Peptide internal motions on nanosecond time scale derived from direct fitting of (13)C and (15)N NMR spectral density functions

NMR relaxation-derived spectral densities provide information on molecular and internal motions occurring on the picosecond to nanosecond time scales. Using (13)C and (15)N NMR relaxation parameters [T(1), T(2), and NOE] acquired at four Larmor frequencies (for (13)C: 62.5, 125, 150, and 200 MHz), spectral densities J(0), J(omega(C)), J(omega(H)), J(omega(H) + omega(C)), J(omega(H) - omega(C)), J(omega(N)), J(omega(H) + omega(N)), and J(omega(H) - omega(N)) were derived as a function of frequency for (15)NH, (13)C(alpha)H, and (13)C(beta)H(3) groups of an alanine residue in an alpha-helix-forming peptide. This extensive relaxation data set has allowed derivation of highly defined (13)C and (15)N spectral density maps. Using Monte Carlo minimization, these maps were fit to a spectral density function of three Lorentzian terms having six motional parameters: tau(0), tau(1), tau(2), c(0), c(1), and c(2), where tau(0), tau(1) and tau(2) are correlation times for overall tumbling and for slower and faster internal motions, and c(0), c(1), and c(2) are their weighting coefficients. Analysis of the high-frequency portion of these maps was particularly informative, especially when deriving motional parameters of the side-chain methyl group for which the order parameter is very small and overall tumbling motions do not dominate the spectral density function. Overall correlation times, tau(0), are found to be in nanosecond range, consistent with values determined using the Lipari-Szabo model-free approach. Internal motional correlation times range from picoseconds for methyl group rotation to nanoseconds for backbone N-H, C(alpha)-H, and C(alpha)-C(beta) bond motions. General application of this approach will allow greater insight into the internal motions in peptides and proteins.     

Designing protein beta-sheet surfaces by Z-score optimization

Studies of lattice models of proteins have suggested that the appropriate energy expression for protein design may include nonthermodynamic terms to accommodate negative design concerns. One method, developed in lattice model studies, maximizes a quantity known as the " Z-score," which compares the lowest energy sequence whose ground state structure is the target structure to an ensemble of random sequences. Here we show that, in certain circumstances, the technique can be applied to real proteins. The resulting energy expression is used to design the beta-sheet surfaces of two real proteins. We find experimentally that the designed proteins are stable and well folded, and in one case is even more thermostable than the wild type.     

Experimental oscillator strengths of Zn II lines of astrophysical interest

Relative transition probabilities for 18 lines of Zn II (11 measured for the first time) were determined from the emission-line intensities in a laser-produced plasma. The experiment was carried out using Cd–Zn alloy with a Zn content under 10% in order to obtain optically thin plasma. Oscillator strengths were placed on an absolute scale by using experimental lifetimes, line-strength sum rules and Boltzmann plot. A comparison has been conducted between present experimental results, the data available and new calculations reported in this work, as well as a study of the plasma conditions.     

Phase separation of polymer‐functionalized SWNTs within a PMMA/polystyrene blend

Phase separation of polystyrene (PS) and poly(methyl methacrylate) (PMMA) blends was used as a means to segregate PS‐ or PMMA‐functionalized single‐walled carbon nanotubes (SWNTs) in thin films. Dilute solutions (5 wt % in THF) of 1:1 PS/PMMA blends containing the functionalized nanotubes were spin cast and annealed at 180 °C for 12 h. Two different polymer molecular weights were used (M_n = 8000 or M_n = 22,000), and were of approximately equivalent molecular weight to those attached to the surface of the nanotubes. Nanotube functionalization was accomplished using the Cu(I)‐catalyzed [3 + 2] Huisgen cycloaddition, in which alkyne‐decorated nanotubes were coupled with azide‐terminated polymers, resulting in polymer‐SWNT conjugates that were soluble in THF. Characterization of the annealed films by scanning Raman spectroscopy, which utilized the unique Raman fingerprint of carbon nanotubes, enabled accurate mapping of the functionalized SWNTs within the films relative to the two phase‐separated polymers. It was found that nanotube localization within the phase‐separated polymer films was influenced by the type of polymer attached to the nanotube surface, as well as its molecular weight. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 450–458, 2009     

Adult-child differences in acoustic cue weighting are influenced by segmental context: children are not always perceptually biased toward transitions

It has been proposed that young children may have a perceptual preference for transitional cues [Nittrouer, S. (2002). J. Acoust. Soc. Am. 112, 711-719]. According to this proposal, this preference can manifest itself either as heavier weighting of transitional cues by children than by adults, or as heavier weighting of transitional cues than of other, more static, cues by children. This study tested this hypothesis by examining adults' and children's cue weighting for the contrasts /saI/-/integral of aI/, /de/-/be/, /ta/-/da/, and /ti/-/di/. Children were found to weight transitions more heavily than did adults for the fricative contrast /saI/-/integral aI/, and were found to weight transitional cues more heavily than nontransitional cues for the voice-onset-time contrast /ta/-/da/. However, these two patterns of cue weighting were not found to hold for the contrasts /de/-/be/ and /ti/-/di/. Consistent with several studies in the literature, results suggest that children do not always show a bias towards vowel-formant transitions, but that cue weighting can differ according to segmental context, and possibly the physical distinctiveness of available acoustic cues.     

An integrated enzyme-linked immunosorbent assay system with an organic light-emitting diode and a charge-coupled device for fluorescence detection

A fluorescence detection system for a microfluidic device using an organic light-emitting diode (OLED) as the excitation light source and a charge-coupled device (CCD) as the photo detector was developed. The OLED was fabricated on a glass plate by photolithography and a vacuum deposition technique. The OLED produced a green luminescence with a peak emission at 512 nm and a half bandwidth of 55 nm. The maximum external quantum efficiency of the OLED was 7.2%. The emission intensity of the OLED at 10 mA/cm(2) was 13 μW (1.7 mW/cm(2)). The fluorescence detection system consisted of the OLED device, two band-pass filters, a five microchannel poly(dimethylsiloxane) (PDMS) microfluidic device and a linear CCD. The fluorescence detection system was successfully used in a flow-based enzyme-linked immunosorbent assay on a PDMS microfluidic device for the rapid determination of immunoglobulin A (IgA), a marker for human stress. The detection limit (S/N=3) for IgA was 16.5 ng/mL, and the sensitivity was sufficient for evaluating stress. Compared with the conventional 96-well microtiter plate assay, the analysis time and the amounts of reagent and sample solutions could all be reduced.     

Crystallization Elution Fractionation and Thermal Gradient Interaction Chromatography. Techniques Comparison

Summary: The chemical composition distribution has been shown to be the most critical and discriminating parameter in understanding the performance of industrial polyolefins with non homogeneous comonomer incorporation. The chemical composition distribution is being analyzed by well known techniques such as temperature rising elution fractionation, TREF, crystallization analysis fractionation, CRYSTAF and crystallization elution fractionation, CEF. These techniques separate according to crystallizability and provide a powerful and predictable separation of components based on the presence of branches, irregularities or tacticity differences, independently of the molar mass. TREF, CRYSTAF and CEF can not be used, however, for the separation of more amorphous resins, and may not always provide the best solution for complex multi-component resins due to the existence of some co-crystallization. The application of high temperature interactive HPLC to polyolefins opened a new route to characterize these types of polymers. The use of solvent gradient HPLC for separation of polyethylene and polypropylene and the developments in HPLC on carbon based columns extended further the application of high temperature HPLC in polyolefins. A new approach has been developed recently using the carbon based column but replacing solvent gradient by a thermal gradient which facilitates the analysis of polyethylene copolymers and provides a powerful tool for the analysis of elastomers. Thermal gradient interaction chromatography (TGIC) is being compared with TREF and CEF with the analysis of model samples. The advantages/disadvantages of each technique are being investigated and discussed. The combination of TGIC and TREF/CEF provides an extended range of separation of polyolefins.     

Fabrication and performance of polymer–nanocomposite anti-reflective thin films deposited by RIR-MAPLE

Design of polymer anti-reflective (AR) optical coatings for plastic substrates is challenging because polymers exhibit a relatively narrow range of refractive indices. Here, we report synthesis of a four-layer AR stack using hybrid polymer:nanoparticle materials deposited by resonant infrared matrix-assisted pulsed laser evaporation. An Er:YAG laser ablated frozen solutions of a high-index composite containing TiO₂ nanoparticles and poly(methyl-methacrylate) (PMMA), alternating with a layer of PMMA. The optimized AR coatings, with thicknesses calculated using commercial software, yielded a coating for polycarbonate with transmission over 97 %, scattering <3 %, and a reflection coefficient below 0.5 % across the visible range, with a much smaller number of layers than would be predicted by a standard thin film calculation. The TiO₂ nanoparticles contribute more to the enhanced refractive index of the high-index layers than can be accounted for by an effective medium model of the nanocomposite.     

BIPHASIC PHOTOCHEMISTRY: THE FIRST ACTINOMETRIC REACTION ON A SOLID SUPPORT

Abstract Measurements were performed to determine the action spectrum and dose dependence for photoreactivation of E. coli B_s-1 cells after γ-irradiation. The similarities between photoreactivation after UV- and after γ-irradiation in action spectra, kinetics, and other characteristics indicate that the increased survival of γ-irradiated cells after illumination with photoreactivating light is the result of true photoenzymatic repair.