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Motoneuron axon distribution in the cat stapedius muscle.

Stapedius-motoneuron cell bodies in the brainstem are spatially organized according to their acoustic response laterality, as demonstrated by intracellular labeling of physiologically identified motoneurons [Vacher et al., 1989. J. Comp. Neurol. 289, 401-415]. To determine whether a similar functional spatial segregation is present in the muscle, we traced physiologically identified, labeled axons into the stapedius muscle. Ten labeled axons were visible in the facial nerve and five could be traced to endplates within the muscle. These five axons had 39 observed branches (others may have been missed). This indicates an average innervation ratio (> or = 7.8) which is much higher than that obtained from previous estimates of the numbers of stapedius motoneurons and muscle fibers in the cat. One well-labeled stapedius motor axon innervated only a single muscle fiber. In contrast, two labeled axons had over 10 endings and innervated muscle fibers spread over wide areas in the muscle. Two of the axons branched and coursed through two primary stapedius fascicles, indicating that the muscle zones innervated by different primary fascicles are not functionally segregated. In another series of experiments, retrograde tracers were deposited in individual primary nerve fascicles. In every case, labeled stapedius-motoneuron cell bodies were found in each of the physiologically identified stapedius-motoneuron regions in the brainstem. These observations suggest there is little, if any, functional spatial segregation based on separate muscle compartments in the stapedius muscle, despite there being functional spatial segregation in the stapedius-motoneuron pool centrally.

Animals↗

[Studies on the spacial aerosol spreading during the use of turbines].

When treating hard tooth structures with a turbine instrument, a polydisperse aerosol is formed from the turbine exhaust air and the cooling-water spray and by the rotation of the abrasive tool. The oral cavity repels this aerosol that is contaminated with micro-organisms and particles of enamel and filling materials. The present paper deals with the spatial propagation of the aerosol cloud in front of the patient's mouth in relation to the site of impact of the turbine instrument. Recommendations are given for practice to reduce the possibilities of infection.

Aerosols↗

Tobacco RhoGTPase ACTIVATING PROTEIN1 spatially restricts signaling of RAC/Rop to the apex of pollen tubes.

Regulation by Rho-type small GTPases, such as RAC5, is important for the maintenance of polarity in tobacco (Nicotiana tabacum) pollen tubes. We previously showed that RhoGDI2 is necessary for RAC5 localization. Here, we describe the GTPase activating protein RhoGAP1 that controls the area of RAC5 activity. RhoGAP1 N-terminal and CRIB (for Cdc42/Rac-interactive binding) domains are both necessary for targeting yellow fluorescent protein-RhoGAP1 fusions to the plasma membrane close to, but not in, pollen tube apices. We propose that this localization restricts apical Rho-type GTPase activity from spreading toward the flanks, which ensures the maintenance of RAC signaling at the apex. The CRIB domain is not required but enhances in vitro RhoGAP1 activity toward the pollen tube-specific-RAC5. A mutation reducing GAP activity of RhoGAP1 leads to ballooning pollen tubes resembling those overexpressing RAC5. To ascertain the specific targeting mechanism of RhoGAP1, we isolated a 14-3-3 protein interacting with RhoGAP1. When overexpressed with RhoGAP1, it counteracts the growth-retarding effect of RhoGAP1 overexpression and attenuates RhoGAP1 membrane localization but, overexpressed alone, induces only small architectural changes. We propose that inactivation of RAC5 by the subapically localized RhoGAP1, together with dynamic relocalization of inactivated RAC5 from flanks to tip by RhoGDI2, leads to spatial restriction of RAC5 to pollen tube apices, thereby sustaining polar growth.

14-3-3 Proteins↗

Nonsynaptic epileptogenesis in the mammalian hippocampus in vitro. II. Role of extracellular potassium.

The role of extracellular K+ (K+o) in nonsynaptic epileptogenesis induced in the CA1 area of rat hippocampal slices by lowering [Ca2]o was studied with K+-selective microelectrodes (KSMs). Extracellular field potentials and [K+]o were recorded simultaneously with 1-2 KSMs in the CA1 stratum pyramidale. In slices perfused with an oxygenated standard physiological solution (containing 2 mM Ca2+), base-line [K+]o was stable for several hours. The washout of Ca2+o was accompanied by a gradual tonic rise of [K+]o. Spontaneous and stimulus-evoked maximal seizurelike events (SLEs) appeared when [K+]o was approximately 0.5 mM above the initial 5 mM base line. These changes were reversible in normal medium. When K+o was pressure ejected in the CA1 stratum pyramidale of spontaneously active slices, a local rise in [K+]o of approximately 0.5 mM was necessary to trigger a SLE. A similar apparent [K+]o "threshold" was associated with SLEs evoked by electrical stimulation. Increasing [K+] in the perfusing solution by small increments (1 mM) markedly enhanced SLEs frequency and velocity of spread and decreased the period of absolute refractoriness that succeeded each paroxysm. Similar changes occurred during periods of transient hypoxia. Small [K+] decreases in the perfusate had the converse effects. Spontaneous SLEs were associated with phasic increases in [K+]o. In simultaneous [K+]o recordings from two layers, these transients were largest (up to 3.5 mM above base line) and rose more steeply at the stratum pyramidale. Toward the outer dendritic layers they became smaller, slower in time course, and delayed in onset. We conclude that the main source for these [K+]o transients are the hippocampal pyramidal cell bodies, which discharge intensely during a SLE, and that excess K+o is spatially dispersed around the discharge zone of the paroxysm. [K+]o continued to rise, though at a slower rate, throughout the course of a SLE. Following SLE termination, [K+]o decayed slowly to base line. The invasion of a CA1 region by a propagating SLE was preceded quite often by a slow rise in [K+]o. A sudden transition to a steeply rising [K+]o marked the explosive recruitment of this region into the discharge zone of the spreading paroxysm. The total (tonic and phasic) increase in [K+]o during SLEs did not surpass a maximal level of approximately 9 mM, which was the ceiling level of [K+]o in low [Ca2+]o. However, when spreading depression occurred, [K+]o rose up to 30-40 mM for several minutes. Spreading depression rarely appeared spontaneously despite the recurrence of SLEs, but could be provoked by repetitive electrical stimulation.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Telodendrites of cone photoreceptors: structure and probable function.

Intracellular dye injection and compartmental modeling were used to analyze the structure and function of telodendrites of cones in the retina of the walleye. After identifying the spectral type of an impaled cone on the basis of its response to red and green light, horseradish peroxidase and/or Lucifer Yellow were injected for 1 to 25 minutes. In 38 of 58 recovered cells, dye spread into the telodendrites; so in many cases, the detailed pattern of the telodendritic arbor could be reconstructed from serial sections. Typically, five telodendritic processes, about 1 micron in diameter and 18 micron in length, radiated from the cone pedicle. A majority of the processes terminated at pedicles of neighboring cones. Some of the Lucifer Yellow injections provided evidence for electrical coupling between cones via telodendrites. Calculations from a compartmental model, based on the measured dimensions of cones and telodendrites, indicate that the signal arising in the inner segment spreads with little loss to the end of a telodendrite, whereas about half of the signal is lost in transmission from telodendrite to inner segment. Assuming that each contact point within the telodendritic network is an electrical synapse of 2,500 M omega, the model shows spatial interaction over a field of some 80 micron, which is comparable to that measured experimentally. Although our anatomical data indicate that orange- and green-sensitive cones may be interconnected via telodendrites, model calculations indicate that such connections do not appreciably distort the intrinsic spectral sensitivity of walleye cones. This outcome agrees with previous experimental results.

Animals↗

Time-independent point-spread function for NMR microscopy.

The resolution of NMR microscopy is analyzed in terms of the point-spread function, PSF(r), and the equivalent k space modulation transfer function, MTF(k). The analysis is developed for NMR spin warp and projection reconstruction imaging experiments; however, the framework provided is quite general. Incoherent spin motion is analyzed to predict what limits, if any, on spatial resolution are imposed by diffusion. Previous estimates of diffusion limits at 1-5 microns were developed for specific imaging techniques, typically using a mean displacement argument. Although qualitatively correct, the quantitative predictions represent practical rather than fundamental limits. It is shown that diffusion-dependent "blurring" can be made arbitrarily small and that the practical limits are less stringent than previously thought. A major point illustrated by the PSF-MTF formulation is that the irreversible loss of coherence by randomly diffusing spins occurs faster than the physical displacement, thereby reducing their effect considerably on the frequency or phase of the net detected signal. The irreversible loss of signal due to diffusive motion will contribute to and possibly dominate the signal-to-noise limit of resolution. The resolution as measured by the width of the PSF and MTF for diffusion is shown to be independent of the signal acquisition time, and their functional forms allow selection of microscopic imaging parameters. An example of a three-dimensional spin-warp image of a green algae cell is shown with resolution of approximately 16 microns x 13 microns x 10 microns.

Chlorophyta↗

Low-dispersion turns and junctions for microchannel systems.

Numerical methods are employed to optimize the geometry of two-dimensional microchannel turns such that the turn-induced spreading of a solute band is minimized. An inverted numerical method is first developed to compute the electric potential and local species motion in turns of arbitrary geometry. The turn geometry is then optimized by means of a nonlinear least-squares minimization algorithm using the spatial variance of the species distribution leaving the turn as the object function. This approach yields the turn geometry producing the minimum possible dispersion, subject only to prescribed constraints. The resulting low-dispersion turns provide an induced variance 2-3 orders of magnitude below that of a comparable conventional turns. Sample results are presented for 180 and 90 degrees turns, and the use of these turns to form wyes and tees is discussed. A sample 45 degrees wye is presented. The use of low-dispersion turns in folding separation columns is also discussed, and sample calculations are presented for folding a column 100 microm in width and up to 900 mm in length onto a region of only 10 by 10 mm. These low-dispersion geometries are applicable to electroosmosis, electrophoresis, and some pressure-driven flows.

Journal Article↗

Positron emission tomography within a magnetic field using photomultiplier tubes and lightguides.

The spatial resolution of positron emission tomography (PET) improves when positron annihilation takes place in a strong magnetic field. In a magnetic field, the Lorentz force restricts positron range perpendicular to the field. Since positron annihilation occurs closer to its point of origin, the positron annihilation point spread function decreases. This was verified experimentally by measuring the spread function of positron annihilation from a 500 mm 68Ge bead imbedded in tissue-equivalent wax. At 5 T the spread function full width at half maximum (FWHM) and the full width at tenth maximum (FWTM) decrease by a factor of 1.42 and 2.09, respectively. Two NaI(Tl) scintillation crystals that interface to a pair of photomultiplier tubes (PMTS) through long lightguides detect positron annihilation at zero field and 5.0 T. Photomultiplier tubes, inoperable in strong magnetic fields, are functional if lightguides bring the photons produced by scintillators within the field to a minimal magnetic field. These tests also demonstrate techniques necessary for combining magnetic resonance imaging (MRI) and PET into one scanner.

Gamma Rays↗

Intersymbol interference in underwater acoustic communications using time-reversal signal processing.

Coherent underwater communication is hampered by the time spread inherent to acoustic propagation in the ocean. Because time-reversal signal processing produces pulse compression, communications has been suggested as a natural application of the technique. Passive versions of time-reversal processing use a receive-only array to do combined temporal and spatial matched filtering. It can be shown, however, that the pulse compression it achieves is not perfect and that an equalizer that relies solely on time-reversal processing will have an error floor caused by uncompensated intersymbol interference (ISI). In the present paper, a physics-based model is developed for the uncompensated ISI in a passive time-reversal equalizer. The model makes use of a normal-mode expansion for the acoustic field. The matched-filtering integral is approximated and the intermediate result interpreted using the waveguide invariant. After combining across the array and sampling, formal statistical averages of the soft demodulation output are calculated. The results show how performance scales with bandwidth, with the number and position of array elements, and with the length of the finite impulse response matched filters. Good agreement is obtained between the predicted scaling and that observed in field experiments.

Journal Article↗

Particle-based multidimensional multispecies biofilm model.

In this paper we describe a spatially multidimensional (two-dimensional [2-D] and three-dimensional [3-D]) particle-based approach for modeling the dynamics of multispecies biofilms growing on multiple substrates. The model is based on diffusion-reaction mass balances for chemical species coupled with microbial growth and spreading of biomass represented by hard spherical particles. Effectively, this is a scaled-up version of a previously proposed individual-based biofilm model. Predictions of this new particle-based model were quantitatively compared with those obtained with an established one-dimensional (1-D) multispecies model for equivalent problems. A nitrifying biofilm containing aerobic ammonium and nitrite oxidizers, anaerobic ammonium oxidizers, and inert biomass was chosen as an example. The 2-D and 3-D models generally gave the same results. If only the average flux of nutrients needs to be known, 2-D and 1-D models are very similar. However, the behavior of intermediates, which are produced and consumed in different locations within the biofilm, is better described in 2-D and 3-D models because of the multidirectional concentration gradients. The predictions of 2-D or 3-D models are also different from those of 1-D models for slowly growing or minority species in the biofilm. This aspect is related to the mechanism of biomass spreading or advection implemented in the models and should receive more attention in future experimental studies.

Anaerobiosis↗

Risk analysis of the spread of classical swine fever virus through "neighbourhood infections" for different regions in Belgium.

Risk factors associated with the occurrence of "neighbourhood infections" [Epidemiology of classical swine fever. In: Truszczynski, M. (Ed.), Proceedings of the Workshop on Diagnostic Procedures and Measures to Control Classical Swine Fever in Domestic Pigs and the European Wild Boar. Pulaway, Poland, pp. 119-130] during classical swine fever (CSF) outbreaks were examined based on information collected during a CSF-epidemic, which occurred in the East Flanders Province of Belgium in 1994. The only risk factor that was associated with the occurrence of "neighbourhood infections" was a kernel estimation of the intensity of neighbouring herds (P=0.055) [Interactive spatial data analysis. Pearson Education Limited, Harlow, Essex], i.e. the higher the kernel estimation, the higher the risk for the occurrence of neighbourhood infections. In a second part of the study, the likelihood for the occurrence of neighbourhood infections within an area with a 1 km radius was predicted for every Belgian pig herd, assuming that the herd was infected with CSF-virus. For the prediction of these likelihoods, the model resulting from the risk assessment was used. Finally, the predicted likelihoods were transformed into a raster map after applying a smoothing technique. As a result, different areas in Belgium of higher or lower risk for CSF-virus spread through "neighbourhood infections" could be identified on the map. The areas in Belgium where CSF-outbreaks including "neighbourhood infections" occurred in the past decades were all predicted by the model to be of high risk.

Animals↗

[Magnetic resonance tomography compared to computed tomography in the intracardiac spread of hepatocarcinoma].

A cirrhotic patient with inferior cava and right atrium-ventricular invasion by hepatic tumor was studied with transthoracic echocardiography (TTE), computerized tomography (CT) before and after contrast agent bolus iv, ECG-gated magnetic resonance imaging (MRI). Obesity obstacled abdominal echotomographic study; esophageal varices were relative contraindication to transesophageal echocardiography (TEE). The resolutive diagnosis was possible thanks to MRI which documented the spatial continuity among hepatic tumor, intracaval neoplastic thrombus and intracardiac mass. TTE can be considered the first step in case of suspected cardiac masses, but it is sometimes limited by the thorax conformation and not always discriminant. TEE is semi-invasive with important contraindications. CT is limited by the only axial or para-axial scans with low quality reconstructions, worsened in these cases by respiratory and cardiac movements. MRI can be considered the second step in the imaging of cardiac masses, immediately and directly after the echocardiographic techniques.

Carcinoma, Hepatocellular↗

Molecular dissection of zyxin function reveals its involvement in cell motility.

Spatially controlled actin filament assembly is critical for numerous processes, including the vectorial cell migration required for wound healing, cell- mediated immunity, and embryogenesis. One protein implicated in the regulation of actin assembly is zyxin, a protein concentrated at sites where the fast growing ends of actin filaments are enriched. To evaluate the role of zyxin in vivo, we developed a specific peptide inhibitor of zyxin function that blocks its interaction with alpha-actinin and displaces it from its normal subcellular location. Mislocalization of zyxin perturbs cell migration and spreading, and affects the behavior of the cell edge, a structure maintained by assembly of actin at sites proximal to the plasma membrane. These results support a role for zyxin in cell motility, and demonstrate that the correct positioning of zyxin within the cell is critical for its physiological function. Interestingly, the mislocalization of zyxin in the peptide-injected cells is accompanied by disturbances in the distribution of Ena/VASP family members, proteins that have a well-established role in promoting actin assembly. In concert with previous work, our findings suggest that zyxin promotes the spatially restricted assembly of protein complexes necessary for cell motility.

Amino Acid Sequence↗

Focal adhesion kinase is required for the spatial organization of the leading edge in migrating cells.

The process of cell migration is initiated by protrusion at the leading edge of the cell, the formation of peripheral adhesions, the exertion of force on these adhesions, and finally the release of the adhesions at the rear of the cell. Focal adhesion kinase (FAK) is intimately involved in the regulation of this process, although the precise mechanism(s) whereby FAK regulates cell migration is unclear. We have used two approaches to reduce FAK expression in fibroblasts. Treatment of cells with FAK-specific siRNAs substantially reduced FAK expression and inhibited the spreading of fibroblasts in serum-free conditions, but did not affect the rate of spreading in the presence of serum. In contrast with the wild-type cells, the FAK siRNA-treated cells exhibited multiple extensions during cell spreading. The extensions appeared to be inappropriately formed lamellipodia as evidenced by the localization of cortactin to lamellipodial structures and the inhibition of such structures by expression of dominant-negative Rac. The wild-type phenotype was restored by reexpressing wild-type FAK in the knockdown cells, but not by expression of FAK containing a point mutation at the autophosphorylation site (FAK Y397F). In wound-healing assays, FAK knockdown cells failed to form broad lamellipodia, instead forming multiple leading edges. Similar results were obtained using primary mouse embryo fibroblasts from FAK-flox mice in which Cre-mediated excision was used to ablate the expression of FAK. These data are consistent with a role for FAK in regulating the formation of a leading edge during cell migration by coordinating integrin signaling to direct the correct spatial activation of membrane protrusion.

Animals↗

Reduced circular field-of-view imaging.

A rapid dynamic imaging technique based on polar k-space sampling is presented. A gain in temporal resolution is achieved by angular undersampling. A detailed analysis of the point spread function of angular undersampled polar imaging reveals a reduced diameter of the corresponding circular field of view. Under the assumption that dynamic changes are restricted to a local circular field of view, angular undersampled dynamic imaging allows the recording of rapid changes at high temporal and spatial resolution. The theoretical and experimental details of the technique are presented.

Artifacts↗

Evidence for social facilitation of preening in the common tern.

Social facilitation of reproductive behaviour has been studied extensively in gulls and terns, but social facilitation of preening has been reported only anecdotally, and has not been previously quantified. We studied a common tern, Sterna hirundo, colony during the summers of 1996 and 1997 to test for socially facilitated preening. Scan sampling provided evidence of spatial and temporal synchrony of preening behaviour. Preening occurred more often than expected in groups of three or more neighbours. Breeding pairs also preened simultaneously more often than expected. In loafing (resting) areas, the proportion of preeners present increased with tern density. Behavioural observations suggest that preening spread from neighbour to neighbour. The observed clumping in preening behaviour could not be explained by differences in date, time of day or weather. Social facilitation of preening and other maintenance behaviour may be an important aspect of group living that is often overlooked. Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗

The locus of interference in the Simon effect: an ERP study.

The Simon effect was studied using ERPs and the lateralized readiness potential (LRP). In Experiment 1, visual stimuli requiring left or right hand responses were presented at either side of fixation. In Experiment 2, the empty side was occupied by a distractor. Spatial S-R incompatibility delayed P300 latency in both experiments, as predicted by perceptual interference accounts of the Simon effect. However, the LRP results were not conclusive. The lateralized presentation of visual stimuli (Experiment 1) or targets (Experiment 2) produced a posterior ERP hemispheric asymmetry that spread to central electrodes and obscured the early development of the LRP. Experiment 3 avoided the problem by presenting the stimuli in the vertical meridian, and the LRP showed signs of incorrect response preparation in incompatible trials, in agreement with response-interference models. Both perceptual and response interference may be present in the Simon effect.

Adult↗

Transgenic reporter mouse for bioluminescence imaging of herpes simplex virus 1 infection in living mice.

Bioluminescence imaging allows spatial and temporal progression of viral infection to be detected and quantified in living mice, thereby providing a new approach for studies of viral-host pathogenesis. It has been necessary to construct and validate recombinant reporter viruses that express firefly luciferase to investigate viral replication and spread with this imaging technology. This strategy greatly limits the ability to analyze multiple strains of virus and/or existing viral mutants, and reporter viruses also may be attenuated relative to the respective parental viruses. To facilitate bioluminescence imaging of herpes simplex virus type 1 (HSV-1), we developed a transgenic reporter mouse that uses the promoter from HSV-1 thymidine kinase to control expression of firefly luciferase. Infection with HSV-1 activated expression of firefly luciferase in corneal and flank models of infection, and amounts of bioluminescence increased in proportion to increasing input titers of virus. Imaging could detect infection with three different strains of HSV-1 with the following relative rank order of bioluminescence produced at the site of infection: McKrae > 17 > KOS. Corneal infection with as few as 1 x 10(3) pfu strain McKrae was detectable above background levels. By comparison, infection with vaccinia virus did not affect bioluminescence in the reporter mouse. Collectively, these data establish a new transgenic reporter mouse for infection with HSV-1, thereby enabling in vivo bioluminescence imaging studies of HSV-1 pathogenesis without constructing new reporter viruses.

Animals↗