Modeling the slow wave shapes of spreading depression in a rat cortex: a methodology for seeking physiological parameters.

Spreading depression (SD) consists of a transient significant suppression of the spontaneous neural electrical activity that spreads slowly across regions of the gray matter in a wave form. Nowadays, this phenomenon is being studied by means of mathematical and computational models to reproduce the main characteristics of SD. Given the high number of parameters and their unknown ranges of variation, the setting of parameters for current SD models is usually a hard task that must be addressed in order to make such models reproduce real data. In this paper, we present a 1-D model which is able to reproduce the most important characteristics of SD waves observed in laboratory experiments: the slow extracellular potential shift and extracellular ionic concentration variations regarding speed, shape, and amplitude. Such a reproduction is possible due to a methodology that we introduced to set the parameters of the SD models. The methodology allows the impact of each parameter on the results produced by the model and the range of parameters for which the model displays plausible behavior to be determined. The methodology also helps to identify features that the model cannot produce and it gives insights about what parts of the model should be modified to improve its capacities through the identification of parameters involved with each behavior.

Can the fractal dimension be applied for the early diagnosis of non-proliferative diabetic retinopathy?

The fractal dimension has been employed as a useful parameter in the diagnosis of retinal disease. Avakian et al. (Curr Eye Res 2002; 24: 274-280), comparing the vascular pattern of normal patients with mild to moderate non-proliferative diabetic retinopathy (NPDR), found a significant difference between them only in the macular region. This significant difference in the box-counting fractal dimension of the macular region between normal and mild NPDR patients has been proposed as a method of precocious diagnosis of NPDR. The aim of the present study was to determine if fractal dimensions can really be used as a parameter for the early diagnosis of NPDR. Box-counting and information fractal dimensions were used to parameterize the vascular pattern of the human retina. The two methods were applied to the whole retina and to nine anatomical regions of the retina in 5 individuals with mild NPDR and in 28 diabetic but opthalmically normal individuals (controls), with age between 31 and 86 years. All images of retina were obtained from the Digital Retinal Images for Vessel Extraction (DRIVE) database. The results showed that the fractal dimension parameter was not sensitive enough to be of use for an early diagnosis of NPDR.

Can the fractal dimension be applied for the early diagnosis of non-proliferative diabetic retinopathy?

The fractal dimension has been employed as a useful parameter in the diagnosis of retinal disease. Avakian et al. (Curr Eye Res 2002; 24: 274-280), comparing the vascular pattern of normal patients with mild to moderate non-proliferative diabetic retinopathy (NPDR), found a significant difference between them only in the macular region. This significant difference in the box-counting fractal dimension of the macular region between normal and mild NPDR patients has been proposed as a method of precocious diagnosis of NPDR. The aim of the present study was to determine if fractal dimensions can really be used as a parameter for the early diagnosis of NPDR. Box-counting and information fractal dimensions were used to parameterize the vascular pattern of the human retina. The two methods were applied to the whole retina and to nine anatomical regions of the retina in 5 individuals with mild NPDR and in 28 diabetic but opthalmically normal individuals (controls), with age between 31 and 86 years. All images of retina were obtained from the Digital Retinal Images for Vessel Extraction (DRIVE) database. The results showed that the fractal dimension parameter was not sensitive enough to be of use for an early diagnosis of NPDR.

Fractal analysis of retinal vessel patterns in ophthalmically normal dogs.

OBJECTIVE: To study the applicability of the fractal dimension as a parameter for describing retinal vessel patterns in ophthalmically normal dogs. PROCEDURES: The following strategy was adopted: (i) development of an experimental procedure to obtain digitalized photographs of the fundus; (ii) development of software to segment retinal vessel images and calculate the box-counting and radius of gyration fractal dimensions of the retinal vessels and diffusion-limited aggregation (DLA), a process with similar characteristics to retinal vessel morphology, and (iii) establishment of a standard curve for the fractal dimensions of segmented vessels. RESULTS: Digitalized photographs of the fundus showed an adequate contrast between the vessels and the rest of the fundus for numerical analysis. The software developed produced a binary image of the retinal vessels permitting calculation of the fractal dimension. The mean values of the fractal dimensions calculated by the methods of box-counting and radius of gyration for the DLA were significantly different (t = -40.33, P approximately 0). The radius of gyration method was found to be more suitable for documenting the dimension of the DLA and, consequently, of the dog's retinal vessels. CONCLUSION: This methodology may be useful to differentiate between normal and pathologic states of canine retinal vascularization.

Neuronal nicotinic acetylcholine receptor activation modulates gamma-aminobutyric acid release from CA1 neurons of rat hippocampal slices.

In the present study we investigated electrophysiologically the nicotinic responses of pyramidal neurons and interneurons visualized by infrared-assisted videomicroscopy and fluorescence in the CA1 field of hippocampal slices obtained from 8- to 24-day-old rats. Application of nicotinic agonists to CA1 neurons evoked at least four types of nicotinic responses. Of major interest was the ability of these agonists to induce the release of gamma-aminobutyric acid (GABA) from interneurons. Slowly decaying ACh whole-cell currents and GABA-mediated postsynaptic currents could be recorded from pyramidal neurons and interneurons, whereas fast-decaying nicotinic currents and fast current transients were recorded only from interneurons. Nicotinic responses were sensitive to blockade by d-tubocurarine (10 microM), which indicated that they were mediated by nicotinic acetylcholine receptors (nAChRs). The slowly decaying currents, the postsynaptic currents and the fast current transients were insensitive to blockade by the alpha-7 nAChR-specific antagonist methyllycaconitine (up to 1 microM) or alpha-bungarotoxin (100 nM). On the other hand, the slowly decaying nicotinic currents recorded from the interneurons were blocked by the alpha4beta2 nAChR-specific antagonist dihydro-beta-erythroidine, and the fast-desensitizing nicotinic currents were evoked by the alpha-7 nAChR-specific agonist choline. In experimental conditions similar to those used to record nicotinic responses from neurons in slice (i. e., in the absence of tetrodotoxin), we observed that nicotinic agonists can also induce the release of GABA from hippocampal neurons in culture. In summary, these results provide direct evidence for more than one subtype of functional nAChR in CA1 neurons and suggest that activation of nAChRs present in GABAergic interneurons can evoke inhibitory activity in CA1 pyramidal neurons, thereby modulating processing of information in the hippocampus.

Ligand-gated ion channels in acutely dissociated rat hippocampal neurons with long dendrites.

A technique for dissociation of hippocampi of 3-25-day-old rats is described by which pyramidal and bipolar neurons with many long (up to 200 microns) dendrites can be obtained. Dissociation of CA1 neurons was achieved by mechanical means, in the absence of Ca2+, and without the use of proteolytic enzymes. The functional properties of the dissociated neurons were assessed using the whole-cell patch-clamp technique. Whole-cell currents were elicited by U-tube application of the agonists N-methyl-D-aspartate (NMDA), gamma-aminobutyric acid (GABA), and acetylcholine (ACh), and spontaneous miniature currents were also observed in these neurons. ACh-elicited currents were blocked by methyllycaconitine (MLA, 1 nM) and Pb2+ (0.1-10 microM). These results establish acutely dissociated neurons as a simple and reliable preparation for the study of the pharmacology, kinetics and subcellular distribution of ligand-gated ion channels.

Antigen F1 from Yersinia pestis forms aqueous channels in lipid bilayer membranes.

Antigen F1 is a protein of 17 kDa produced by Yersinia pestis when it is cultured at 37 degrees C. When incorporated into planar lipid bilayer membranes this protein induces fluctuations on membrane conductance typical of the formation of ionic channels. These fluctuations reveal two distinct unitary conductance sizes, one in the range of 800 to 1400 pS and the other in the range of 140 to 600 pS. Zero current potential measurements in the presence of a salt gradient show that the channel is not significantly ion selective. The reversal potential measured in the presence of 0.5 M KCl on the cis side and 0.1 M KCl on the trans side was 3.58 +/- 3.98 mV (N = 7). The non-selectivity of the channel, in addition to its large conductance, suggests that it forms large aqueous pores. The present results, taken together with other data showing that antigen F1 inhibits the activity of phagocytic cells, suggest that antigen F1 acts by forming aqueous pores in the membrane of these target cells.

Antigen F1 from yersinia pestis forms aqueous channels in lipid bilayer membranes

Antigen F1 is a protein of 17 kDa produced by Yersinia pestis it is cultured at 37 grade C. When incorporated into planar lipid bilayer membranes this protein induces fluctuations on membrane conductance typical of the formation of ionic channels. These fluctuations reveal two distinct unitary conductance sizes, one in the range of 800 to 1400 pS and the other in the range of 140 to 600 pS. Zero current potential measaurements in the presence of a salt gradient show that the channell is not significantly ion selective. The reversal potential measured in the presence of 0.5 MKCl on the cis side and 0.1 MKCl on the trans side was 3.58 ñ 3.98 mV (N=7). The non-selectivity of the channel, in addition to its large conductance, suggests that it forms large aqueous pores. The present results, taken together with other data showing that nantigen F1 inhibits the activity of phagocytic cells, suggest that antigen F1 acts by forming aqueous pores in the membrane of these target cells