Carbon nanotubes in macrophages: imaging and chemical analysis by X-ray fluorescence microscopy

X-ray fluorescence microscopy (microXRF) is applied for the first time to study macrophages exposed to unpurified and purified single-walled (SW) and multiwalled (MW) carbon nanotubes (CNT). Investigating chemical elemental distributions allows one to (i) image nanotube localization within a cell and (ii) detect chemical modification of the cell after CNT internalization. An excess of calcium is detected for cells exposed to unpurified SWCNT and MWCNT and related toxicological assays are discussed.     

Fold and flexibility: what can proteins' mechanical properties tell us about their folding nucleus?

The determination of a protein''s folding nucleus, i.e. a set of native contacts playing an important role during its folding process, remains an elusive yet essential problem in biochemistry. In this work, we investigate the mechanical properties of 70 protein structures belonging to 14 protein families presenting various folds using coarse-grain Brownian dynamics simulations. The resulting rigidity profiles combined with multiple sequence alignments show that a limited set of rigid residues, which we call the consensus nucleus, occupy conserved positions along the protein sequence. These residues'' side chains form a tight interaction network within the protein''s core, thus making our consensus nuclei potential folding nuclei. A review of experimental and theoretical literature shows that most (above 80%) of these residues were indeed identified as folding nucleus member in earlier studies.     

The influence of clearance on friction, lubrication and squeaking in large diameter metal-on-metal hip replacements

Large diameter metal-on-metal bearings (MOM) are becoming increasingly popular, addressing the needs of young and more active patients. Clinical data has shown excellent short-to-mid-term results, though incidences of transient squeaking have been noted between implantation and up to 2 years post-operative. Geometric design features, such as clearance, have been significant in influencing the performance of the bearings. Sets of MOM bearings with different clearances were investigated in this study using a hip friction simulator to examine the influence of clearance on friction, lubrication and squeaking. The friction factor was found to be highest in the largest clearance bearings under all test conditions. The incidence of squeaking was also highest in the large clearance bearings, with all bearings in this group squeaking throughout the study. A very low incidence of squeaking was observed in the other two clearance groups. The measured lubricating film was found to be lowest in the large clearance bearings. This study suggests that increasing the bearing clearance results in reduced lubricant film thickness, increased friction and an increased incidence of squeaking.     

Surface-immobilized peptide aptamers as probe molecules for protein detection

We demonstrate the use of surface-immobilized, oriented peptide aptamers for the detection of specific target proteins from complex biological solutions. These peptide aptamers are target-specific peptides expressed within a protein scaffold engineered from the human protease inhibitor stefin A. The scaffold provides stability to the inserted peptides and increases their binding affinity owing to the resulting three-dimensional constraints. A unique cysteine residue was introduced into the protein scaffold to allow orientation-specific surface immobilization of the peptide aptamer and to ensure exposure of the binding site to the target solution. Using dual-polarization interferometry, we demonstrate a strong relationship between binding affinity and aptamer orientation and determine the affinity constant KD for the interaction between an oriented peptide aptamer ST(cys+)_(pep9) and the target protein CDK2. Further, we demonstrate the high selectivity of the peptide aptamer STM_(pep9) by exposing surface-immobilized ST(cys+)_(pep9) to a complex biological solution containing small concentrations of the target protein CDK2.     

Characterization and leachability of electric arc furnace dust made from remelting of stainless steel

Electric arc furnace dust (EAFD) is a toxic waste product made in the remelting of scrap steel. The results of a Toxicity Characteristic Leaching Procedure (TCLP) conducted on a sample of EAFD originating from the remelting of stainless steel scrap showed that the total Cr and Cr (VI) liquor concentrations (9.7 and 6.1 mg/L, respectively) exceeded the Toxicity Characteristic Regulatory Level (TCRL). The EAFD showed a complex heterogeneous mineralogy with spinel minerals group predominance. A sequential extractions method has permitted the determination of the amount of available metals (potentially mobile component) from the EAFD as follows: Cr (3%), Ni (6%), Pb (49%) and Zn (40%). Solubility controls on Cr, Pb, Zn and Ni were identified in the EAFD. This means that the Cr, Pb, Zn and Ni concentrations in solution were controlled by the solubility of some phases from EAFD. The concentrations of Ni and Zn, which are metals not regulated by TCRL were below 0.41 and 1.3 mg/L, respectively. The solubility control on Pb was sufficient to decrease its concentration (<0.24 mg/L) to a level below the TCRL. However, the control on Cr was not sufficient to decrease its concentration (between 117 and 331 mg/L) to below the TCRL.     

Dynamics of killer T cell inflation in viral infections

Upon acute viral infection, a typical cytotoxic T lymphocyte (CTL) response is characterized by a phase of expansion and contraction after which it settles at a relatively stable memory level. Recently, experimental data from mice infected with murine cytomegalovirus (MCMV) showed different and unusual dynamics. After acute infection had resolved, some antigen specific CTL started to expand over time despite the fact that no replicative virus was detectable. This phenomenon has been termed as "CTL memory inflation". In order to examine the dynamics of this system further, we developed a mathematical model analysing the impact of innate and adaptive immune responses. According to this model, a potentially important contributor to CTL inflation is competition between the specific CTL response and an innate natural killer (NK) cell response. Inflation occurs most readily if the NK cell response is more efficient than the CTL at reducing virus load during acute infection, but thereafter maintains a chronic virus load which is sufficient to induce CTL proliferation. The model further suggests that weaker NK cell mediated protection can correlate with more pronounced CTL inflation dynamics over time. We present experimental data from mice infected with MCMV which are consistent with the theoretical predictions. This model provides valuable information and may help to explain the inflation of CMV specific CD8+T cells seen in humans as they age.     

Mechanically responding nanovalves based on polyelectrolyte multilayers

The alternate deposition of exponentially and linearly growing polyelectrolyte multilayers leads to the formation of multicompartment films. In this study, a new system consisting in nanometer-sized multilayer barriers deposited on or between multilayer compartments was designed to respond to mechanical stimuli and to act as nanovalves. The diffusion of polyelectrolytes through the barrier from one compartment to another can be switched on/off by tuning the mechanical stretching and thereby opening or closing nanopores in the barrier. This work represents a first step toward the design of chemically or biologically active films responding to mechanical stresses.