Partially reversible adsorption of annexin A1 on POPC/POPS bilayers investigated by QCM measurements, SFM, and DMC simulations

The kinetics of annexin A1 binding to solid-supported lipid bilayers consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS; 4:1) has been investigated as a function of the calcium ion concentration in the bulk phase. Quartz crystal microbalance measurements in conjunction with scanning force microscopy, fluorescence microscopy, and computer simulations indicate that at a given Ca2+ concentration annexin A1 adsorbs irreversibly on membrane domains enriched in POPS. By contrast, annexin A1 adsorbs reversibly on the POPC-enriched phase, which is composed of single POPS molecules embedded within a POPC matrix. The overall area occupied by the POPS-enriched phase is controlled by the CaCl2 concentration. Monte Carlo simulations suggest that the area of the POPS-enriched phase increases by a factor of 7 when the Ca2+ concentration is changed from 0.01 to 1 mM.     

Unusually strong positive cooperativity in binding of peptides to latent membrane protein-1 DNA fragments of the Epstein-Barr viral gene

The DNA-binding preferences of two oligopeptide amides, (His-Pro-Arg-Lys)(3)NH(2) (HR-12) and (Ser-Pro-Arg-Lys)(3)NH(2) (SP-12), have been examined by quantitative DNase I footprinting studies. Two different DNA fragments were investigated: a pair of 5'-(32)P-labeled duplexes from pBR322 with one or other of the complementary strands labeled and a corresponding pair of 5'-(32)P-labeled duplexes representing fragments of the latent membrane protein (LMP-1) gene from a pathogenic Epstein-Barr virus variant derived from nasopharyngeal carcinoma. The major objective was to examine molecular recognition and cooperative features associated with sequence-selective binding of synthetic peptides to the LMP-1 fragments. At various binding sites on the pBR322 fragments, Hill coefficients (n(H)) ranging from 1.9-2.2 were observed; these results indicate modest positive cooperativity between binding sites for both peptides. By contrast, unusually high values of n(H), ranging from 4.0-9.3, were observed at various binding sites on the LMP-1 fragments. Allosteric models can be constructed to interpret the observed cooperative interactions between different DNA recognition sites in the LMP-1 gene upon binding of the peptide ligands. It is noteworthy that these models feature a novel network of cooperativity interconnecting multiple DNA allosteric sites. The evidence of sequence selectivity and strong cooperativity discovered in this work may prove to be a general feature of peptide interactions with some nucleic acids.     

Paraelectric thin films by nanoscale engineering of epitaxy and planar anisotropy for microwave phase shifter applications

Epitaxial and (110) oriented paraelectric thin films of Ba0.60Sro.40TiO3 were grown on (100) oriented NdGaO3 orthorhombic substrates, and the nonlinear dielectric properties were studied at 10 GHz along selected in-plane crystallographic directions in the film thickness range of 25-1200 nm. The measured dielectric properties show strong residual strain and in-plane directional dependence. For instance, the in-plane relative permittivity is found to vary from as much as 500 to 150 along [110] and [001], respectively, in the 600 nm film. Tunability was found to vary from as much as 54% to 20% in all films and directions. In a given film, the best tunability is observed along the compressed axis in a mixed strain state, 54% along [110] in the 600 nm film. It is shown that, by nanoscale manipulation of epitaxy and planar anisotropy, the return loss and phase shift in a paraelectric can be tuned over a rather wide range. The approach presented herein opens avenues for obtaining various degrees of phase shift on the same film, enabling one with an additional degree of freedom in device design and fabrication as well as multifunctionality.     

Thermal cooling using low-temperature waste heat: a cost-effective way for industrial companies to improve energy efficiency?

As a typical cross-cutting technology, cooling and refrigeration equipment is used for a variety of industrial applications. While cooling is often provided by electric compression cooling systems, thermal cooling systems powered by low-temperature waste heat could improve energy efficiency and promise a technical saving potential corresponding to 0.5?% of the total electricity demand in the German industry. In this paper, we investigate the current and future cost-effectiveness of thermal cooling systems for industrial companies. Our focus is on single-stage, closed absorption and adsorption cooling systems with cooling powers between 40 and 100?kW, which use low-temperature waste heat at temperature levels between 70?°C and 85?°C. We analyse the current and future cost-effectiveness of these alternative cooling systems using annual cooling costs (annuities) and payback times. For a forecast until 2015, we apply the concept of experience curves, identifying learning rates of 14?% (absorption machines) and 17?% (adsorption machines) by an expert survey of the German market. The results indicate that thermal cooling systems are currently only cost-effective under optimistic assumptions (full-time operation, high electricity prices) when compared to electric compression cooling systems. Nevertheless, the cost and efficiency improvements expected for this still young technology mean that thermal cooling systems could be more cost-effective in the future. However, depending on future electricity prices, a high number of operating hours is still crucial to achieve payback times substantially below 4?years which are usually required for energy efficiency measures to be widely adopted in the industry.     

Detection of low-abundance protein phosphorylation by selective (18)o labeling and mass spectrometry

Reversible phosphorylation regulates the majority of intracellular networking and pathways. The study of this widely explored post-translational modification is usually challenged by low stoichiometric levels of modification. Many approaches have been developed to overcome this problem and to achieve rigorous characterization of protein phosphorylation. We describe a method for enhanced detection of low-abundance protein phosphorylation that uses selective introduction of (18)O label into phosphorylation sites with H(2)(18)O and mass spectrometric detection. The method was applied to introduce (18)O label into bacterially expressed Aurora A kinase phosphorylation sites and resulted in the representation of phosphorylated peptides as doublets or triplets according to the number of phosphate groups. A total of 28 phosphopeptides were observed by this method.     

Rapid identification of long-chain polyunsaturated fatty acids in a marine extract by HPLC-MS using data-dependent acquisition

The collision-induced dissociation (CID) of a range of deprotonated fatty acid standards was studied using linear ion trap mass spectrometry. Neutral losses of 78, 98, and 136 Da were consistently observed for fatty acids with five or more double bonds. Comparison of the MS/MS spectra of docosahexaenoic acid (DHA) and universally (13)C-labeled DHA allowed the molecular formulas for these neutral losses to be determined as C(6)H(6), C(5)H(6)O(2), and C(8)H(8)O(2). Knowledge of fatty acid fragmentation processes was then applied to identify fatty acids from a sea anemone, Aiptasia pulchella, and dinoflagellate symbiont, Symbiodinium sp. extract. Using HPLC-MS, fatty acids were separated and analyzed by tandem mass spectrometry in data-dependent acquisition mode. Neutral loss chromatograms for 78, 98, and 136 Da allowed the identification of long-chain fatty acids with five or more double bonds. On the basis of precursor ion m/z ratios, chain length and degree of unsaturation for these fatty acids were determined. The application of this technique to an Aiptasia sp.-Symbiodinium sp. lipid extract enabled the identification of the unusual, long-chain fatty acids 24:6, 26:6, 26:7, 28:7, and 28:8 during a single 40 min HPLC-MS analysis.     

Multiband sensor using thick holographic gratings for sulfur detection by laser-induced breakdown spectroscopy

Detection of sulfur by optical emission spectroscopy generally presents some difficulties because the strongest lines are in the vacuum UV below 185 nm and therefore are readily absorbed by oxygen molecules in air. A novel concept for a low-cost and efficient system to detect sulfur using near-IR bands by laser-induced breakdown spectroscopy is here proposed. This concept is based on customized thick holographic gratings as spectral filtering elements. The signal integration and the temporal synchronization are performed using built-in custom electronics that amplify and integrate or trigger photodiode output signals. In this work, we use the near-IR lines at 921.287 nm and a background reference at 900 nm. Preliminary results show a limit of detection comparable to that of a conventional high-end system.     

Critical assessment of UO2 classical potentials for thermal conductivity calculations

This article reviews the thermal transport properties as predicted by 26 classical interatomic potentials for uranium dioxide, an important nuclear fuel material, determined using a lattice dynamics-based method. The calculations reveal structural instabilities for multiple potentials, as well as the presence of lower energy structures even for potentials in which the fluorite structure is stable. Both rigid atom and shell model potentials are considered, and general trends in their representation of the thermal conductivity are identified. Reviewed classical potentials predict thermal conductivity in the range of ~5–22 W/mK, compared to the experimental value of 8.9 W/mK. The quality of the anharmonicity correction that is based on the correlation between thermal expansion and thermal conductivity is investigated, and it found to generally improve thermal conductivities results.