Solid‐state NMR Study of the YadA Membrane‐Anchor Domain in the Bacterial Outer Membrane

MAS‐NMR was used to study the structure and dynamics at ambient temperatures of the membrane‐anchor domain of YadA (YadA‐M) in a pellet of the outer membrane of E. coli in which it was expressed. YadA is an adhesin from the pathogen Yersinia enterocolitica that is involved in interactions with the host cell, and it is a model protein for studying the autotransport process. Existing assignments were sucessfully transferred to a large part of the YadA‐M protein in the E. coli lipid environment by using ¹³C‐¹³C DARR and PDSD spectra at different mixing times. The chemical shifts in most regions of YadA‐M are unchanged relative to those in microcrystalline YadA‐M preparations from which a structure has previously been solved, including the ASSA region that is proposed to be involved in transition‐state hairpin formation for transport of the soluble domain. Comparisons of the dynamics between the microcrystalline and membrane‐embedded samples indicate greater flexibility of the ASSA region in the outer‐membrane preparation at physiological temperatures. This study will pave the way towards MAS‐NMR structure determination of membrane proteins, and a better understanding of functionally important dynamic residues in native membrane environments.     

Interaction forces between silica surfaces in aqueous solutions of cationic polymeric flocculants: effect of polymer charge

Three cationic polymers with molecular weights and charge density of 3.0 x 10(5) g/mol and 10% (D 6010), 1.1 x 10(5) g/mol and 40% (D6040), and 1.2 x 10(5) g/mol and 100% (D6099) were investigated in aqueous NaCl solutions in the presence of silica. The atomic force microscope (AFM) colloidal probe technique was used to determine silica interparticle interaction forces, which were compared to macroscopic information on the strength of interactions such as compressive yield stress measurements. It was found that in 30 mM NaCl solution the 10% charged polymer produced steric repulsion upon approach and long-range adhesion with multiple pull off events upon retraction at the optimum flocculation concentration. This suggests that the polymer was adsorbed in a conformation where segments extend from the surface, resulting in bridging flocculation. The 40 and 100% charged polymers produced attraction upon approach and strong adhesion with snap out from contact upon separation at optimum polymer dosages. This suggests that these polymers are adsorbed with flat conformations and is typical of charge neutralization or patch attraction. The attractions for 40 and 100% charged polymers measured with the AFM are significantly larger than for the 10% charged polymer. The polymer dose that produced the optimum flocculation and the maximum compressive yield stress typically corresponded to the polymer concentration that produced the maximum adhesion for each polymer. It was found that the magnitude of the adhesive force was more significant in determining the compressive yield stresses of the silica particle sediments than the aggregate size and structure.     

A study of beryllium-hydrogencomplexes in silicon

Single-crystal silicon, containing the impurity beryllium, has been doped with hydrogen, using two different techniques. Some samples were implanted with 3 MeV protons as a hydrogen source, and others were heated in a hydrogen atmosphere up to temperatures of 1000°C. Infrared absorption spectra of these samples at low temperatures have revealed two new series of absorptions identified as acceptor levels 91 meV and about 73 meV above the valence band. Quenching and annealing studies indicate that these absorptions are due to beryllium-hydrogen pairs, a direct analog to the beryllium-lithium pairs. Also a new series of absorptions has been found in samples which have a very high beryllium concentration. These lines at 298, 324 and 345 cm^?1 disappear after short anneals at elevated temperatures and are believed to be an acceptor level associated with a more complex beryllium center.     

Stimulation of healing of varicose ulcers by ultrasound

The local application of theraputic ultrasound has been shown, in a controlled trial, to stimulate the healing of chronic varicose ulcers. The change in ulcer size by the end of the period of treatment was noted. It was found that healing of insonated ulcers was significantly more marked than that exhibited by the controls.     

Assignments of visible absorption of s-tetrazine vapor

A detailed vibrational analysis of the 5515 Å absorption of s-tetrazine (tetra-azabenzene) vapor is attempted. More than 80% of the total intensity of the system can be accounted for by band assignments to a single allowed electronic transition, ¹B_3u-¹A_g. The only progression-forming vibration in absorption in ν_6a. Three progressions in ν′_6a account for all intense cold bands. Isotopic shifts of bands lead to convincing assignments of the two excited-state vibrations upon which ν_6a progressions are based, namely ν_6b and ν_8a. It is shown that in all probability the antisymmetric mode ν_6b is prominent because of vibrational (Fermi) resonance rather than because of vibronic interactions.     

Bi-modal hetero-aggregation rate response to particle dosage

The rate of flocculation of cationic polystyrene latex (PSL) particles by smaller, anionic PSL particles has been measured using a low-angle static light scattering technique. The rate of aggregate growth has been investigated as a function of particle size ratio and relative concentration of each particle species (for a constant dose of cationic particles). Contrary to many previous reports, two peaks in the flocculation rate were observed as a function of dose. It is speculated that the peak observed at the lower particle concentration coincides with the dose yielding maximum constant collision efficiency in the steady-state regime, a condition which is attained only after complete adsorption of the smaller particles onto the larger particle species. The peak at the higher particle concentration is believed to be related to the maximum collision rate constant upon reaching the steady-state regime, the value of which corresponds to maximum degree of aggregation and therefore the maximum mean collision efficiency prior to reaching this condition. From classical collision kinetics, the rate of aggregate growth may be represented as being proportional to the product of the collision rate constant and collision efficiency at any given time. Given then that the maximum value of these two variables coincides with different particle concentrations, the product of the response of each to particle dosage can in some cases yield a net bi-modal aggregation rate response to particle dosage.     

The resonance Raman spectrum of ultramarine blue

The resonance Raman spectrum of ultramarine blue has been observed by the use of 457.9, 488.0, 514.5, 568.2 and 647.1 nm excitation. With 457.9 nm excitation three progressions are observed; comparison of the results with earlier data from sulphur-doped alkali halide crystals indicates that ultramarine blue contains the S₂^? as well as the S₃^? anion.     

Stimulant sensitive flocculation and consolidation for improved solid/liquid separation

A novel method of flocculation resulting in both rapid sedimentation and low sediment moisture is described. It relies on changing the inter-particle forces from repulsive to attractive (aggregation and fast settling results) and then back to repulsive (densification of sediment then occurs). The change in inter-particle force is controlled by a stimulus such as pH or temperature. The technique is demonstrated without polymer using the isoelectric point and pH as the stimulus. The polyelectrolyte chitosan produces faster sedimentation and clearer supernatants as a pH sensitive flocculant. Methylcellulose is an effective temperature sensitive flocculant. The sediment bed volumes can be reduced by between 10 and 45% depending upon the conditions which, as yet, have not been fully optimized.