Spatial localization of calcium channels in giant fiber lobe neurons of the squid (Loligo opalescens).

Whole-cell voltage clamp was used to investigate the properties and spatial distribution of fast-deactivating (FD) Ca channels in squid giant fiber lobe (GFL) neurons. Squid FD Ca channels are reversibly blocked by the spider toxin omega-Agatoxin IVA with an IC50 of 240-420 nM with no effect on the kinetics of Ca channel gating. Channels with very similar properties are expressed in both somatic and axonal domains of cultured GFL neurons, but FD Ca channel conductance density is higher in axonal bulbs than in cell bodies at all times in culture. Channels presumably synthesized during culture are preferentially expressed in the growing bulbs, but bulbar Ca conductance density remains constant while Na conductance density increases, suggesting that processes determining the densities of Ca and Na channels in this extrasomatic domain are largely independent. These observations suggest that growing axonal bulbs in cultured GFL neurons are not composed entirely of "axonal" membranes because FD Ca channels are absent from the giant axon in situ but, rather, suggest a potential role for FD Ca channels in mediating neurotransmitter release at the motor terminals of the giant axon.

Spatial expression of two anti-inflammatory mediators, annexin 1 and galectin-1, in nasal polyposis

Background There is renewed interest in the role played by specific counter-regulatory mechanisms to control the inflammatory host response, poorly investigated in human pathology. Here, we monitored the expression of two anti-inflammatory mediators, annexin 1 and galectin-1, and assessed their potential link to glucocorticoids' (GCs) effective control of nasal polyposis (NP).Methods Total patterns of mRNA and protein expression were analysed by quantitative real-time PCR (qPCR) and Western blotting analyses, whereas ultrastructural immunocytochemistry was used for spatial localization and quantification of each mediator, focusing on mast cells, eosinophils and epithelial cells.Results Up-regulation of the annexin 1 gene, and down-regulation of galectin-1 gene, was detected in polypoid tissue compared with nasal mucosa. Patient treatment with betamethasone augmented galectin-1 protein expression in polyps. At the cellular level, control mast cells and eosinophils displayed higher annexin 1 expression, whereas marked galectin-1 immunolabelling was detected in the granule matrix of mast cells. Cells of glandular duct epithelium also displayed expression of both annexin 1 and galectin-1, augmented after treatment.Conclusion Mast cells and epithelial cells appeared to be pivotal cell types involved in the expression of both annexin 1 and galectin-1. It is possible that annexin 1 and galectin-1 could be functionally associated with a specific mechanism in NP and that GC exert at least part of their beneficial effects on the airway mucosa by up-regulating, in a specific cell target fashion, these anti-inflammatory agonists.

Dynamic localization of a cytoplasmic signal transduction response regulator controls morphogenesis during the Caulobacter cell cycle

We present evidence that a bacterial signal transduction cascade that couples morphogenesis with cell cycle progression is regulated by dynamic localization of its components. Previous studies have implicated two histidine kinases, DivJ and PleC, and the response regulator, DivK, in the regulation of morphogenesis in the dimorphic bacterium Caulobacter crescentus. Here, we show that the cytoplasmic response regulator, DivK, exhibits a dynamic, cyclical localization that culminates in asymmetric distribution of DivK within the two cell types that are characteristic of the Caulobacter cell cycle; DivK is dispersed throughout the cytoplasm of the progeny swarmer cell and is localized to the pole of the stalked cell. The membrane-bound DivJ and PleC histidine kinases, which are asymmetrically localized at the opposite poles of the predivisional cell, control the temporal and spatial localization of DivK. DivJ mediates DivK targeting to the poles whereas PleC controls its release from one of the poles at times and places that are consistent with the activities and location of DivJ and PleC in the late predivisional cell. Thus, dynamic changes in subcellular location of multiple components of a signal transduction cascade may constitute a novel mode of prokaryotic regulation to generate and maintain cellular asymmetry.

In vivo examination of membrane protein localization and degradation with green fluorescent protein.

To test the utility of green fluorescent protein (GFP) as an in vivo reporter protein when fused to a membrane domain, we made a fusion protein between yeast hydroxymethylglutaryl-CoA reductase and GFP. Fusion proteins displayed spatial localization and regulated degradation consistent with the native hydroxymethylglutaryl-CoA reductase proteins. Thus, GFP should be useful in the study of both membrane protein localization and protein degradation in vivo.

Modelos da Química Quântica no espaço de momento: diferentes representações de um mesmo sistema

The conventional approach to simple quantum chemistry models is contrasted with that known as momentum representation, where the wavefunctions are momentum dependent. Since the physical interactions are the same, state energies should not change, and whence the energy differences correlating with the real world as spectral lines or bands. We emphasize that one representation is not more fundamental than the other, and the choice is a matter of mathematical convenience. As spatial localization is rooted in our brains, to think in terms of the momentum present us a great mental challenge that can lead to complementary perspectives of a model.

Neuroprotective Effects of Diazepam, Carbamazepine, Phenytoin and Ketamine after Pilocarpine-induced Status Epilepticus

Cell damage and spatial localization deficits are often reported as long-term consequences of pilocarpine-induced status epilepticus. In this study, we investigated the neuroprotective effects of repeated drug administration after long-lasting status epilepticus. Groups of six to eight Wistar rats received microinjections of pilocarpine (2.4 mg/mu l, 1 mu l) in the right dorsal hippocampus to induce a status epilepticus, which was attenuated by thiopental injection (35 mg/kg, i.p.) 3 hrs after onset. Treatments consisted of i.p. administration of diazepam, ketamine, carbamazepine, or phenytoin at 4, 28, 52, and 76 hr after the onset of status epilepticus. Two days after the treatments, rats were tested in the Morris water maze and 1 week after the cognitive tests, their brains were submitted to histology to perform haematoxylin and eosin staining and glial fibrillary acidic protein (GFAP) immunofluorescence detection. Post-status epilepticus rats exhibited extensive gliosis and cell loss in the hippocampal CA1, CA3 (70% cell loss for both areas) and dentate gyrus (60%). Administration of all drugs reduced cell loss in the hippocampus, with best effects observed in brains slices of diazepam-treated animals, which showed less than 30% of loss in the three areas and decreased GFAP immunolabelling. Treatments improved spatial navigation during training trials and probe trial, with exception of ketamine. Interestingly, in the probe trial, only diazepam-treated animals showed preference for the goal quadrant. Our data point to significant neuroprotective effects of repeated administration of diazepam against status epilepticus-induced cell damage and cognitive disturbances.

Direct visualization of Ras proteins in spatially distinct cell surface microdomains

Localization of signaling complexes to specific micro-domains coordinates signal transduction at the plasma membrane. Using immunogold electron microscopy of plasma membrane sheets coupled with spatial point pattern analysis, we have visualized morphologically featureless microdomains including lipid rafts, in situ and at high resolution. We find that an inner-plasma membrane lipid raft marker displays cholesterol-dependent clustering in microdomains with a mean diameter of 44 nm that occupy 35% of the cell surface. Cross-linking an outer-leaflet raft protein results in the redistribution of inner leaflet rafts, but they retain their modular structure. Analysis of Ras microlocalization shows that inactive H-ras is distributed between lipid rafts and a cholesterol-independent micro-domain. Conversely, activated H-ras and K-ras reside predominantly in nonoverlapping, cholesterol-independent microdomains. Galectin-1 stabilizes the association of activated H-ras with these nonraft microdomains, whereas K-ras clustering is supported by farnesylation, but not geranylgeranylation. These results illustrate that the inner plasma membrane comprises a complex mosaic of discrete microdomains. Differential spatial localization within this framework can likely account for the distinct signal outputs from the highly homologous Ras proteins.

Advanced instrumentation for superheated liquid detectors in dark matter searches

Dissertação para obtenção do Grau de Doutor em Engenharia Física

Epidemiological aspects of human and canine visceral leishmaniasis in Montes Claros, State of Minas Gerais, Brazil, between 2007 and 2009

INTRODUCTION: Visceral leishmaniasis (VL) is an expanding zoonosis in Brazil and is becoming urbanized in several Brazilian regions. This study aims to describe the epidemiological features of human and canine VL in the municipality of Montes Claros, State of Minas Gerais, by focusing on their spatial distribution. METHODS: Data concerning human cases and reactive dogs for VL from 2007 to 2009 were obtained from the Information System for Disease Notification (SINAN) and from reports of the local Centro de Controle de Zoonoses (CCZ), respectively. The addresses of human and canine cases have been georeferenced and localized in thematic maps, allowing their spatial visualization as well as the identification of areas at risk of VL transmission. RESULTS: Ninety-five cases of human VL were reported in the period. The 0-9-year-old age group (48.4%) was the most affected, within which the majority consisted of male patients (64%). Of the samples collected for the canine serological survey, 2,919 (6.3%) were reactive to VL. The spatial localization of these cases shows that the disease was scattered in the urban area of the municipality. Areas showing a higher dissemination risk were concentrated in the central, northwestern, and southern regions of the city. CONCLUSIONS: Identifying the areas most at risk in urban Montes Claros may help guide actions toward local epidemiological vigilance and control.

Evolution in heterogeneous populations: from migration models to fixation probabilities.

Although dispersal is recognized as a key issue in several fields of population biology (such as behavioral ecology, population genetics, metapopulation dynamics or evolutionary modeling), these disciplines focus on different aspects of the concept and often make different implicit assumptions regarding migration models. Using simulations, we investigate how such assumptions translate into effective gene flow and fixation probability of selected alleles. Assumptions regarding migration type (e.g. source-sink, resident pre-emption, or balanced dispersal) and patterns (e.g. stepping-stone versus island dispersal) have large impacts when demes differ in sizes or selective pressures. The effects of fragmentation, as well as the spatial localization of newly arising mutations, also strongly depend on migration type and patterns. Migration rate also matters: depending on the migration type, fixation probabilities at an intermediate migration rate may lie outside the range defined by the low- and high-migration limits when demes differ in sizes. Given the extreme sensitivity of fixation probability to characteristics of dispersal, we underline the importance of making explicit (and documenting empirically) the crucial ecological/ behavioral assumptions underlying migration models.

Nuclear pairing: surface or bulk?

We analyze how the spatial localization properties of pairing correlations are changing in a major neutron shell of heavy nuclei. It is shown that the radial distribution of the pairing density depends strongly on whether the chemical potential is close to a low or a high angular momentum level and has little sensitivity to whether the pairing force acts at the surface or in the bulk. The pairing density averaged over one major shell is, however, rather flat, exhibiting little dependence on the pairing force. Hartree-Fock-Bogoliubov calculations for the isotopic chain 100-132Sn are presented for demonstration purposes.

Modelos da Química Quântica no espaço de momento: diferentes representações de um mesmo sistema

The conventional approach to simple quantum chemistry models is contrasted with that known as momentum representation, where the wavefunctions are momentum dependent. Since the physical interactions are the same, state energies should not change, and whence the energy differences correlating with the real world as spectral lines or bands. We emphasize that one representation is not more fundamental than the other, and the choice is a matter of mathematical convenience. As spatial localization is rooted in our brains, to think in terms of the momentum present us a great mental challenge that can lead to complementary perspectives of a model.

Vidéosurveillance intelligente pour la détection de chutes chez les personnes âgées

Les pays industrialisés comme le Canada doivent faire face au vieillissement de leur population. En particulier, la majorité des personnes âgées, vivant à domicile et souvent seules, font face à des situations à risques telles que des chutes. Dans ce contexte, la vidéosurveillance est une solution innovante qui peut leur permettre de vivre normalement dans un environnement sécurisé. L’idée serait de placer un réseau de caméras dans l’appartement de la personne pour détecter automatiquement une chute. En cas de problème, un message pourrait être envoyé suivant l’urgence aux secours ou à la famille via une connexion internet sécurisée. Pour un système bas coût, nous avons limité le nombre de caméras à une seule par pièce ce qui nous a poussé à explorer les méthodes monoculaires de détection de chutes. Nous avons d’abord exploré le problème d’un point de vue 2D (image) en nous intéressant aux changements importants de la silhouette de la personne lors d’une chute. Les données d’activités normales d’une personne âgée ont été modélisées par un mélange de gaussiennes nous permettant de détecter tout événement anormal. Notre méthode a été validée à l’aide d’une vidéothèque de chutes simulées et d’activités normales réalistes. Cependant, une information 3D telle que la localisation de la personne par rapport à son environnement peut être très intéressante pour un système d’analyse de comportement. Bien qu’il soit préférable d’utiliser un système multi-caméras pour obtenir une information 3D, nous avons prouvé qu’avec une seule caméra calibrée, il était possible de localiser une personne dans son environnement grâce à sa tête. Concrêtement, la tête de la personne, modélisée par une ellipsoide, est suivie dans la séquence d’images à l’aide d’un filtre à particules. La précision de la localisation 3D de la tête a été évaluée avec une bibliothèque de séquence vidéos contenant les vraies localisations 3D obtenues par un système de capture de mouvement (Motion Capture). Un exemple d’application utilisant la trajectoire 3D de la tête est proposée dans le cadre de la détection de chutes. En conclusion, un système de vidéosurveillance pour la détection de chutes avec une seule caméra par pièce est parfaitement envisageable. Pour réduire au maximum les risques de fausses alarmes, une méthode hybride combinant des informations 2D et 3D pourrait être envisagée.

Adaptation aux changements induits aux indices spectraux de l’audition spatiale chez l’humain.

Dans le cas de perte auditive, la localisation spatiale est amoindrie et vient entraver la compréhension de la parole et ce, malgré le port de prothèses auditives. La présente étude modifie la forme de l’oreille externe d’individus à l’aide de silicone afin d’induire des changements aux indices spectraux (HRTFs), similaires à ceux causés par des prothèses auditives, et d’explorer les mécanismes perceptifs (visuel, spectral, ou tactile) permettant d’alterner d’un nouvel ensemble à l’ensemble originel de HRTFs une fois les prothèses enlevées. Les résultats démontrent que les participants s’adaptent aux nouveaux HRTFs à l’intérieur de quatre séances d’entraînement. Dès le retrait des prothèses, les participants reviennent à leur performance originale. Il n’est pas possible de conclure avec les données présentes si le changement d’un ensemble de HRTFs à un autre est influencé par un des mécanismes de rétroaction perceptuelle étudié. L’adaptation aux prothèses perdure jusqu’à quatre semaines après leur retrait.

A Study of Morphological Operators with Applications

The present work deals with the A study of morphological opertors with applications. Morphology is now a.necessary tool for engineers involved with imaging applications. Morphological operations have been viewed as filters the properties of which have been well studied (Heijmans, 1994). Another well-known class of non-linear filters is the class of rank order filters (Pitas and Venetsanopoulos, 1990). Soft morphological filters are a combination of morphological and weighted rank order filters (Koskinen, et al., 1991, Kuosmanen and Astola, 1995). They have been introduced to improve the behaviour of traditional morphological filters in noisy environments. The idea was to slightly relax the typical morphological definitions in such a way that a degree of robustness is achieved, while most of the desirable properties of typical morphological operations are maintained. Soft morphological filters are less sensitive to additive noise and to small variations in object shape than typical morphological filters. They can remove positive and negative impulse noise, preserving at the same time small details in images. Currently, Mathematical Morphology allows processing images to enhance fuzzy areas, segment objects, detect edges and analyze structures. The techniques developed for binary images are a major step forward in the application of this theory to gray level images. One of these techniques is based on fuzzy logic and on the theory of fuzzy sets.Fuzzy sets have proved to be strongly advantageous when representing in accuracies, not only regarding the spatial localization of objects in an image but also the membership of a certain pixel to a given class. Such inaccuracies are inherent to real images either because of the presence of indefinite limits between the structures or objects to be segmented within the image due to noisy acquisitions or directly because they are inherent to the image formation methods.