a model fitted to the edge response data. The magnitude of the MTF correction is approximately 11% at f = fc/2 and approximately 57% at f = fc, where fc = fs/2 is the maximum frequency reproducible without aliasing with a sampling rate of fs. The correction is performed in the spatial frequency domain by multiplying the uncorrected MTF by 1/sinc (pi f/2fc). A computer simulation is presented to demonstrate the effect and the correction procedure. An experimental MTF of an x-ray image intensifier system obtained using this technique is found to be consistent with an MTF obtained using a bar pattern test phantom.

Computer Simulation↗

Changes in the distribution of LFA-1, catenins, and F-actin during transendothelial migration of monocytes in culture.

To determine changes in the spatial and temporal distribution of cell-cell adhesion molecules during transendothelial migration of monocytes, we examined an in vitro model system of diapedesis using high resolution laser scanning confocal microscopy. Human arterial endothelial cells were cultured to confluence on coverslips coated with Matrigel and activated with IL-1beta before the addition of monocytic THP-1 cells. Seventy per cent of monocytes transmigrated through the endothelium within one hour. Diapedesis, but not adhesion and spreading, was inhibited 8-fold in co-cultures that contained endothelial cell conditioned medium, suggesting the release of an endothelial derived inhibitor. Double immunofluorescence labeling with antibodies to LFA-1, alpha- and beta-catenin, VE-cadherin and with Texas Red phalloidin, identified a circular transmigration passage in endothelial cell-cell contact regions. This passage was formed by an LFA-1-containing pseudopodium that penetrated between endothelial cells. Apical to the transmigration passage, monocytes remained round in shape, while underneath the endothelium, they spread along the Matrigel. The margins of the transmigration passage contained high levels of LFA-1 and F-actin, suggesting a major role of these molecules during the transmigration process itself. Endothelial adherens junctions, as judged by the presence of VE-cadherin and alpha-catenin adjacent to the passage, remained intact during diapedesis. The presence of catenins at heterotypic contact regions between monocytes and endothelial cells during diapedesis suggested cadherin-mediated interactions between the two cell types. These results reveal dynamic changes in the distribution of adhesion molecules and the actin cytoskeleton during monocyte transendothelial migration in culture.

Actins↗

Current spread in the smooth muscle of the guinea-pig vas deferens.

1. The membrane polarization in response to intracellular stimulation and external stimulation, the junction potentials evoked by nerve and field stimulation and the spontaneous junction potentials were studied in the guinea-pig vas deferens.2. The responses to intracellular stimulation differed from those to external stimulation applied through a large electrode in the following ways: short time constant of the electrotonic potential; linearity of current-voltage relation; all-or-none spike only in a small proportion of the cells; high critical firing level; short latency; weak tendency for repetitive activity during depolarization; and sharp spatial decay of the response.3. The difference between intracellular and external stimulation could be explained by differences in current distribution in the tissue, if many muscle fibres were aggregated in functional bundles, with three-dimensional cell-to-cell connexions, so that the membrane near an intracellular stimulating electrode was shunted by a large area of surrounding membrane.4. The time course of the junction potentials depended on the manner by which they were produced. The junction potential evoked by hypogastric nerve stimulation was recorded in every cell with almost the same amplitude. The spontaneous junction potential decayed very sharply with distance and the time course of the falling phase was about 10 times faster than that of the evoked junction potential.The difference between the time course of the junction potentials was also explained by the difference in current distribution in the tissue.

Journal Article↗

Localization of subclinical ictal activity by functional magnetic resonance imaging: correlation with invasive monitoring.

Functional magnetic resonance imaging (fMRI) with susceptibility-based contrast was used to detect focal changes in cerebral blood flow and metabolism in a patient with focal epilepsy. The patient presented with frequent partial motor seizures involving his right lower face that spread to produce speech arrest and occasionally right arm jerking. Consciousness was never impaired during these events. A multislice echoplanar technique was used to acquire 16 contiguous axial slices every 4 seconds for 11 minutes. Although no overt seizures were observed or reported by the patient during the scanning, a time series analysis of the functional data revealed focal signal-intensity changes in the posterior left frontal lobe, which correlated well both in duration and spatial localization with ictal activity subsequently recorded by invasive electrophysiological monitoring. The spatial localization of fMRI was more accurate than electroencephalography recorded from a subdural grid in predicting the site of successful surgical therapy. These results illustrate the potential of functional MRI for localizing seizure foci with high spatial and temporal resolution. Such studies can be readily combined with high-resolution anatomical imaging, task-activation studies, and other magnetic resonance techniques.

Adult↗

Out-and-in spiral spectroscopic imaging in rat brain at 7 T.

With standard spectroscopic imaging, high spatial resolution is achieved at the price of a large number of phase-encoding steps, leading to long acquisition times. Fast spatial encoding methods reduce the minimum total acquisition time. In this article, a k-space scanning scheme using a continuous series of growing and shrinking, or "out-and-in," spiral trajectories is implemented and the feasibility of spiral spectroscopic imaging for animal models at high B(0) field is demonstrated. This method was applied to rat brain at 7 T. With a voxel size of about 8.7 microl (as calculated from the point-spread function), a 30 x 30 matrix, and a spectral bandwidth of 11 kHz, the minimum scan time was 9 min 20 sec for a signal-to-noise ratio of 7.1 measured on the N-acetylaspartate peak.

Animals↗

Diverse mechanisms of plant resistance to cauliflower mosaic virus revealed by leaf skeleton hybridization.

Plants not hosts for cauliflower mosaic virus (CaMV) may prevent systemic CaMV infection by interfering with dissemination of infection through the plant or by preventing viral replication and maturation. Leaf skeleton hybridization allows distinction between these two barriers. The technique assesses the spatial distribution of CaMV in an inoculated leaf by hybridization of a skeleton of the leaf with a CaMV DNA probe. Leaves or leaflets of soybean, cucumber, peanut, tomato, lettuce, spinach, pepper, onion, wheat, maize and barley, inoculated with CaMV DNA or CaMV virions were processed for leaf skeleton hybridization either immediately after inoculation or two weeks thereafter. Autoradiographic images of soybean and cucumber skeletons had many dark spots suggesting that CaMV DNA replication and local spread had occurred. Images of onion leaf skeletons prepared two weeks after inoculation with CaMV DNA had fewer spots. To test whether these spots resulted from CaMV replication, DNA was extracted from inoculated onion leaves and analyzed by electrophoresis, blotting and hybridization. Molecules recovered two weeks after inoculation resembled those inoculated, indicating absence of replication. For the other species, we found no evidence of local spread of CaMV infections. Thus, many plant species resist systemic CaMV infection by preventing replication or local spread of CaMV, while others solely prevent systemic movement of infection.

Blotting, Southern↗

A new method for comprehensive bird's-eye analysis of the surgically excised internal limiting membrane.

PURPOSE: To investigate the role of cells and extracellular matrix on the internal limiting membrane (ILM), we demonstrated a new method for a comprehensive bird's-eye analysis of surgically excised ILM. DESIGN: Laboratory investigation. METHODS: The ILM of an idiopathic epiretinal membrane was fixed and spread out onto a glass slide, using fine needles under a biomicroscope. The expanded ILM was analyzed by light microscopy, immunohistochemistry, and scanning electron microscopy. RESULTS: The excised ILM could be unfolded and spread out as a flat sheet onto the glass slide. Immunohistochemistry revealed that most migrating cells were glial cells. Scanning electronmicroscopy revealed that the vitreous surface of the ILM was smooth, in contrast to the rough retinal surface. The ILM showed abundant cellular migration as a continuous stratified cellular sheet. CONCLUSIONS: This new bird's-eye-view observation of excised ILM enables us to carry out comprehensive analysis of cellular distribution from a temporal and spatial perspective.

Basement Membrane↗

All-optical adaptive scanning acoustic microscope.

We have constructed a fast laser-based surface acoustic wave (SAW) microscope, which may be thought of as a non-perturbing scanning acoustic microscope. The instrument is capable of rapid high resolution vector contrast imaging at several discrete frequencies, without any damage to the sample. Tailoring the generating optical distribution using computer-generated holograms allows us to both focus the acoustic waves (increasing their amplitude) and to spread the optical power over the sample surface (preventing damage). Accurate quantitative amplitude and phase (velocity) measurements and unique acoustic contrast mechanisms are possible with our instrument based on this technology due to the non-perturbing nature and the instrument geometries.However, the complexity of the optical generation profile leads to a strong dependence on material properties such as the SAW velocity and material anisotropy. We address these issues in this paper, and demonstrate how a spatial light modulator may be used to adapt the generating optical distribution to compensate for the material properties. This facilitates simpler alignment and velocity matching, and, combined with an acoustic wavefront sensor, will allow real-time adjustment of the generating source to enable imaging on anisotropic materials.

Equipment Design↗

Three-dimensional analysis of mineralizing turkey leg tendon: matrix vesicle-collagen relationships.

The spatial and temporal relationships of mineral deposition between matrix vesicles and type I collagen fibrils have been studied in the turkey leg tendon by electron microscopy of cross sections and serial longitudinal thin sections and by electron tomography of longitudinal thick sections. Serial sectioning and electron tomography allow three-dimensional analysis of spatial relationships, overcoming the problems of missing depth information and over-projection of adjacent structures which exist for two-dimensional projections of isolated sections. These techniques reveal that while mineral deposits within matrix vesicles are found remote from calcifying collagen fibrils, the reverse relationship does not occur; all collagen-associated mineral can ultimately be linked to mineral-laden vesicles. These results suggest a temporal sequence of calcification beginning in matrix vesicles and spreading to adjacent collagen fibrils.

Animals↗

The effects of interpulse interval on stochastic properties of electrical stimulation: models and measurements.

It is known that some cochlear implant users have improved speech perception using higher rates of interleaved pulsatile stimulation. There are, however, significant limitations on their performance presumably due in part to temporal and spatial interactions. To address these limitations, we have examined refractory characteristics of the auditory nerve using experimental animal models and computational simulations. A stochastic model of the node of Ranvier modified for mammalian sodium channel kinetics has been developed to calculate the masked input-output (I/O) functions for different interpulse intervals (IPI) [26]. The model is based upon 1000 voltage-gated sodium channels and incorporates parameters such as nodal resistance and capacitance. The relative spread (RS) [35] calculated from the I/O functions was typically 0.03 for 17 different IPIs between 450 micros and 6 ms for cathodal stimuli. For IPI = 830 and 870 micros, the RS was ten times greater than those for other IPIs. Although it is not fully understood how the electrically evoked compound action potential (EAP) data are related to single fiber data, the RS of single fibers is a partial contributor [19]. We have measured the EAP using a monopolar intracochlear stimulating electrode and a recording electrode placed directly on the nerve and have observed changes in slope of EAP growth functions consistent with the theoretical RS values. These results have significant implications for speech coding in a cochlear implant since they suggest an increased membrane noise for pulse trains of specific rates.

Action Potentials↗

Age-dependent poliomyelitis of mice: expression of endogenous retrovirus correlates with cytocidal replication of lactate dehydrogenase-elevating virus in motor neurons.

The widespread presence of endogenous retroviruses in the genomes of animals and humans has suggested that these viruses may be involved in both normal and abnormal developmental processes. Previous studies have indicated the involvement of endogenous ecotropic murine leukemia virus (MuLV) in the development of age-dependent poliomyelitis caused by infection of old C58 or AKR mice by lactate dehydrogenase-elevating virus (LDV). The only genetic components which segregate with susceptibility to LDV-induced paralytic disease are multiple proviral copies of ecotropic MuLV and the permissive allele, at the Fv-1 locus, for N-tropic, ecotropic virus replication (Fv-1n/n). Using in situ hybridization and Northern (RNA) blot hybridization, we have correlated the expression of the endogenous MuLV, both temporally and spatially, with LDV infection of anterior horn motor neurons and the development of paralysis. Our data indicate that treatment of 6- to 7-month-old C58/M mice with cyclophosphamide, which renders these mice susceptible to LDV-induced paralytic disease, results in transient increases in ecotropic MuLV RNA levels in motor neurons throughout the spinal cord. Peripheral inoculation of C58/M mice with LDV, at the time of elevated MuLV RNA levels, results in a rapid spread of LDV to some spinal cord motor neurons. LDV infections then spread slowly but progressively throughout the spinal cord, involving an increasing number of motor neurons. LDV replication is cytocidal and results in neuron destruction and paralysis of the infected animals 2 to 3 weeks postinfection. The slow replication of LDV in the spinal cord contrasts sharply with the rapid replication of LDV in macrophages, the normal host cells for LDV, during the acute phase of infection. The data indicate that the interaction between the endogenous MuLV with the generally nonpathogenic murine togavirus LDV occurs at the level of the motor neuron. We discuss potential mechanisms for the novel dual-virus etiology of age-dependent poliomyelitis of mice.

Age Factors↗