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The effect of euplotin C -- a lipophilic bioactive metabolite produced by the ciliate Euplotes crassus -- on the kinetics of both phagocytosis of latex particles and fluid-phase uptake of dextran, was studied in the single-cell ciliate Paramecium primaurelia. The inhibition of food vacuole formation was concentration- and time-dependent (p

GABAB receptors modulate swimming behavior in Paramecium by inhibiting dihydropyridine-sensitive Ca2+ channels via G-proteins. Prolonged occupancy of GABAB receptors by baclofen results in a decrease in GABAB receptor functions. Since changes in the number of cell-surface GABAA receptors have been postulated to be of importance in modulating inhibitory synaptic transmission in neurons, we have studied the cell-surface expression and maintenance of GABAB receptors in P. primaurelia. In this study, we use immunostaining in electron and confocal microscopy to demonstrate that constitutive internalization of GABAB receptors in P. primaurelia is mediated by clathrin-dependent and -independent endocytosis. Indeed, GABAB receptors colocalize with the adaptin complex AP2, which is implicated in the selective recruitment of integral membrane proteins to clathrin-coated vesicles, and with caveolin 1, which is associated with uncoated membrane invaginations. Furthermore, when endocytosis is blocked with hypertonic medium, cytosol acidification, filipin or with a peptide that disrupts the association between amphiphysin and dynamin, the effect of baclofen on swimming is increased. These results suggest that GABAB receptor endocytosis into clathrin-coated and -uncoated vesicles represents an important mechanism in the modulation of swimming behavior in Paramecium.

The presence of opioid, beta-adrenergic and cholinergic receptors has been demonstrated in ciliated protozoa, but little is known about gamma-aminobutyric acid (GABA) receptors. In this study we have analyzed the distribution of GABA(A)-type receptor subunits in Paramecium. Confocal laser microscopy using antibodies specific for alpha(1)-, alpha(2)-, alpha(3)-, alpha(6)-, beta(2/3)-, gamma(2)-, epsilon-, lambda-, and theta-subunits showed that most receptors are aggregated in clusters and are distributed both on cell surface and in the cytoplasm. The intensity of labelling of the alpha(6)-, beta(2/3)- and gamma(2)-subunits was more intense than the alpha(1)-, epsilon-, and theta-subunits, suggesting that the former are present in higher concentrations than the latter.

In Paramecium, internal Ca(2+) concentration increase coupled to membrane depolarization induces a reversal in the direction of ciliary beating and, consequently, a reversal in swimming direction. The ciliary reversal (CR) duration is correlated to Ca(2+) influx, and the addition of drugs that block the Ca(2+) current leads to a reduction in the backward swimming duration. In this study we have examined the possible function of GABA(B) receptors in P. primaurelia swimming control. The presence of GABA(B) immunoanalogue in Paramecium was evidenced using SDS-PAGE, Western blotting, and confocal laser scanning microscopy. By applying the specific GABA(B) receptor agonist baclofen, a dose-dependent inhibition of the membrane depolarization-induced CR duration was observed. This inhibition was antagonized by phaclofen, persisted when K(+) channel blockers were applied, and disappeared after treatment with nifedipine and verapamil. Moreover, the action of baclofen on depolarization-induced CR was suppressed by treatment with pertussis toxin. Therefore, these experiments suggest that baclofen modulates CR by a G protein (G(0) or G(1)) mediated inhibition of dihydropyridine-sensible calcium channels. Finally, synthesis and release of GABA in the environment by Paramecium have been demonstrated by HPLC. Possible correlations between GABA(B) receptor activation and the regulation of intracellular Ca(2+) levels are discussed.

Starting from MR and CT images for a given patient, a new single image representation of all parameters has been generated by using false-color techniques in a standard UNIX and X-11 environment. A transformation linking together the MR, CT parameters and the RGB (red, green, blue) color components has been used. Moreover an unsupervised segmentation method based on a number of neural and fuzzy models may directly produce segmented image volumes. Each image of the various sequences has been interactively displayed by using a specifically designed application. The resulting images have been displayed on a stereo monitor allowing the three-dimensional rendering of visual data through LCD shuttered glasses. Moreover, a 3-D control system based on low frequency magnetic fields has been used, while a bandheld Polhemus stylus could be used as an electronic knife for dissecting the 3-D data set and for defining flaps and grafts. Bone or soft-tissue contour can be analyzed, and sections can be removed from the model to allow a view of the underlying structures. Flaps and grafts obtained utilizing the above-reported techniques can be fitted exactly, without repeated removal and recarving. Nuances of depth, tapering, and arc are carved directly into the bone, while chances of asymmetry are markedly diminished. In this way, moreover, anesthetic times are reduced by more efficient utilization of operative time, which usually offsets the increased cost of imaging.

Total 3-D reconstruction of the tumor size, shape, and relations with surrounding structures using CT, MRI, sonography, and angiography images can make simulated radical resection of soft-tissue sarcomas possible, thus sparing normal tissues. With our approach, starting from three MR images for a given patient, a new single image representation of all three parameters is generated by using two different techniques on a workstation in a standard UNIX and X-11 environment. The first one is a transformation linking together the MR parameters and the RGB (red, green, blue) color components. The second one is an unsupervised segmentation method based on a number of neural and fuzzy models. We can dinamically render and update a stereo display using field sequential presentation of left and right eye views on the monitor, with Cristal Eyes LCD shutter eyewear (StereoGraphics Inc., San Rafael, CA) to view it. As 3D locating tool, a 3D locating control system based on low-frequency magnetic fields (Polhemus Fastrak) has been chosen. Simulations of soft-tissues excisions may be performed in this interactive environment with augmented-reality modalities. All this, in our experience, has greatly facilitated the simulation of soft-tissue sarcoma excisions.

We have developed two telecare applications based on mobile telephony (WAP) and WEB. The first can be used to request Basic Life Support (BLS) guidelines any time by using a WAP device and to teach people and non-professionals involved in health care emergency situations. The second is a WEB-WAP based tool for medical data retrieval and at-home health care monitoring of chronically ill patients with congestive heart failure (CHF) or diabetes. Medical education content related to these diseases is available on the WEB and on the WAP device. The WAP application uses the features found in the last generation of mobile phones such as better multimedia information presentations, better interactivity capabilities, and enhanced ease of use. Based on these two applications, a promising platform is offered for developing applications in health care, home care, medical monitoring and health education ensuring continuity of care. In the paper we present the preliminary results of a pilot test at Thessaloniki University (Greece) where the WEB-WAP based tool is used to monitor patients with diabetes or CHF.

The aim of this study has been the biochemical demonstration of the presence of gamma-aminobutyric acid (GABA) in the Mediterranean sea fan Eunicella cavolini by means of high-performance liquid chromatography, and the description of the distribution pattern of GABA and its related molecules, glutamic acid decarboxylase (GAD), vesicular GABA transporter (VGAT) and one of the GABA receptors (GABA(B) R) by immunohistochemical methods. The interrelationships of GABA, GAD and GABA receptor immunoreactivity have been established by using double-immunohistochemical methods and confocal microscopy. The immunodetection of monoclonal and/or polyclonal antibodies has revealed GABA immunoreactivity throughout the polyp tissue, both in neuronal and non-neuronal elements. GAD immunoreactivity has been mostly localized in the neuronal compartment, contacting epithelial and muscular elements. GABA(B) R immunoreactivity appears particularly intense in the nematocytes and in the oocyte envelope; its presence in GAD-immunoreactive neurons in the tentacles suggests an autocrine type of regulation. Western blot analysis has confirmed that a GABA(B) R, with a molecular weight of 142 kDa, similar to that of rat brain, is present in E. cavolini polyp tissue. The identification of the sites of the synthesis, vesicular transport, storage and reception of GABA strongly suggests the presence of an almost complete set of GABA-related molecules for the functioning of the GABAergic system in this simple nervous system. The distribution of these different immunoreactivities has allowed us to hypothesize GABA involvement in nematocyst discharge, in body wall and enteric muscular contraction, in neuronal integration and in male gametocyte differentiation.