Assessing the Value of the Zebrafish Conditioned Place Preference Model for Predicting Human Abuse Potential.

Regulatory agencies recommend that centrally active drugs are tested for abuse potential before approval. Standard preclinical assessments are conducted in rats or non-human primates (NHPs). This study evaluated the ability of the zebrafish conditioned place preference (CPP) model to predict human abuse outcomes. Twenty-seven compounds from a variety of pharmacological classes were tested in zebrafish CPP, categorized as positive or negative, and analyzed using standard diagnostic tests of binary classification to determine the likelihood that zebrafish correctly predict robust positive signals in human subjective effects studies (+HSE) and/or Drug Enforcement Administration drug scheduling. Results were then compared with those generated for rat self-administration and CPP, as well as NHP self-administration, using this same set of compounds. The findings reveal that zebrafish concordance and sensitivity values were not significantly different from chance for both +HSE and scheduling. Although significant improvements in specificity and negative predictive values were observed for zebrafish relative to +HSE, specificity without sensitivity provides limited predictive value. Moreover, assessments in zebrafish provided no added value for predicting scheduling. By contrast, rat and NHP models generally possessed significantly improved concordance, sensitivity, and positive predictive values for both clinical measures. Although there may be predictive value with compounds from specific pharmacological classes (e.g., µ-opioid receptor agonists, psychostimulants) for zebrafish CPP, altogether these data highlight that using the current methodology, the zebrafish CPP model does not add value to the preclinical assessment of abuse potential.

An intronic polymorphism in the porcine IRF7 gene is associated with better health and immunity of the host during Sarcocystis infection, and affects interferon signalling.

Interferon regulatory factor 7 (IRF7), as a key regulator of type I interferon response, plays an important role during innate response against viral infection. Although well conserved across species, the structure of IRF7 and its function during parasite infection are not well documented in farm animals, such as the pig. To bridge this gap, we have determined the porcine IRF7 gene structure and identified two intronic single nucleotide polymorphisms (SNPs), SNP g.748G>C and SNP g.761A>G, in commercial pig breeds. The distribution of SNP g.761A>G in multiple breeds suggested that it was in Hardy-Weinberg equilibrium and allowed us to map it at the top of SSC2. We found that during Sarcocystis miescheriana infection, the G allele was associated with high lymphocyte levels (P < 0.02), reduced drop in platelet levels (P < 0.002) and IgG1-Th2-dominated response (P < 0.05). This suggests that the G allele was associated with better health and immunity of the host during Sarcocystis infection. Furthermore, we have also provided suggestive evidence that the G allele of SNPc.761A>G enhances the transactivation activity of IRF7, possibly by improving IRF7 transcript splicing of intron-3. These findings would suggest that IRF7, as a transcriptional regulator, is involved in the defence mechanism against a larger spectrum of pathogens, and in more host species, than initially anticipated.

Dinuclear triple stranded phenyl-spaced 1,3-bis-ß-diketonato lanthanide(iii) complexes: synthesis, structures and spectroscopy.

The synthesis and crystal structures of a series of Ln(iii) (Ln = Gd, Nd, Yb as well as Y) complexes of a bis-ß-diketone ligand incorporating a 1,3-substituted phenyl ring between its two ß-diketone domains (1,1'-(1,3-phenylene)bis(4,4-dimethylpentane-1,3-dione), H2L1) is reported. The crystal structures show the complexes are seven coordinate in the solid state with the general formula [Ln2L13(solvent)2]. The photophysical properties of the complexes were explored in both solution and the solid state, which show an increase in coordination number in solution.

Comparison of endotoxins and cutaneous burn toxin as immunosuppressants.

Endotoxins of E. coli, S. typhosa and Ps. aeruginosa were injected i.p. into mice a few days before administration of the antigen sheep erythrocytes (SE). Antibody-forming cells (AFC) to SE were later enumerated in relation to dose of endotoxin given. In comparison a toxic lipid protein isolated from burned skin (cutaneous burn toxin or CBT) was similarly applied and found to be more inhibitory of the immune response than any of the three endotoxins. Considering the 50 per cent inhibitory doses on a molar basis CBT was found to be 1000 fold more immunosuppressive than the most inhibitory endotoxin. As the immune suppression which follows severe thermal injury involves failure of interleukin 2 (IL2) function, as a critical index of survival, the CBT was tested for its effects on the culture of a human IL2-dependent cell line in the presence of IL2. CBT inhibited the growth of these cells, however, endotoxin had no effect on their proliferation. Thus CBT, which arises by a thermally induced polymerization of skin lipid protein, is specific to burn injury and has a direct inhibitory effect on the immune response.

Temperature-dependent characteristics of Teflon membranes used in mass spectrometry.

In vitro information is provided on the gas partial pressure/temperature relationship and on the membrane permeability/temperature relationship, when a special type of Teflon membrane is used with a mass spectrometer. Monitored pressures of oxygen, nitrogen, and carbon dioxide in an equilibrating gas mixture decreased with stepwise decreases in temperature from 37 to 18 degrees C. Permeability of membranes to all gases also decreased with decreasing temperature. Observed partial pressure changes were apparently due to temperature-induced changes in Teflon membrane permeability characteristics.

Chemiluminescence in human whole blood: modulation by the cocarcinogens phorbol diester and polychlorobiphenyls.

A human whole blood chemiluminescence (CL) assay was established using zymosan as cell activator. Aroclor 1254 was found to inhibit this CL response in a direct linear relation to its concentration, (50% inhibitory dose, (ID50) equal to 5 x 10(-4)M) in diluted blood samples of 10 normal human subjects. In comparison the ID50 of other inhibitors was 1.3 x 10(-3)M for ethylenediamine tetraacetic acid, 3.3 x 10(-3)M for ascorbic acid, 4 x 10(-3)M for reduced glutathione, 1.2 x 10(-1)M for ethanol, 2.5 x 10(-1)M for methanol and 3.7 x 10(-1)M for dimethyl sulfoxide. Using 12-o-tetradecanoyl-phorbol-13-acetate (TPA) as cell activator the CL response was likewise inhibited by Aroclor 1254 with an ID50 of 4.5 x 10(-4)M. However, it was found that Aroclor 1254 alone has a stimulatory CL effect on otherwise unactivated cells. To compare the mechanisms involved in the CL elicited by the three stimulants zymosan, TPA and Aroclor 1254, the CL signal was measured in the presence of cytochalasin B. Cytochalasin B inhibited zymosan-induced CL, had a smaller inhibitory effect on TPA-induced CL but it could augment the CL response initiated by Aroclor 1254. This pattern of responses implicates Aroclor 1254 in the activation of eicosanoid metabolism as it matches the differential responses reported for arachidonic acid.