Acamprosate and alcohol: I. Effects on alcohol intake following alcohol deprivation in the rat

Acamprosate (calcium-acetyl homotaurinate) is a new compound in the treatment of alcoholism. Its efficacy has been proven in several clinical trials and registration is now pending in most European countries. The basic mechanisms by which acamprosate elicits its anti-craving action, thereby leading to reduced relapse rates, is not known at the moment. In the present study we describe a rat model of long-term alcohol-drinking which mimics relapse behavior in human alcoholics. The effect of acamprosate was studied in this model. Wistar rats had a free choice between water and alcohol solutions of different concentrations (5, 10, 20% v/v). After two months of continuous alcohol access, rats were deprived of alcohol for three days. Following this deprivation phase, all alcohol solutions were presented again. This procedure was repeated monthly for the following six months. The rats consumed 3.5 +/- 0.3 g/kg alcohol a day. After alcohol deprivation, alcohol intake rose to 5.2 +/- 0.3 g/kg per day resulting in blood alcohol levels of 30 +/- 6 mg/dl. Interestingly, the addition of quinine to the alcohol solutions or the additional presentation of a 5% sucrose solution did not affect the alcohol-deprivation effect after eight months of this intermittent alcohol exposure. However, when acamprosate (50-200 mg/kg i.p.) was administered twice daily, alcohol-drinking following an alcohol-deprivation phase was decreased dose dependently. Given at the highest dose alcohol intake even dropped significantly below baseline drinking. Together, these results show that acamprosate effectively diminishes the alcohol-deprivation effect. Furthermore, the described model seems to be a suitable animal model to screen compounds for their anti-relapse properties and subsequently for their anti-craving action.     

Vision assistance in scenes with extreme contrast

The performance and architecture of a high dynamic range camera (HDRC) chip and the conceptional advantages for its adaptation to image processing systems in traffic environments are discussed. The HDRC chip was developed with 64×64 pixels using a standard digital 1.2-μm CMOS technology. It is shown that the implementation proves the functionality and indicates the system performance of a highly dynamic range camera is feasible     

Analysis of heregulin symmetry by weighted evolutionary tracing

Heregulins are members of the protein family of EGF-like growthand differentiation factors. The primary cell-surface targetsof heregulins are heterodimers of the EGF-receptor homolog HER2with either HER3 or HER4. We used a weighted evolutionary traceanalysis to identify structural features that distinguish theEGF-like domain (hrg) of heregulins from other members of theEGF family. In this analysis, each amino acid sequence is weightedaccording to its uniqueness and the variability in each positionis assigned by an amino acid substitution matrix. Conservedresidues in heregulin that are variable in other EGF-like domainsare considered possible specificity-conferring residues. Thisanalysis identifies two clusters of residues at the foot ofthe boot-shaped hrg domain. The residues in one cluster arerecruited from the N-terminus; those in the other are from the-loop region and show a weak sequence similarity to the N-terminalresidues at the opposite side of the boot. The remaining residueswith high conservation scores distribute themselves into thesetwo distinct surfaces on hrg. This pseudo-twofold symmetry andthe presence of two distinct interfaces may reflect the preferenceof hrg for heterodimeric versus homodimeric HER complexes.     

Interfacial Tension; a Stabilizing Factor for Janus Emulsions of Silicone Bixa Orellana Oils

The destabilization process was investigated for a Janus emulsion of silicone and Bixa Orellana oils stabilized by polyoxyethylene sorbitan monooleate (Tw 80) and carboxymethyl cellulose. The emulsion stabilized with Tw 80 showed significant and fast creaming, a process that was prevented by the addition of the polymer. During the extensive coalescence of the emulsions stabilized by Tw 80, the Janus topology was retained for months of storage until, finally, separation of the oils occurred. This result strongly indicates an unexpected stabilizing action of the i nterfacial free energy. This conclusion was supported by a calculation for a realistic model system of the interfacial energy difference between two cases of coalescence. In the first case, the two coalescing Janus drops united into a larger Janus drop, while in the second case two drops formed, each with only one oil. The first case gave a spontaneous reaction (reduced interfacial energy), while the second one meant an increase of energy, i.e. it cannot happen without adding energy. The authors are aware that this stabilization is a new phenomenon in emulsion science with potential ramifications in future emulsion technology. However, it is essential to realize that the stabilization is of temporary occurrence in the destabilization process, and the free energy to give a final emulsion state with separated oils is overwhelmingly dominant. In short, Janus emulsions will, in the end, separate into layers of the liquids, like all emulsions.     

Evaluation of potential interfering agents on in vitro methods for the determination of the antioxidant capacity in anthocyanin extracts

The evaluation of the effects of sugars, metals, acids and other antioxidants on the in vitro antioxidant capacity of purified anthocyanin extract by different techniques was the purpose of this study. Three methods and the ways of expressing their results were evaluated: ABTS_TEAC (capacity equivalent to Trolox, by ABTS (2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulphonic acid))), DPPH_TEAC (capacity equivalent to Trolox, by DPPH (2,2‐diphenyl‐1‐picrylhydrazyl)), DPPH_EC50 (50% reduction in the radical, by DPPH), DPPH_%Sca (reduction in the scavenging capacity, by DPPH), FRAP_TEAC (capacity equivalent to Trolox, by FRAP (reduction power of iron)) and FRAP_EC50 (50% reduction in the radical, by FRAP). The way of expressing DPPH and FRAP results as EC50 showed the greater interfering extent, mainly when the medium contained tartaric and ascorbic acids. The most coherent method was ABTS_TEAC in which only ascorbic acid interfered. Ascorbic acid was shown to interfere in all methods; thus, it must be removed prior to determining the in vitro antioxidant capacity of anthocyanins in food materials.     

Incidence of Listeria spp. and Salmonella spp. in horsemeat for human consumption

The incidence of Salmonella spp., Listeria spp. and Listeria monocytogenes in horsemeat for human consumption was investigated. One-hundred and twenty-one samples of frozen horsemeat collected from two Brazilian abattoirs were analysed over a period of 1 year. Twenty-two samples (18.2%) were positive for Listeria spp. with nine (7.4%) containing L. monocytogenes. None of the samples harbored Salmonella spp.     

Behavioral, physiological, and neuroendocrine stress responses and differential sensitivity to diazepam in two Wistar rat lines selectively bred for high- and low-anxiety-related behavior

Two Wistar rat lines, selectively bred for high-anxiety-related behavior (HAB) and low-anxiety-related behavior (LAB) in the elevated plus-maze test, were tested for the susceptibility of their behavioral characteristics to anxiolytic treatment and for their endocrine and physiological reactivity to different stressors. Injection of 1 mg/kg diazepam failed to affect line differences in coping strategy but resulted in a marked (20-fold) decrease in plus-maze anxiety in HAB rats; whereas, the anxiolytic effect was less pronounced in LAB animals. Biotelemetrical measurements revealed that HAB and LAB rats do not significantly differ in their baseline body temperature, locomotor activity, food and water intake, or in stress-induced alterations of the diurnal rhythms in these parameters. However, line differences were found in acute changes in body temperature and locomotor activity following stress exposure, LAB rats responding with a greater, albeit shorter, increase in body temperature and activity than HAB animals. Basal ACTH and corticosterone plasma levels as well as pituitary reactivity to intravenously administered CRH (40 ng/kg) were similar in both lines, although, especially in response to plus-maze exposure, HAB rats tended toward higher ACTH secretion than LAB rats. These data confirm that animals with high or low basal levels of anxiety may be a promising model for studying the mechanisms of action of anxiolytic substances. Nevertheless, the endocrine findings support the notion that the reactivity of the hypothalamo-pituitary-adrenocortical system and anxiety-related behavior can be regulated independently.     

Multiple sulfa compounds: high-pressure liquid chromatographic assay and mobile phase correlation

A mixture of sulfacetamide, sulfathiazole, and sulfabenzamide was used to develop a rapid high-pressure liquid chromatographic assay. In addition, this study provided a means to develop concepts relating solvent molar polarization parameters and retention times. A linear correlation between molar polarization and retention time was observed and will permit reasonably rapid predictions about the dependent variable.     

Cytotoxicity and specificity of directed toxins composed of diphtheria toxin and the EGF-like domain of heregulin beta1

As a step in the design of directed toxins, aimed at cells that overexpress HER receptors, particularly breast carcinoma cells, we studied the properties of a chimera of diphtheria toxin (DT) and heregulin beta1. The EGF-like growth hormone heregulin is a ligand for the HER3 and HER4 receptors and their heterodimers with HER2. The 60-residue EGF-like domain (hrg) of heregulin elicits a biological response and binds to these receptors primarily through its N terminus. We tested a fusion protein in which hrg replaces the C-terminal receptor-binding domain of DT (DT(389)hrg) and an alternative design in which this domain is fused to the N terminus of DT(389). Of those two constructs, the N-terminal fusion was not active as a directed toxin but elicited a growth response. The C-terminal fusion of hrg to DT(389) yielded a functional toxin and showed cell line specific cytotoxicity that is consistent with heregulin specificity. The binding of hrg to its cognate receptor is not impaired as shown by receptor activation, direct binding, and competition with free hrg. Cytotoxicity is dependent on high-affinity binding of DT(389)hrg to HER3 and HER4 receptors and is not mediated by HER2 overexpression alone. For those cell lines exhibiting high-affinity binding sites, the level of cytotoxicity correlates with the rate of internalization. Thus DT(389)hrg chimeras offer a possible avenue toward directed toxins against cells that overexpress HER receptors.