Exploring the unique pharmacology of a novel opioid receptor, ZFOR1, using molecular modeling and the `message-address' concept

Previous studies have probed the structural basis of ligandselectivity in the mu, delta and kappa opioid receptors throughthe application of molecular modeling techniques in concertwith the `message–address' concept. Here, this approachwas used in an attempt to rationalize the unique pharmacologicalprofile of a recently cloned novel opioid receptor, ZFOR1 (ZebraFishOpioid Receptor 1). Specifically, a model of the transmembranedomains of ZFOR1 was constructed and used to explore the bindingmodes of various prototypical opioid ligands. The results showthat the `message' portion of the binding pocket of ZFOR1 ishighly conserved; hence, the binding modes of non-selectiveopioid ligands are well preserved. In contrast, a small numberof variant residues at the extracellular end of the bindingpocket, particularly Lys288 (VI:26) and Trp304 (VII:03), areshown to create adverse steric interactions with all delta andkappa selective ligands examined, thereby disrupting their bindingmodes. These results are consistent with, and serve as an explanationfor, the observed pharmacology of this receptor, lending supportto both the validity of the `message–address' conceptitself and to the use of molecular modeling approaches in itsapplication.     

Evaluation of the combined solar TiO2/photo-Fenton process using multivariate analysis.

The effect of combining the photocatalytic processes using TiO2 and the photo-Fenton reaction with Fe3+ or ferrioxalate as a source of Fe2+ was investigated in the degradation of 4-chlorophenol (4CP) and dichloroacetic acid (DCA) using solar irradiation. Multivariate analysis was used to evaluate the role of three variables: iron, H2O2 and TiO2 concentrations. The results show that TiO2 plays a minor role when compared to iron and H2O2 in the solar degradation of 4CP and DCA in the studied conditions. However, its presence can improve TOC removal when H2O2 is totally consumed. Iron and peroxide play major roles, especially when Fe(NO3)3 is used in the degradation of 4CP. No significant synergistic effect was observed by the addition of TiO2 in this process. On the other hand, synergistic effects were observed between FeOx and TiO2 and between H2O2 and TiO2 in the degradation of DCA.     

Clinical outcome of mild fetal ventriculomegaly

The presence of antibodies against neural antigens was investigated in the serum of patients with schizophrenia, major depression and normal controls. Different immunological abnormalities, humoral and cellular, were reported in schizophrenia and major depression. The pathogenesis of schizophrenia is multifactorial. An autoimmune mechanism was suggested as a possible factor. We tested the serum of 26 patients with schizophrenia, eight patients with major depression and 22 normal controls. The serum samples were tested for antibody binding to protein extracts of IMR-32 neuroblastma cell line using Western blot analysis. Immunoglobulins of eight patients with schizophrenia (30.71%) reacted with a protein of 80-85 kDa. Serum samples from subjects of other groups did not react with this protein. Sera of all patients with major depression but one, and all normal controls reacted with HSP 60 kDa to different extent. This is an apparent discrepancy with the findings of Kilidireas et al. [Kilidireas, K., Latov, N., Strauss, D.H., Gorig, A.D., Hashim, G.A., Gorman, J.M., Sadig, S.A., 1992. Antibodies to the human 60 kDa heat shock protein in patients with schizophrenia. Lancet 340, 569-572.] who demonstrated the presence of antibodies against HSP 60 kDa in 44% of patients with schizophrenia tested and 8% of normal subjects. HSP 60 kDa is an antigen of many pathogens and antibodies against it might be a result of an infection and cannot be a good indicator for an autoimmune process. The presence of antibodies against a protein of 80-85 kDa should be investigated as a possible specific indicator.     

Evaluation of the influences of solution path length and additives concentrations on the solar photo-Fenton degradation of 4-chlorophenol using multivariate analysis

This study reports the photodegradation of 4-chlorophenol (4-CP) in aqueous solution by the photo-Fenton process using solar irradiation. The influence of solution path length, and Fe(NO(3))(3) and H(2)O(2) concentrations on the degradation of 4-CP is evaluated by response surface methodology. The degradation process was monitored by the removal of total organic carbon (TOC) and the release of chloride ion. The results showed a very important role of iron concentration either for TOC removal or dechlorination. On the other hand, a negative effect of increasing solution path length on mineralization was observed, which can be compensated by increasing the iron concentration. This permits an adjustment of the iron concentration according to the irradiation exposure area and path length (depth of a tank reactor). Under optimum conditions of 1.5 mM Fe(NO(3))(3), 20.0 mM H(2)O(2) and 4.5 cm solution path length, 17 min irradiation under solar light were sufficient to reduce a 72 mg CL(-1) solution of 4-CP by 91%.     

Evaluation of cell proliferation through a polymerase chain reaction-based cyclin A test

Lipid binding properties of cytochrome c (cyt c) were investigated by using semisynthetic mutant protein having amino acid substitution on the evolutionary invariant residue Arg91. We demonstrate here that the membrane binding properties of cyt c are dramatically altered by substituting norleucine for the invariant Arg91. More specifically, while the binding of this mutant to liposomes per se is indistinguishable from the wild type protein, it completely lacks the ability of the native cyt c to rupture liposome bilayers.     

Combined elastoplastic and limit analysis via restricted basis linear programming

In many engineering situations rigid-perfectly plastic limit analysis turns out to be insufficient and unreliable. On the other hand, elastic-plastic “historical” analysis is often laborious. For trusslike frames and suitably discretized structures the former analysis is traditionally cast into the mathematical model of linear programming; the latter has led to various formulations and solution methods explicitly or implicitly related to quadratic programming.In this paper the complete stress and strain response of elastic-perfectly plastic structures subjected to proportional loading up to collapse is determined by a method essentially consisting of a linear programming procedure supplemented by an additional rule which enforces a complementarity relation among variables at each pivotal step (“restricted basis entry”). Possible local unloadings are allowed for by means of suitable modifications of the current tableau. The method proposed provides a whole “exact” deformation history of the structure, the safety factor and the collapse mechanism. Lack of uniqueness in the deformation history and “limited mechanisms” prior to collapse are discussed. Nonholonomic (irreversible) analysis and holonomic (reversible nonlinear-elastic) analysis are compared.The computational effort is assessed and checked by examples and turns out to be roughly equivalent to that required by the familiar rigid-plastic limit analysis by the static approach via standard linear programming. Extensions of the method to strain-hardening behaviour, to nonproportional loading processes and to broader categories of structures are envisaged.     

Opposing regulation of tau protein levels by ionotropic and metabotropic glutamate receptors in human NT2 neurons

Human NT2-N neurons derived from retinoic acid treatment of the NTera 2 cell line were used to determine the consequences of ionotropic glutamate receptor (iGluR) hyperstimulation and possible modulatory role(s) exerted by metabotropic glutamate receptor (mGluR) activation. We found that NT2-N neurons express the NR1 subunit of N-methyl-D-aspartate (NMDA) iGluRs and mRNA encoding the 1a isoform of mGluRs. A 15 min pulse with 100 microM NMDA induced an increase in the levels of tau proteins in NT2-N cells. This effect was prevented by incubating NT2-N neurons in the presence of the mGluR agonist (1 S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD). This phenomenon was related, in terms of doses and time, with the observed 1S,3R-ACPD-mediated protection against NMDA-induced NT2-N cell death. Our findings suggest that iGluRs and mGluRs might participate in the control of human neuron viability by differentially affecting the expression of tau proteins.     

Optimization heuristics for determining internal rating grading scales

Basel II imposes regulatory capital on banks related to the default risk of their credit portfolio. Banks using an internal rating approach compute the regulatory capital from pooled probabilities of default. These pooled probabilities can be calculated by clustering credit borrowers into different buckets and computing the mean PD for each bucket. The clustering problem can become very complex when Basel II regulations and real-world constraints are taken into account. Search heuristics have already proven remarkable performance in tackling this problem. A Threshold Accepting algorithm is proposed, which exploits the inherent discrete nature of the clustering problem. This algorithm is found to outperform alternative methodologies already proposed in the literature, such as standard k-means and Differential Evolution. Besides considering several clustering objectives for a given number of buckets, we extend the analysis further by introducing new methods to determine the optimal number of buckets in which to cluster banks’ clients.     

Verocytotoxin-producing Escherichia coli O157 in minced beef and dairy products in Italy

A total of 3879 samples of foodstuffs were examined for the presence of Verocytotoxin-producing Escherichia coli O157 (VTEC O157). The survey was conducted by 9 of the 10 Italian Veterinary Public Health Laboratories. Samples were collected between May 2000 and September 2001 in 14 regions and comprised 931 minced beef specimens and 2948 dairy products (DP) with less than 60 days of ripening. The DP included 657 pasteurised and 811 unpasteurised bovine DP, 477 pasteurised and 502 unpasteurised ovine DP, and 501 water-buffalo's milk mozzarella cheese. Samples were collected at retail level, from plants processing minced beef and dairy plants and from farms directly manufacturing cheeses. All the samples were tested using a sensitive procedure based on ISO/DIS 16654:1999 (later ISO 16654:2001), which includes an immunomagnetic separation step. A preliminary inter-laboratory trial was organised with artificially contaminated samples to assess the ability of all the participating laboratories to isolate E. coli O157 by the established procedure. VTEC O157 was isolated from four (0.43%) of the minced beef samples, collected in four different regions and during different months, but was not detected in any of the dairy products. E. coli O157 VT-eae+ was isolated from one raw cow's milk cheese. This survey provided national data on the presence of VTEC O157 in foodstuffs, demonstrating a low prevalence of the organism. The survey also encouraged updating of knowledge and procedures on VTEC O157 in laboratories with official responsibility for microbiological testing of foods of animal origin.     

Hypothalamic NPY-Y1R Interacts with Gonadal Hormones in Protecting Female Mice against Obesity and Neuroinflammation

We previously demonstrated that Npy1r^rfb mice, which carry the conditional inactivation of the Npy1r gene in forebrain principal neurons, display a sexually dimorphic phenotype, with male mice showing metabolic, hormonal and behavioral effects and females being only marginally affected. Moreover, exposure of Npy1r^rfb male mice to a high-fat diet (HFD) increased body weight growth, adipose tissue, blood glucose levels and caloric intake compared to Npy1r^2lox male controls. We used conditional knockout Npy1r^rfb and Npy1r^2lox control mice to examine whether forebrain disruption of the Npy1r gene affects susceptibility to obesity and associated disorders of cycling and ovariectomized (ovx) female mice in a standard diet (SD) regimen or exposed to an HFD for 3 months. The conditional deletion of the Npy1r gene increased body weight and subcutaneous white adipose tissue weight in both SD- and HFD-fed ovx females but not in cycling females. Moreover, compared with ovx control females on the same diet regimen, Npy1r^rfb females displayed increased microglia number and activation, increased expression of Neuropeptide Y (NPY)-immunoreactivity (IR) and decreased expression of proopiomelanocortin-IR in the hypothalamic arcuate nucleus (ARC). These results suggest that in the ARC NPY-Y1R reduces the susceptibility to obesity of female mice with low levels of gonadal hormones and that this effect may be mediated via NPY-Y1R ability to protect the brain against neuroinflammation.