Current status, between-year comparisons and maternal transfer of organohalogenated compounds (OHCs) in Arctic char (Salvelinus alpinus) from Bjørnøya, Svalbard (Norway).

High levels of organohalogenated compounds (OHCs) have been found in Arctic char from Lake Ellasjøen at Bjørnøya (Svalbard, Norway) compared to char from other arctic lakes. The first aim of the study was to investigate the OHC status, contaminant profile, and partitioning of OHCs between muscle and ovary tissue in spawning female char from the high-polluted Lake Ellasjøen and the low-polluted Lake Laksvatn. The second aim was to investigate if OHC levels in muscle tissue have changed over time. Between-lake comparisons show that the muscle levels (lipid weight) of hexachlorobenzene (HCB), chlordanes (∑CHLs), mirex, dichlorodiphenyltrichloroethanes (∑DDTs) and polychlorinated biphenyls (∑PCBs) were up to 36 times higher in char from Ellasjøen than in Laksvatn, and confirm that the char from Ellasjøen are still heavily exposed compared to char from neighboring lake. A higher proportion of persistent OHCs were found in Ellasjøen compared to Laksvatn, while the proportion of the less persistent OHCs was highest in Laksvatn. A between-year comparison of OHC levels (i.e., HCB, DDTs, PCBs) in female and male char shows higher levels of HCB in female char from Ellasjøen in 2009/2012 compared to in 1999/2001. No other between-year differences in OHC levels were found. Due to small study groups, findings associated with between-year differences in OHC levels should be interpreted with caution. OHCs accumulate in the lipid rich ovaries of spawning females, resulting in up to six times higher levels of OHCs in ovaries compared to in muscle (wet weight). The toxic equivalent (TEQ)-value for the dioxin-like PCBs (PCB-105 and -118) in ovaries of the Ellasjøen char exceeded levels associated with increased egg mortality in rainbow trout (Oncorhynchus mykiss). Hence, we suggest that future studies should focus on the reproductive health and performance abilities of the high-exposed population of char inhabiting Lake Ellasjøen.

Towards an alternative testing strategy for nanomaterials used in nanomedicine: lessons from NanoTEST.

In spite of recent advances in describing the health outcomes of exposure to nanoparticles (NPs), it still remains unclear how exactly NPs interact with their cellular targets. Size, surface, mass, geometry, and composition may all play a beneficial role as well as causing toxicity. Concerns of scientists, politicians and the public about potential health hazards associated with NPs need to be answered. With the variety of exposure routes available, there is potential for NPs to reach every organ in the body but we know little about the impact this might have. The main objective of the FP7 NanoTEST project ( www.nanotest-fp7.eu ) was a better understanding of mechanisms of interactions of NPs employed in nanomedicine with cells, tissues and organs and to address critical issues relating to toxicity testing especially with respect to alternatives to tests on animals. Here we describe an approach towards alternative testing strategies for hazard and risk assessment of nanomaterials, highlighting the adaptation of standard methods demanded by the special physicochemical features of nanomaterials and bioavailability studies. The work has assessed a broad range of toxicity tests, cell models and NP types and concentrations taking into account the inherent impact of NP properties and the effects of changes in experimental conditions using well-characterized NPs. The results of the studies have been used to generate recommendations for a suitable and robust testing strategy which can be applied to new medical NPs as they are developed.

Environmental fate of chlorinated bornanes estimated by theoretical descriptors.

Theoretical molecular descriptors have been calculated for 36 polychlorinated bornanes, the majority compound class of the insecticide Toxaphene. The results demonstrate that thermodynamic stability by the use of molecular structural energies can be used as a general parameter for persistence. Since these descriptors agree well with polychlorinated bornanes found in the environment, these compounds should be included as important indicator compounds in future trace analytical investigations of polychlorinated bornanes and also within experimental metabolism studies to investigate potential toxic metabolites. Reactivity descriptors such as electronaffinity, hardness, LUMO location and atomic charges may guide to potential chemical reactions like the dechlorination of polychlorinated bornanes in reductive environment. Further it is advised to use these descriptors and other new potential ones in combination with experimental degradation and toxicology studies to explore the relationship between molecular structure and biological effects of chlorobornanes.

Comparative molecular dynamics of mesophilic and psychrophilic protein homologues studied by 1.2 ns simulations.

It is well established that the dynamic motion of proteins plays an important functional role, and that the adaptation of a protein molecule to its environment requires optimization of internal non-covalent interactions and protein-solvent interactions. Serine proteinases in general, and trypsin in particular has been used as a model system in exploring possible structural features for cold adaptation. In this study, a 500 p.s. and a 1200 p.s. molecular dynamics (MD) simulation at 300 K of both anionic salmon trypsin and cationic bovine trypsin are analyzed in terms of molecular flexibility, internal non-covalent interactions and protein-solvent interactions. The present MD simulations do not indicate any increased flexibility of the cold adapted enzyme on an overall basis. However, the apparent higher flexibility and deformability of the active site of anionic salmon trypsin may lower the activation energy for ligand binding and for catalysis, and might be a reason for the increased binding affinity and catalytic efficiency compared to cationic bovine trypsin.

Comparative molecular dynamics simulation studies of salmon and bovine trypsins in aqueous solution.

The flexibility and conformational behaviour of salmon and bovine trypsins were modelled with a 300 ps molecular dynamics simulation in aqueous solution. Trajectories from both trypsins were analysed to eventually detect differences in mobility that could explain observed variations in stability and activity. The simulations were performed at 300 K with all the acidic groups deprotonated and the basic groups protonated. The radius of gyration, the overall r.m.s. deviation from the starting structure as a function of time, together with the r.m.s. deviation from the starting structures as a function of residue number, demonstrated that the simulations were stable and representative of the X-ray structures of both enzymes. Isotropic Debye-Waller factors were calculated from the fluctuations for main-chain atoms and were in good agreement with experimental values. The overall dynamic properties of the two enzymes were similar. Based on the present 300 ps molecular dynamics simulation, it cannot be concluded that either of the two enzymes is more 'flexible' than the other. However, there are clearly differences in mobility on a more detailed level and for particular regions.

Cold adaption of enzymes: structural comparison between salmon and bovine trypsins.

The crystal structure of an anionic form of salmon trypsin has been determined at 1.82 A resolution. We report the first structure of a trypsin from a phoikilothermic organism in a detailed comparison to mammalian trypsins in order to look for structural rationalizations for the cold-adaption features of salmon trypsin. This form of salmon trypsin (ST II) comprises 222 residues, and is homologous to bovine trypsin (BT) in about 65% of the primary structure. The tertiary structures are similar, with an overall displacement in main chain atomic positions between salmon trypsin and various crystal structures of bovine trypsin of about 0.8 A. Intramolecular hydrogen bonds and hydrophobic interactions are compared and discussed in order to estimate possible differences in molecular flexibility which might explain the higher catalytic efficiency and lower thermostability of salmon trypsin compared to bovine trypsin. No overall differences in intramolecular interactions are detected between the two structures, but there are differences in certain regions of the structures which may explain some of the observed differences in physical properties. The distribution of charged residues is different in the two trypsins, and the impact this might have on substrate affinity has been discussed.

Molecular structure and dynamics of the four 10-hydroxynortriptyline isomers.

The three-dimensional structures, molecular conformations, and electrostatic potentials of the R-E-, S-E-, R-Z-, and S-Z-isomers of 10-hydroxynortriptyline were examined by computer graphics, molecular mechanical energy calculations, and molecular dynamics simulations in vacuo and in aqueous solution. Molecular models of the isomers, based on the structure of nortriptyline, were refined by energy minimization and used as starting points in the simulations. R-E- and S-Z-10-hydroxynortriptyline formed intramolecular hydrogen bonds between the side-chain nitrogen atom and the hydroxyl group during the simulations in vacuo, and had the side chain folded over the ring system in the minimum energy conformations. Intramolecular hydrogen bonding was not observed for R-Z- and S-E-10-hydroxynortriptyline, which had extended side chains in the minimum energy conformations and stronger negative molecular electrostatic potentials around the hydroxyl group than the R-E- and S-Z-isomers.

Molecular structure and dynamics of tricyclic antidepressant drugs.

The molecular structure, electrostatic potentials and dynamics of imipramine, chlorimipramine, amitriptyline and nortriptyline were examined by computer graphics, molecular mechanical energy calculations and molecular dynamics simulations, using the AMBER all atom force field. Starting coordinates for amitriptyline and nortriptyline were generated by model building from the crystal structure of imipramine. The structures were refined by molecular mechanical energy minimization, and used as starting points for molecular dynamics simulations in vacuo and in aqueous solution. The simulations demonstrated considerable flexibility of the molecules, both in the side chain and in the ring system, where the angle between the phenyl rings varied between 90 degrees and 168 degrees. The most frequently observed conformations of imipramine, chlorimipramine and nortriptyline during the simulations had the side chain folded above one of the phenyl rings, while amitriptyline showed both folded and extended side chain conformations during the simulations. The results may provide increased understanding of the molecular recognition and specificity of tricyclic antidepressant drugs in interaction with neurotransmitter receptor molecules.