Cys mutants in functional regions of Sticholysin I clarify the participation of these residues in pore formation

Experimental evidence shows that the mechanism of pore formation by actinoporins is a multistep process, involving binding of the water-soluble monomer to the membrane and subsequent oligomerization on the membrane surface, leading to the formation of a functional pore. However, as for other eukaryotic pore-forming toxins, the molecular details of the mechanism of membrane insertion and oligomerization are not clear. In order to obtain further insight with regard to the structure-function relationship in sticholysins, we designed and produced three cysteine mutants of recombinant sticholysin I (rStI) in relevant functional regions for membrane interaction: StI E2C and StI F15C (in the N-terminal region) and StI R52C (in the membrane binding site). The conformational characterization derived from fluorescence and CD spectroscopic studies of StI E2C, StI F15C and StI R52C suggests that replacement of these residues by Cys in rStI did not noticeably change the conformation of the protein. The substitution by Cys of Arg(52) in the phosphocholine-binding site, provoked noticeable changes in rStI permeabilizing activity; however, the substitutions in the N-terminal region (Glu(2), Phe(15)) did not modify the toxin`s permeabilizing ability. The presence of a dimerized population stabilized by a disulfide bond in the StI E2C mutant showed higher pore-forming activity than when the protein is in the monomeric state, suggesting that sticholysins pre-ensembled at the N-terminal region could facilitate pore formation. (C) 2011 Elsevier Ltd. All rights reserved.

Application of solid sampling device for direct determination of chromium and nickel in lubricating oils by graphite furnace atomic absorption spectrometry

One method using a solid sampling device for the direct determination of Cr and Ni in fresh and used lubricating oils by graphite furnace atomic absorption spectrometry are proposed. The high organic content in the samples was minimized using a digestion step at 400 degrees C in combination with an oxidant mixture 1.0% (v v(-1)) HNO3+15% (v v(-1)) H2O2+0.1% (m v(-1)) Triton X-100 for the in situ digestion. The 3-field mode Zeeman-effect allowed the spectrometer calibration up to 5 ng of Cr and Ni. The quantification limits were 0.86 mu g g(-1) for Cr and 0.82 mg g(-1) for Ni, respectively. The analysis of reference materials showed no statistically significant difference between the recommended values and those obtained by the proposed methods.

Russian Arctic Ports : Problems with the development and management of tourist flows

The potential changes to the territory of the Russian Arctic open up unique possibilities for the development of tourism. More favourable transport opportunities along the Northern Sea Route (NSR) create opportunities for tourism development based on the utilisation of the extensive areas of sea shores and river basins. A major challenge for the Russian Arctic sea and river ports is their strong cargo transport orientation originated by natural resource extraction industries. A careful assessment of the prospects of current and future tourism development is presented here based on the development of regions located along the shores of the Arctic ocean (including Murmansk and Arkhangelsk oblast, Nenets Autonomous okrug (AO), Yamal-Nenets AO, Taymyr AO, Republic of Sakha, Chykotsky AO). An evaluation of the present development of tourism in maritime cities suggests that a considerable qualitative and quantitative increase of tourism activities organised by domestic tourism firms is made virtually impossible. There are several factors contributing to this. The previously established Soviet system of state support for the investments into the port facilities as well as the sea fleet were not effectively replaced by creation of new structures. The necessary investments for reconstruction could be contributed by the federal government but the priorities are not set towards the increased passenger transportation. Having in mind, increased environmental pressures in this highly sensitive area it is especially vital to establish a well-functioning monitoring and rescue system in the situation of ever increasing risks which come not only from the increased transports along the NSR, but also from the exploitation of the offshore oil and gas reserves in the Arctic seas. The capacity and knowledge established in Nordic countries (Norway, Finland) concerning cruise tourism should not be underestimated and the already functioning cooperation in Barents Region should expand towards this particular segment of the tourism industry. The current stage of economic development in Russia makes it clear that tourism development is not able to compete with the well-needed increase in the cargo transportation, which means that Russia’s fleet is going to be utilised by other industries. However, opening up this area to both local and international visitors could contribute to the economic prosperity of these remote areas and if carefully managed could sustain already existing maritime cities along the shores of the Arctic Ocean.