Role of serotonin receptors in panic-like behavior induced by nitric oxide in the rat dorsolateral periaqueductal gray: effects of chronic clomipramine treatment

Local administration of serotonin (5-HT) receptor agonists inhibits panic-like reactions induced by electrical stimulation of the rat dorsolateral periaqueductal grey (dlPAG). This anti-aversive effect is enhanced by chronic treatment with anti-panic drugs such as clomipramine. Since nitric oxide (NO) may mediate panic-like behavior in the dlPAG, we tested the hypothesis that chronic clomipramine treatment would also potentiate the effects of locally injected 5-HT-receptor agonists on panic-like reactions induced by intra-dlPAG injection of an NO-donor (SIN). After 21 days of daily i.p. injections of saline or clomipramine (10 mg/kg) male Wistar rats received local injections of saline, the 5-HT(1A)-receptor agonist 8-OH-DPAT (8 nmol) or the 5-HT2A/2C-receptor agonist DOI (16 nmol) followed by saline or SIN (150 nmol). NO-induced panic-like reactions were inhibited by DOI, but not by 8-OH-DPAT. Chronic clomipramine did not modify these effects but tended to produce anti-aversive effect by itself. In chronically clomipramine treated animals 8-OH-DPAT potentiated NO-induced panic-like reactions. The results indicate that the panic-like effects of NO in the dlPAG may be attenuated by 5-HT2A/2C-, but not by 5-HT1A-receptors. The anti-aversive effect of DOI is not modified by chronic clomipramine treatment.     

Intracellular interference of infectious bursal disease virus

A search for dominant-negative mutant polypeptides hampering infectious bursal disease virus (IBDV) replication has been undertaken. We have found that expression of a mutant version of the VP3 structural polypeptide known as VP3/M3, partially lacking the domain responsible for the interaction with the virus-encoded RNA polymerase, efficiently interferes with the IBDV replication cycle. Transformed cells stably expressing VP3/M3 show a significant reduction (up to 96%) in their ability to support IBDV growth. Our findings provide a new tool for the characterization of the IBDV replication cycle and might facilitate the generation of genetically modified chicken lines with a reduced susceptibility to IBDV infection.     

An experimental approach to the fundamental principles of hemodynamics

An experimental model has been developed to give students hands-on experience with the fundamental laws of hemodynamics. The proposed experimental setup is of simple construction but permits the precise measurements of physical variables involved in the experience. The model consists in a series of experiments where different basic phenomena are quantitatively investigated, such as the pressure drop in a long straight vessel and in an obstructed vessel, the transition from laminar to turbulent flow, the association of vessels in vascular networks, or the generation of a critical stenosis. Through these experiments, students acquire a direct appreciation of the importance of the parameters involved in the relationship between pressure and flow rate, thus facilitating the comprehension of more complex problems in hemodynamics.     

Regulation of membrane trafficking by a novel Cdc42-related protein in Caenorhabditis elegans epithelial cells

Rho GTPases are mainly known for their implication in cytoskeleton remodeling. They have also been recently shown to regulate various aspects of membrane trafficking. Here, we report the identification and the characterization of a novel Caenorhabditis elegans Cdc42-related protein, CRP-1, that shows atypical enzymatic characteristics in vitro. Expression in mouse fibroblasts revealed that, in contrast with CDC-42, CRP-1 was unable to reorganize the actin cytoskeleton and mainly localized to trans-Golgi network and recycling endosomes. This subcellular localization, as well as its expression profile restricted to a subset of epithelial-like cells in C. elegans, suggested a potential function for this protein in polarized membrane trafficking. Consistent with this hypothesis, alteration of CRP-1 expression affected the apical trafficking of CHE-14 in vulval and rectal epithelial cells and sphingolipids (C(6)-NBD-ceramide) uptake and/or trafficking in intestinal cells. However, it did not affect basolateral trafficking of myotactin in the pharynx and the targeting of IFB-2 and AJM-1, two cytosolic apical markers of intestine epithelial cells. Hence, our data demonstrate a function for CRP-1 in the regulation of membrane trafficking in a subset of cells with epithelial characteristics.     

Synaptic clusters of MHC class II molecules induced on DCs by adhesion molecule-mediated initial T-cell scanning

Initial adhesive contacts between T lymphocytes and dendritic cells (DCs) facilitate recognition of peptide-MHC complexes by the TCR. In this report, we studied the dynamic behavior of adhesion and Ag receptors on DCs during initial contacts with T-cells. Adhesion molecules LFA-1- and ICAM-1,3-GFP as well as MHC class II-GFP molecules were very rapidly concentrated at the DC contact area. Binding of ICAM-3, and ICAM-1 to a lesser extent, to LFA-1 expressed by mature but not immature DC, induced MHC-II clustering into the immune synapse. Also, ICAM-3 binding to DC induced the activation of the Vav1-Rac1 axis, a regulatory pathway involved in actin cytoskeleton reorganization, which was essential for MHC-II clustering on DCs. Our results support a model in which ICAM-mediated MHC-II clustering on DC constitutes a priming mechanism to enhance antigen presentation to T-cells.     

Oligomycin strengthens the effect of cyclosporin A on mitochondrial permeability transition by inducing phosphate uptake

The purpose of this work was to assess the effect of oligomycin on the mitochondrial membrane permeability transition. The antibiotic was found to strengthen cyclosporin A (CSA)-induced protection of non-specific permeability, which is triggered by a matrix Ca2+ load in the absence of ADP. Oligomycin also reinforced the protective effect of CSA on carboxyatractyloside-induced pore opening in the absence of ADP, but failed to do so in mitochondria incubated under anaerobic conditions or after addition of CCCP. Analyzing the efflux of matrix Ca2+, we found that mitochondrial swelling and the collapse of the transmembrane electric gradient coincided with membrane leakage. The effects of the antibiotic were observed in phosphate-containing media but not in the presence of acetate. Furthermore, N-ethylmaleimide hindered the protective effect of oligomycin-CSA. In addition, the matrix phosphate concentration increased concurrently with a diminution in the matrix-free fraction of Ca2+. We concluded that oligomycin increases phosphate uptake by stimulating the phosphate-/OH- exchange reaction.     

Filtering of kinematic signals using the Hodrick-Prescott filter

The use of the Hodrick-Prescott (HP) filter is presented as an alternative to the traditional digital filtering and spline smoothing methods currently used in biomechanics. In econometrics, HP filtering is a standard tool used to decompose a macroeconomic time series into a nonstationary trend component and a stationary residual component. The use of the HP filter in the present work is based on reasonable assumptions about the jerk and noise components of the raw displacement signal. Its applicability was tested on 4 kinematic signals with different characteristics. Two are well known signals taken from the literature on biomechanical signal filtering, and the other two were acquired with our own motion capture system. The criterion for the selection of cutoff frequency was based on the power spectral density of the raw displacement signals. The results showed the technique to be well suited to filtering biomechanical displacement signals in order to obtain accurate higher derivatives in a simple and systematic way. Namely, the HP filter and the generalized cross-validated quintic spline (GCVSPL) produce similar RMS errors on the first (0.1063 vs. 0.1024 m/s2) and second (23.76 vs. 23.24 rad/s2) signals. The HP filter performs slightly better than GCVSPL on the third (0.209 vs. 0.236 m/s2) and fourth (1.596 vs. 2.315 m/s2) signals.     

Adoption of shelter dogs in a Brazilian community: assessing the caretaker profile

A survey in Ibiuna, Sao Paulo, Brazil, of caregivers (owners) who adopted shelter dogs assessed length of ownership, proportion of male and female dogs adopted, and owners' characteristics. It addressed breeding, neutering, vaccination, and veterinary care. It used no testing to provide a good “match ”between dog and future owner. Of adopted dogs, 58% were male. Only 36% of owners were located. Mean ownership length was 14.8 months (95% confidence interval = 12.4 to 17.2 months), estimated through a survival analysis method. Of adopted dogs, 40.9% lived with their owners; 34.9% had died (some had lived on the streets); 15.0% were donated; 4.3% ran away; 3.2% were returned to the city shelter. Of interviewees, 57% reported no difficulties with the adoption; 23.1% cited the animal's illness and death as the main difficulty. For contraception, 87 owners (46.7%) chained dogs to prevent contact with other animals; 56.5% were against neutering. Reasons given were compassion (58.1%), unnecessary procedure (11.4%), cost (9.5%), and behavior change (4.8%). This research motivated a design for Ibiuna shelter dog adoption to improve the proportion of successful adoptions.     

An integrated tool for microarray data clustering and cluster validity assessment

SUMMARY: In this paper we present a data mining system, which allows the application of different clustering and cluster validity algorithms for DNA microarray data. This tool may improve the quality of the data analysis results, and may support the prediction of the number of relevant clusters in the microarray datasets. This systematic evaluation approach may significantly aid genome expression analyses for knowledge discovery applications. The developed software system may be effectively used for clustering and validating not only DNA microarray expression analysis applications but also other biomedical and physical data with no limitations. AVAILABILITY: The program is freely available for non-profit use on request at http://www.cs.tcd.ie/Nadia.Bolshakova/Machaon.html CONTACT: Nadia.Bolshakova@cs.tcd.ie.