Spectroscopic studies of copper, silver and gold-metallothioneins

Metallothionein is a ubiquitous protein with a wide range of proposed physiological roles, including the transport, storage and detoxification of essential and nonessential trace metals. The amino acid sequence of isoform 2a of rabbit liver metallothionein, the isoform used in our spectroscopic studies, includes 20 cysteinyl groups out of 62 amino acids. Metallothioneins in general represent an impressive chelating agent for a wide range of metals. Structural studies carried out by a number of research groups (using (1)H and (113)Cd NMR, X-ray crystallography, more recently EXAFS, as well as optical spectroscopy) have established that there are three structural motifs for metal binding to mammalian metallothioneins. These three structures are defined by metal to protein stoichiometric ratios, which we believe specifically determine the coordination geometry adopted by the metal in the metal binding site at that metal to protein molar ratio. Tetrahedral geometry is associated with the thiolate coordination of the metals in the M(7)-MT species, for M = Zn(II), Cd(II), and possibly also Hg(II), trigonal coordination is proposed in the M(11-12)-MT species, for M = Ag(I), Cu(I), and possibly also Hg(II), and digonal coordination is proposed for the metal in the M(17-18)-MT species for M = Hg(II), and Ag(I). The M(7)-MT species has been completely characterized for M = Cd(II) and Zn(II). (113)Cd NMR spectroscopic and x-ray crystallographic data show that mammalian Cd(7)-MT and Zn(7)-MT have a two domain structure, with metal-thiolate clusters of the form M(4)(S(cys))(11) (the alpha domain) and M(3)(S(cys))(9) (the beta domain). A similar two domain structure involving Cu(6)(S(cys))(11) (alpha) and Cu(6)(S(cys))(9) (beta) copper-thiolate clusters has been proposed for the Cu(12)-MT species. Copper-, silver- and gold-containing metallothioneins luminesce in the 500-600 nm region from excited triplet, metal-based states that are populated by absorption into the 260-300 nm region of the metal-thiolate charge transfer states. The luminescence spectrum provides a very sensitive probe of the metal-thiolate cluster structures that form when Ag(I), Au(I), and Cu(I) are added to metallothionein. CD spectroscopy has been used in our laboratory to probe the formation of species that exhibit well-defined three-dimensional structures. Saturation of the optical signals during titrations of MT with Cu(I) or Ag(I) clearly show formation of unique metal-thiolate structures at specific metal:protein ratios. However, we have proposed that these M=7, 12 and 18 structures form within a continuum of stoichiometries. Compounds prepared at these specific molar ratios have been examined by X-ray Absorption Spectroscopy (XAS) and bond lengths have been determined for the metal-thiolate clusters through the EXAFS technique. The stoichiometric ratio data from the optical experiments and the bond lengths from the XAS experiments are used to propose structures for the metal-thiolate binding site with reference to known inorganic metal-thiolate compounds.     

An anomaly-based approach to the analysis of the social behavior of VoIP users

In this paper we present the results of a study we recently conducted by analyzing a large data set of VoIP Call Detail Records (CDRs), provided by an Italian telecom operator. The objectives of this study were twofold: (i) first, to provide a representation of users behavior, as well as of their mutual interaction and communication patterns, allowing to identify certain easily separable user categories; and (ii) second, to design and implement a framework calculating such a representation starting from CDR, capable of operating within certain time constraints, and grouping users using unsupervised techniques.The paper shows how we can reliably identify behavioral patterns associated with the most common anomalous behaviors of VoIP users. It also exploits the expressive power of relational graphs in order to both validate the results of the unsupervised analysis and ease their interpretation by human operators.     

Climate change benefits and energy supply benefits as determinants of acceptance of nuclear power stations: Investigating an explanatory model

Several countries are currently discussing whether they will rebuild their nuclear power stations in order to continue this type of energy production in the future. The public, with its own opinion about nuclear power stations, has an influential voice in this discussion. As a result, policy makers and nuclear scientists are interested in the public's perception of nuclear power and in what determines this perception. We therefore examined an explanatory model of the public's acceptance of nuclear power based on a telephone survey among a representative sample in Switzerland. The model included such factors as risk perception, benefit perception, affective feelings, and social trust. Moreover, we distinguished between two types of benefit perception: benefit for the climate and a secure energy supply. The model fitted very well to our data and explained acceptance very well. Acceptance was mainly influenced by perceived benefits for a secure energy supply and, to a lesser extent, both by perceived benefits for the climate and by risk perception. Affective feelings about nuclear power appeared to be a central factor in the model. Implications for communication about nuclear power stations and for further research are discussed.     

An investigation of scotopic threshold-versus-illuminance curves for the analysis of color-matching data

Stiles developed a method for estimating whether rod-photoreceptor activity contributed to color-matching data. This method requires knowledge of the rod threshold at the illuminance levels of lights being viewed. Stiles suggested that the Aguilar and Stiles scotopic threshold-versus-illuminance (TVI) curve could serve as a convenient approximation of the rod threshold. Stiles' method was intended for use in color-matching experiments where the stimuli are presented side by side, but, in the Aguilar and Stiles experiment, detection thresholds were measured for temporally pulsed lights. This study compares rod-sensitivity with detection and side-by-side adjustment tasks. The illumination level of the test and background light was kept below the absolute threshold of the short-wavelength sensitive cones, because there is evidence that cone signals may interact with the scotopic TVI function. The threshold data fell mainly within the range of the Aguilar and Stiles observers. There were two notable deviations from the Aguilar and Stiles curve: (1) consistent with previous reports, the scotopic TVI function had a slope shallower than the Aguilar and Stiles curve when measured with a short-wavelength background; (2) thresholds measured from a dark surround with a spatial adjustment task were higher than the Aguilar and Stiles thresholds even though the slopes were the same. © 1996 John Wiley & Sons, Inc.     

Intracranial self-stimulation of the dorsal raphe sensitizes psychostimulant locomotion.

The authors hypothesized that repeated rewarding electrical stimulation of the dorsal raphe can produce behavioral sensitization to psychostimulants. Groups of male rats were implanted with a stimulation electrode and preexposed to brain stimulation at parameters set to equate rewarding effectiveness across rats. Control groups were implanted with an electrode but never stimulated, or not implanted at all. Twenty-four hours after the 12th self-stimulation session, all groups were challenged with amphetamine (0.5 mg/kg, ip), nicotine (0.2 mg/kg, sc), or saline, and locomotor activity was measured for 1 hr. Locomotor responses to amphetamine and to nicotine were significantly greater in rats preexposed to brain stimulation. These findings suggest at least partial overlap of underlying substrates. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Mucosal addressin cell adhesion molecule-1 (MAdCAM-1). Its binding motif for alpha 4 beta 7 and role in experimental colitis

The integrin alpha 4 beta 7 and mucosal addressin cell adhesion molecule-1 (MAdCAM-1) are molecules involved in the normal recirculation of lymphocytes between the blood and the gastrointestinal tract. These molecules may play a complementary and significant role in animal models of colitis. We have investigated the structural interaction between alpha 4 beta 7 and MAdCAM-1. Site-directed mutagenesis studies of the MAdCAM-1 molecule has led to the identification of the amino acid residue (LDT) in the loop between beta strands C and D of the Ig-superfamily-like folds being involved in the adhesive and cell activation functions of MAdCAM-1 with alpha 4 beta 7.     

Perception de contr?le, désir de contr?le et santé psychologique.

Studied interactions among perceptions of control, desire for control, and mental health. Human Ss: 224 normal male and female Canadian adults (university students) (mean age 24 yrs). Ss completed questionnaires assessing perceptions of control, desire for control, psychological distress, feelings of helplessness, and symptoms of depression and anxiety. Interactions among these variables were analyzed. Tests used: Several instruments, including the Spheres of Control Scale (D. L. Paulhus, 1983). (English abstract) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Internalization of basic fibroblast growth factor (bFGF) in cultured endothelial cells: role of the low affinity heparin-like bFGF receptors

We have shown (Presta et al., Cell Regul., 2:719-726, 1991) that a long-lasting interaction of basic fibroblast growth factor (bFGF) with endothelial GM 7373 cells is required to induce cell proliferation. In the present work, we have investigated the interaction of 125I-bFGF with GM 7373 cells, its pathway of internalization, and its intracellular fate under the same experimental conditions previously utilized to assess the mitogenic activity of the growth factor. Cell cultures were incubated with 10 ng/ml 125I-bFGF for 2 h at 4 degrees C. Then, cells were shifted to 37 degrees C without changing the medium. A rapid down-regulation of high affinity sites, paralleled by a rapid internalization of 125I-bFGF, was observed during the first 1-2 h at 37 degrees C. After 6-8 h, also low affinity sites down-regulate. This was paralleled by a continuous internalization of 125I-bFGF and by a slow disappearance of the growth factor from the culture medium. This suggests that GM 7373 cells activate, when exposed to bFGF for a long period of time, a late internalization pathway mediated by low affinity sites. This hypothesis was supported by the following experimental evidence: 1) soluble heparin inhibited the prolonged internalization of 125I-bFGF and its binding to low affinity sites with the same potency; 2) treatment of GM 7373 cells with heparinase, which removes most of the low affinity sites, also inhibited the prolonged internalization of 125I-bFGF. 125I-bFGF internalized via low affinity sites was partially protected from lysosomal degradation. This was the case also when 125I-bFGF was internalized in the presence of soluble heparin, suggesting that the complexes bFGF-cell surface glycosaminoglycan and bFGF-soluble heparin are maintained during the internalization of the growth factor. Moreover, the capacity of soluble heparin to inhibit the mitogenic activity of bFGF also when added to cell cultures several hours after the growth factor indicates that the requirement for a prolonged interaction of bFGF with GM 7373 cells in order to induce cell proliferation might be related to the late internalization of the growth factor via low affinity sites.