The role of blood vessels in high-resolution volume conductor head modeling of EEG.

Reconstruction of the electrical sources of human EEG activity at high spatio-temporal accuracy is an important aim in neuroscience and neurological diagnostics. Over the last decades, numerous studies have demonstrated that realistic modeling of head anatomy improves the accuracy of source reconstruction of EEG signals. For example, including a cerebro-spinal fluid compartment and the anisotropy of white matter electrical conductivity were both shown to significantly reduce modeling errors. Here, we for the first time quantify the role of detailed reconstructions of the cerebral blood vessels in volume conductor head modeling for EEG. To study the role of the highly arborized cerebral blood vessels, we created a submillimeter head model based on ultra-high-field-strength (7T) structural MRI datasets. Blood vessels (arteries and emissary/intraosseous veins) were segmented using Frangi multi-scale vesselness filtering. The final head model consisted of a geometry-adapted cubic mesh with over 17×10(6) nodes. We solved the forward model using a finite-element-method (FEM) transfer matrix approach, which allowed reducing computation times substantially and quantified the importance of the blood vessel compartment by computing forward and inverse errors resulting from ignoring the blood vessels. Our results show that ignoring emissary veins piercing the skull leads to focal localization errors of approx. 5 to 15mm. Large errors (>2cm) were observed due to the carotid arteries and the dense arterial vasculature in areas such as in the insula or in the medial temporal lobe. Thus, in such predisposed areas, errors caused by neglecting blood vessels can reach similar magnitudes as those previously reported for neglecting white matter anisotropy, the CSF or the dura - structures which are generally considered important components of realistic EEG head models. Our findings thus imply that including a realistic blood vessel compartment in EEG head models will be helpful to improve the accuracy of EEG source analyses particularly when high accuracies in brain areas with dense vasculature are required.

Variability in growth/no growth boundaries of 188 different Escherichia coli strains reveals that approximately 75% have a higher growth probability under low pH conditions than E. coli O157:H7 strain ATCC 43888.

This study investigated the variation in growth/no growth boundaries of 188 Escherichia coli strains. Experiments were conducted in Luria-Bertani media under 36 combinations of lactic acid (LA) (0 and 25 mM), pH (3.8, 3.9, 4.0, 4.1, 4.2 and 4.3 for 0 mM LA and 4.3, 4.4, 4.5, 4.6, 4.7 and 4.8 for 25 mM LA) and temperature (20, 25 and 30 °C). After 3 days of incubation, growth was monitored through optical density measurements. For each strain, a so-called purposeful selection approach was used to fit a logistic regression model that adequately predicted the likelihood for growth. Further, to assess the growth/no growth variability for all the strains at once, a generalized linear mixed model was fitted to the data. Strain was fitted as a fixed factor and replicate as a random blocking factor. E. coli O157:H7 strain ATCC 43888 was used as reference strain allowing a comparison with the other strains. Out of the 188 strains tested, 140 strains (∼75%) presented a significantly higher probability of growth under low pH conditions than the O157:H7 strain ATCC 43888, whereas 20 strains (∼11%) showed a significantly lower probability of growth under high pH conditions.

Lysozyme inhibitor conferring bacterial tolerance to invertebrate type lysozyme.

Invertebrate (I-) type lysozymes, like all other known lysozymes, are dedicated to the hydrolysis of peptidoglycan, the major bacterial cell wall polymer, thereby contributing to the innate immune system and/or digestive system of invertebrate organisms. Bacteria on the other hand have developed several protective strategies against lysozymes, including the production of periplasmic and/or membrane-bound lysozyme inhibitors. The latter have until now only been described for chicken (C-) type lysozymes. We here report the discovery, purification, identification and characterization of the first bacterial specific I-type lysozyme inhibitor from Aeromonas hydrophila, which we designate PliI (periplasmic lysozyme inhibitor of the I-type lysozyme). PliI has homologs in several proteobacterial genera and contributes to I-type lysozyme tolerance in A. hydrophila in the presence of an outer membrane permeabilizer. These and previous findings on C-type lysozyme inhibitors suggest that bacterial lysozyme inhibitors may have an important function, for example, in bacteria-host interactions.

Spike synchronization and rate modulation differentially involved in motor cortical function.

It is now commonly accepted that planning and execution of movements are based on distributed processing by neuronal populations in motor cortical areas. It is less clear, though, how these populations organize dynamically to cope with the momentary computational demands. Simultaneously recorded activities of neurons in the primary motor cortex of monkeys during performance of a delayed-pointing task exhibited context-dependent, rapid changes in the patterns of coincident action potentials. Accurate spike synchronization occurred in relation to external events (stimuli, movements) and was commonly accompanied by discharge rate modulations but without precise time locking of the spikes to these external events. Spike synchronization also occurred in relation to purely internal events (stimulus expectancy), where firing rate modulations were distinctly absent. These findings indicate that internally generated synchronization of individual spike discharges may subserve the cortical organization of cognitive motor processes.

Dynamics of ongoing activity: explanation of the large variability in evoked cortical responses.

Evoked activity in the mammalian cortex and the resulting behavioral responses exhibit a large variability to repeated presentations of the same stimulus. This study examined whether the variability can be attributed to ongoing activity. Ongoing and evoked spatiotemporal activity patterns in the cat visual cortex were measured with real-time optical imaging; local field potentials and discharges of single neurons were recorded simultaneously, by electrophysiological techniques. The evoked activity appeared deterministic, and the variability resulted from the dynamics of ongoing activity, presumably reflecting the instantaneous state of cortical networks. In spite of the large variability, evoked responses in single trials could be predicted by linear summation of the deterministic response and the preceding ongoing activity. Ongoing activity must play an important role in cortical function and cannot be ignored in exploration of cognitive processes.

Role of the alternative sigma factor sigma on Staphylococcus aureus resistance to stresses of relevance to food preservation.

AIMS: To examine the role of the alternative general stress sigma factor sigma(B) on the resistance of Staphylococcus aureus to stresses of relevance to food preservation, with special emphasis on emerging technologies such as pulsed electric fields (PEF) and high hydrostatic pressure (HHP). METHODS AND RESULTS: S. aureus strain Newman and its isogenic DeltasigB mutant were grown to exponential and stationary growth phases and its resistance to various stresses was tested. The absence of the sigma(B) factor caused a decrease in the resistance to heat, PEF, HHP, alkali, acid and hydrogen peroxide. In the case of heat, the influence of the sigma(B) factor was particularly important, and decreases in decimal reduction time values of ninefold were observed as a result of its deficiency. The increased thermotolerance of the parental strain as compared with the sigB mutant could be attributed to a better capacity to sustain and repair sublethal damages caused by heat. CONCLUSIONS: sigma(B) factor provides S. aureus cells with resistance to multiple stresses, increasing survival to heat, PEF and HHP treatments. SIGNIFICANCE AND IMPACT OF THE STUDY: Results obtained in this work help in understanding the physiological mechanisms behind cell survival and death in food-processing environments.

Response synchronization in the visual cortex.

Recently, the classic idea of a 'relational' code in the brain received new impetus from the observation of stimulus-dependent synchronizations in the visual cortex. Work over the past two years has focused on documenting the functional dependencies of such synchronizations. We review and classify the experimental findings with respect to separate spatial and temporal schemes. Thus, we distinguish between different signal classes (spikes, continuous signals), and different types of time-locking to the stimulus. These various classes of synchronization phenomena are discussed with regard to their properties, the underlying mechanisms and their possible relevance for visual processing.

Validation of predictive growth models describing superatmospheric oxygen effects on Pseudomonas fluorescens and Listeria innocua on fresh-cut lettuce.

Two microbial growth models predicting the growth of Pseudomonas fluorescens and Listeria innocua at superatmospheric oxygen and carbon dioxide concentrations at 7 degrees C were validated on fresh-cut butterhead lettuce. Cut lettuce was inoculated with the same strain of L. innocua as the in vitro experiments. The P. fluorescens strain was tagged with a gene encoding green fluorescent protein (GFP) in order to distinguish the inoculated strain from contaminating Pseudomonaceae. Also growth of aerobic mesophilic and lactic acid bacteria was monitored during the experiments. The suggested P. fluorescens model was appropriate to predict growth on cut lettuce. L. innocua on the other hand, grew considerably slower under in vivo circumstances than predicted. CO(2) had a growth promoting effect on L. innocua growing on cut lettuce, whereas in vitro an inhibiting effect was observed. Validation parameters are calculated and hypotheses to explain the discrepancy between predicted and observed growth of L. innocua are provided.

Morphological organization of rat hippocampal slice cultures.

Using various histological methods, we investigated the cellular and morphological organization of rat hippocampal slice cultures. Many of the typical features of the hippocampus were retained in vitro over a long period of time. The principal cell types of the hippocampus and dentate gyrus, the pyramidal cells and granule cells, were well preserved and matured in vitro. Nonpyramidal cells and gamma-aminobutyric-acid (GABA) cells were also present in slice cultures and exhibited a strikingly similar dendritic appearance at the light microscopic level. Moreover, GABA-immunoreactive cell bodies and presynaptic terminals could be identified at the electron microscopic level; they expressed typical symmetric synaptic contacts with cell bodies and dendrites. The course of the intrinsic hippocampal fiber pathways--the mossy fibers, Schaffer collaterals, and alveus--was generally retained in vitro. Additional aberrant fiber projections could be identified. Finally, three types of nonneuronal cells could be distinguished on the basis of immunocytochemical methods.

Neural dynamics in cortex-striatum co-cultures--II. Spatiotemporal characteristics of neuronal activity.

Neural dynamics in organotypic cortex-striatum co-cultures grown for three to six weeks under conditions of dopamine deficiency are described. Single neuron activities were recorded intra- and extracellularly, and spatiotemporal spreading of population activity was mapped using voltage-sensitive dyes. The temporal properties of spike firing were characterized by interspike interval histograms, autocorrelation and crosscorrelation. Cortical pyramidal neurons (n = 40) showed irregular firing with a weak tendency to burst or to oscillate. Crosscorrelations revealed strong near-coincident firing and synaptic interactions. Disinhibition was a notable feature in a strongly firing cortical interneuron. Cortical activity spread in the co-culture, thus inducing an overall, homogeneous depolarization in the striatal part. Striatal cells were divided into principal cells and type I and II secondary cells. Principal cells (n = 40) were similar to those reported previously in vivo. Spiking activity ranged from irregular spiking at very low rates to episodic bursting, with an average burst duration of 1 s. Interspike intervals were single-peaked. Intracellular recordings revealed characteristic, long-lasting subthreshold depolarizations ("enabled state") that were shortened by local muscarinic receptor blockade. During prolonged time periods in the "enabled state", locally applied bicuculline induced strong firing in most principal neurons. Striatal secondary type I neurons (n = 25) showed high spiking rates, single- and double-peaked interval histograms and low-threshold, short-lasting stereotyped bursting activity and occasional rhythmic bursting. The firing of these neurons was increased by bicuculline. Crosscorrelations showed synchronization of these cells with principal cell activity. Secondary type II neurons (n = 15) revealed tonic, irregular firing patterns similar to cortical neurons, except with occasional firing in doublet spikes. We conclude that under conditions of dopamine deficiency in corticostriatal co-cultures (i) the cortex induces the "enabled" state and typical bursting mode in striatal principal neurons; (ii) principal neurons are strongly inhibited during the "enabled" state; (iii) muscarinic activity, presumably from tonically active striatal cholinergic interneurons, stabilizes the "enabled" state; (iv) striatal GABAergic interneurons receives synaptic inhibition and take part in synchronized activity among striatal principal cells. Our results favor the view of the striatum as a lateral inhibition network.

Neural dynamics in cortex-striatum co-cultures--I. anatomy and electrophysiology of neuronal cell types.

An in vitro system was established to analyse corticostriatal processing. Cortical and striatal slices taken at postnatal days 0-2 were co-cultured for three to six weeks. The anatomy of the organotypic co-cultures was determined using immunohistochemistry. In the cortex parvalbumin-positive and calbindin-positive cells, which resembled those seen in vivo, had laminar distributions. In the striatum, strongly stained parvalbumin-positive cells resembling striatal GABAergic interneurons and cholinergic interneurons were scattered throughout the tissue. The soma area of these interneuron classes was larger than the average striatal soma area, thus enabling visual selection of cells by class before recording. Cortical neurons with projections to the striatum showed similar morphological features to corticostriatal projection neurons in vivo. No projections from the striatum to the cortex were found. Intracellular recordings were obtained from 94 neurons. These were first classified on the basis of electrophysiological characteristics and the morphologies of cells in each class were reconstructed. Two types of striatal secondary neurons with unique electrophysiological dynamics were identified: GABAergic interneurons (n = 17) and large aspiny, probably cholinergic, interneurons (n = 15). The electrophysiological and morphological characteristics of cortical pyramidal cells (n = 27), cortical interneurons (n = 1), as well as striatal principal neurons (n = 34), were identical to those reported for similar ages in vivo. Organotypic cortex-striatum co-cultures are therefore suitable as an in vitro system in which to analyse corticostriatal processing. The network dynamics, which developed spontaneously in that system, are examined in the companion paper.

'Dynamics of neuronal interactions' cannot be explained by 'neuronal transients'.

In a recent paper, Vaadia et al. demonstrated that patterns of firing correlation between single neurons in the cortex of behaving monkeys can be modified within a fraction of a second. These changes occur in relation to sensory stimuli and behavioral events, and even without modulations of the neurons' firing rates. These findings call for a revision of prevailing models of neural coding that solely rely on single neuron firing rates. In a defense of these models, Friston put forward an alternative explanation, proposing that the observed correlation dynamics emerge solely from co-modulations of the firing rates of each of the neurons, while the strength of their interaction remains constant. To test this possibility we re-examined the data, adopting Friston's 'neuronal transients' model, and the associated equations and procedures. We found that, to explain the dynamic correlation between a pair of neurons, the alternative interpretation requires that each neuron's response to a single stimulus is composed of a relatively large number of independent components, which co-vary with their counterparts in the companion neuron. This large number of components and their shapes lead us to conclude that, although in principle possible, the neuronal transients model: (i) does not provide a simpler explanation of the experimental results; and (ii) cannot explain these results without itself deviating significantly from most rate code models.

Neural dynamics in cortical networks--precision of joint-spiking events.

Electrophysiological studies of cortical function on the basis of multiple single-neuron recordings reveal neuronal interactions which depend on stimulus context and behavioural events. These interactions exhibit dynamics on different time scales, with time constants down to the millisecond range. Mechanisms underlying such dynamic organization of the cortical network were investigated by experimental and theoretical approaches. We review some recent results from these studies, concentrating on the occurrence of precise joint-spiking events in cortical activity, both in physiological and in model neural networks. These findings suggest that a combinatorial neural code, based on rapid associations of groups of neurons co-ordinating their activity at the single spike level, is biologically feasible.

Propagation of synchronous spiking activity in feedforward neural networks.

'Synfire' activity has been proposed as a model for the experimentally observed accurate spike patterns in cortical activity. We investigated the structural and dynamical aspects of this theory. To quantify the degree of synchrony in neural activity, we introduced the concept of 'pulse packets'. This enabled us to derive a novel neural transmission function which was used to assess the role of the single neuron dynamics and to characterize the stability conditions for propagating synfire activity. Thus, we could demonstrate that the cortical network is able to sustain synchronous spiking activity using local feedforward (synfire) connections. This new approach opens the way for a quantitative description of neural network dynamics, and enables us to test the synfire hypothesis on physiological data.

Single-trial estimation of neuronal firing rates: from single-neuron spike trains to population activity.

We present a method to estimate the neuronal firing rate from single-trial spike trains. The method, based on convolution of the spike train with a fixed kernel function, is calibrated by means of simulated spike trains for a representative selection of realistic dynamic rate functions. We derive rules for the optimized use and performance of the kernel method, specifically with respect to an effective choice of the shape and width of the kernel functions. An application of our technique to the on-line, single-trial reconstruction of arm movement trajectories from multiple single-unit spike trains using dynamic population vectors illustrates a possible use of the proposed method.

A low-cost single-board solution for real-time, unsupervised waveform classification of multineuron recordings.

We describe a low-cost single-board system for unsupervised, real-time spike sorting of recordings from a number of neurons on a single microelectrode. The maximum number of spike classes depends on the quality of the recording; it will typically be between 2 and 5. The spike sorter communicates with a conventional microcomputer through a standard serial port (RS232). For typical firing rates as measured in the mammalian central nervous system, this set-up will accommodate up to some 10 parallel spike sorters for as many separate microelectrodes.

Detecting unitary events without discretization of time.

In earlier studies we developed the 'Unitary Events' analysis (Grün S. Unitary Joint-Events in Multiple-Neuron Spiking Activity: Detection, Significance and Interpretation. Reihe Physik, Band 60. Thun, Frankfurt/Main: Verlag Harri Deutsch, 1996.) to detect the presence of conspicuous spike coincidences in multiple single unit recordings and to evaluate their statistical significance. The method enabled us to study the relation between spike synchronization and behavioral events (Riehle A, Grün S, Diesmann M, Aertsen A. Spike synchronization and rate modulation differentially involved in motor cortical function. Science 1997;278:1950-1953.). There is recent experimental evidence that the timing accuracy of coincident spiking events, which might be relevant for higher brain function, may be in the range of 1-5 ms. To detect coincidences on that time scale, we sectioned the observation interval into short disjunct time slices ('bins'). Unitary Events analysis of this discretized process demonstrated that coincident events can indeed be reliably detected. However, the method looses sensitivity for higher temporal jitter of the events constituting the coincidences (Grün S. Unitary Joint-Events in Multiple-Neuron Spiking Activity: Detection, Significance and Interpretation. Reihe Physik, Band 60. Thun, Frankfurt/Main: Verlag Harri Deutsch, 1996.). Here we present a new approach, the 'multiple shift' method (MS), which overcomes the need for binning and treats the data in their (original) high time resolution (typically 1 ms, or better). Technically, coincidences are detected by shifting the spike trains against each other over the range of allowed coincidence width and integrating the number of exact coincidences (on the time resolution of the data) over all shifts. We found that the new method enhances the sensitivity for coincidences with temporal jitter. Both methods are outlined and compared on the basis of their analytical description and their application on simulated data. The performance on experimental data is illustrated.

Evaluation of neuronal connectivity: sensitivity of cross-correlation.

Cross-correlation analysis of separable multi-unit activity is one of the most commonly used methods to investigate connectivity in neural networks. In the course of development of new analysis techniques which go beyond the study of pairs or triplets of neurons, the need arose for a simple yet versatile simulator to generate spike trains from networks of specified structure. The present paper describes such a simulator and presents some examples of its performance as analyzed by cross-correlation. We noted a distinct asymmetry in the sensitivity of cross-correlation for the presence of excitatory vs inhibitory connections. A theoretical analysis is given from which quantitative criteria for detectability were derived. It appears that indeed the sensitivity of cross-correlation for excitation is larger to an order of magnitude than it is for inhibition. Possible consequences of this finding are indicated, and the relation to commonly used methods to measure strength of interaction are discussed.

Rhythmic and patterned neuronal firing in visual cortex.

We present evidence from multi-electrode recordings that the stimulus driven activity in a region of monkey visual cortex displays synchronous, oscillatory behaviour with temporally changing patterns of firing. The spatial-temporal patterns are dependent on cortical layer and on visual stimulus, 'build up and die down' with a period of 70-100 msec, and cannot be factorized into separate spatial and temporal components. This finding supports neural network models which explicitly incorporate spatial-temporal structure, ruling against a purely spatial code for information processing. The new type of analysis presented here is also suitable for direct on-line interactive analysis during multi-electrode recording experiments. Further, it could be modified for use in human EEG or evoked response recordings.

Directional hearing in the grass frog (Rana temporaria L.): I. Mechanical vibrations of tympanic membrane.

The vibration characteristics (amplitude and phase as a function of frequency) of the tympanic membrane in the grass frog were measured using a laser-doppler velocity meter. It was tested to what extent the frog's acoustic system behaves as a pressure gradient receiver. This might clarify how the frog localizes sound. Using a closed sound system the membrane was stimulated at three different entrances: in front of the membrane, at the contralateral ear and from inside the mouth. A combination of these can describe the motion of the membrane under free field conditions. It is found that the sound entrance from inside the mouth will give almost identical vibration characteristics as stimulation in front of the membrane. This can yield a perfect gradient receiver mechanism, when the frog opens its mouth. It is doubted however whether the frog in nature needs to open its mouth for localization of sound. With mouth closed the effectiveness of the gradient receiver will be determined by the transmission characteristics of sound across the tissues of the mouth. The entrance of sound via the contralateral ear is only effective at frequencies between 800 and 1600 Hz. At those frequencies crosstalk between the membranes is however not more than -4 to -8 dB. This is subject to changes in the acoustic properties of the mouth cavity and can possibly be altered by the frog in free nature.