Energy migration in novel pH-triggered self-assembled beta-sheet ribbons

Energy migration between tryptophan residues has been experimentally demonstrated in self-assembled peptide tapes. Each peptide contains 11 amino acids with a Trp at position 6. The peptide self-assembly is pH-sensitive and forms amphiphilic tapes, which further stack in ribbons (double tapes) and fibrils in water depending on the concentration. Fluorescence spectra, quenching, and anisotropy experiments showed that when the pH is lowered from 9 to 2, the peptide self-assembly buries the tryptophan in a hydrophobic and restricted environment in the interior of stable ribbons as expected on the basis of the peptide design. These fluorescence data support directly and for the first time the presence of such ribbons which are characterized by a highly packed and stable hydrophobic interior. In common with Trp in many proteins, fluorescence lifetimes are nonexponential, but the average lifetime is shorter at low pH, possibly due to quenching with neighboring Phe residues. Unexpectedly, time-resolved fluorescence anisotropy does not change significantly with self-assembly when in water. In highly viscous sucrose-water mixtures, the anisotropy decay at low pH was largely unchanged compared to that in water, whereas at high pH, the anisotropy decay increased significantly. We concluded that depolarization at low pH was not due to rotational diffusion but mainly due to energy migration between adjacent tryptophan residues. This was supported by a master equation kinetic model of Trp-Trp energy migration, which showed that the simulated and experimental results are in good agreement, although on average only three Trp residues were visited before emission.     

NMR and UV studies of 3'-S-phosphorothiolate modified DNA in a DNA : RNA hybrid dodecamer duplex; implications for antisense drug design

High-resolution NMR spectroscopy has been used to establish the conformational consequences of the introduction of a single 3[prime or minute]-S-phosphorothiolate link in the DNA strand of a DNA : RNA hybrid. These systems are of interest as potential antisense therapeutic agents. Previous studies on similarly modified dinucleotides have shown that the conformation of the sugar to which the sulfur is attached shifts to the north (C(3[prime or minute])-endo/C(2[prime or minute])-exo). Comparisons made between NOESY cross-peak intensities, and coupling constants from PE-COSY spectra, for both non-modified and modified duplexes confirm that this conformational shift is also present in the double helical oligonucleotide system. In addition it is noted that in both the dinucleotides and the modified duplex, the conformation of the sugar ring 3[prime or minute] to the site of modification is also shifted to the north. That this pattern is observed in the small monomeric system as well as the larger double helix is suggestive of some pre-ordering of the sequences. The conclusion is supported by consideration of the (1)H chemical shifts of the heterocyclic bases near the site of the modification. The enhanced stability that these conformational changes should bring was confirmed by UV thermal melting studies. Subsequently a series of singly and doubly 3[prime or minute]-S-phosphorothiolate-modified duplexes were investigated by UV. The results are indicative of an additive effect of the modification with thermodynamic benefit being derived from alternate spacing of two modified linkers.     

Asymptotic stability in nonlinear elastic materials with dissipation

This paper is concerned with the effects of dissipation on classical solutions of the displacement and mixed initial-boundary value problems for a nonlinear elastic material. A proof of the asymptotic stability of the null solution in three dimensions is presented and the necessary assumptions are carefully discussed. The derivation of an analogous result for non-zero equilibrium solutions of the one-dimensional problem completes this work.     

Some static problems for elastic bodies with a crust

Summary This paper studies the diffusion of stresses in a linear isotropic elastic body coated with a thin elastic shell called the ?crust?. Two static problems are considered. The first is that of the semi-infinite half-space covered by a thin plate subject to a point load at the origin. In the second problem a solid elastic sphere enveloped by a thin spherical elastic shell with two opposite point loads at the poles is investigated. Solutions for the displacement within the thin crust are obtained in the two problems by means of the Hankel transform and the Legendre polynomial respectively.

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Sommario Questo lavoro studia la diffusione degli sforzi in un corpo elastico, lineare, isotropo rivestito da un guscio elastico sottile denominato la ?crosta?. Sono considerati due problemi elastostatici. Il primo è quello di un semispazio coperto da una piastra sottile soggetta and un carico concentrato nell'origine. Nel secondo problema si studia una sfera solida elastica avvolta da un guscio sottile elastico con due carichi concentrati opposti nei poli. Nei due problemi le soluzioni in termini di spostamenti entro la crosta si ottengono rispettivamente mediante la trasformata di Hankel e i polinomi di Legendre.

     

Polymer characterization by refractometry

In the use of refractometry for polymer characterization, it is shown that interpretation of data through use of a molar refractivity as defined by the Lorenz-Lorentz equation is more accurate than by use of the parachor. The disagreement between experimental and calculated values of the unit molar refractivity may be attributed to the arbitrary definition of the residue unit and the method of summation of group refractivities. In general, refractive index should depend on molecular weight and the conditions of polymerization, but this may depend on the dielectric properties and volume fraction of the domains comprising chain ends, initiator fragments, and free volume. The effect of polymer conformation on the molar refractivity of polyisoprene is found to be slight. For copolymer systems, there is a linear dependence of refractive index on weight fractional composition only where there is little order. The refractive index of a copolymer is not independent of the nature of the distribution of the residues along the chain.     

The role of twinning in the fatigue behaviour of α-titanium

Coarse grained (0·3 mm) -titanium has been fatigued in tension-compression about a zero mean load to 100 cycles/sec at normal temperature. Sections through fatigued specimens revealed that twin formation had occurred in grains completely constrained by surrounding material during cyclic loading and that fatigue damage was associated with some of these twins. A micro-beam X-ray and single face sterographic analysis technique combined with metallographic observations of the specimens revealed that {10¯12}, {11¯21} and {11¯22} twins had formed and that the internal fatigue damage was preferentially formed at the twin-matrix interface of the {11¯21} twins.The authors wish to thank Professor G. V. Raynor, F.R.S., for the provision of laboratory facilities. This research has been sponsored in part by the Air Force Materials Laboratory, Wright-Patterson Air Force Base, Dayton, Ohio U.S.A. through the European Office of Aerospace Research under Contract No. F61052.67.C.0007.     

Precise mass measurement of a double-stranded 500 base-pair (309 kDa) polymerase chain reaction product by negative ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS) has been used to determine the mass of a double-stranded 500 base-pair (bp) polymerase chain reaction (PCR) product with an average theoretical mass of the blunt-ended (i.e. unadenylated) species of 308 859.35 Da. The PCR product was generated from the linearized bacteriophage Lambda genome which is a double-stranded template. Utilization of ethanol precipitation in tandem with a rapid microdialysis step to purify and desalt the PCR product was crucial to obtain a precise mass measurement. The PCR product (0.8 pmol/μL) was electrosprayed from a solution containing 75% acetonitrile, 25 mM piperidine, and 25 mM imidazole and was infused at a rate of 200 nL/min. The average molecular mass and the corresponding precision were determined using the charge-states ranging from 172 to 235 net negative charges. The experimental mass and corresponding precision (reported as the 95% confidence interval of the mean) was 309 406 +/- 27 Da (87 ppm). The mass accuracy was compromised due to the fact that the PCR generates multiple products when using Taq polymerase due to the non-template directed 3'-adenylation. This results in a mixture of three PCR products with nearly identical mass (i.e. blunt-ended, mono-adenylated and di-adenylated) with unknown relative abundances that were not resolved in the spectrum. Thus, the experimental mass will be a weighted average of the three species which, under our experimental conditions, reflects a nearly equal concentration of the mono- and di-adenylated species. This report demonstrates that precise mass measurements of PCR products up to 309 kDa (500 bp) can be routinely obtained by ESI-FTICR requiring low femtomole amounts. Copyright 1999 John Wiley & Sons, Ltd.     

Investigation of the quantitative properties of the quadrupole orthogonal acceleration time-of-flight mass spectrometer with electrospray ionisation using 3,4-methylenedioxymethamphetamine

This paper describes the investigation of the potential of a quadrupole orthogonal acceleration time-of-flight mass spectrometer (Q-TOF) equipped with an atmospheric pressure ionisation interface for quantitative measurements of small molecules separated by reversed phase liquid chromatography. To this end, the detection limits and linear dynamic range in particular were studied in an LC/MS/MS experiment using 3,4-methylenedioxymethamphetamine standards and 3,4-methylenedioxyethylamphetamine for internal standardisation. In a second phase, the experiment was repeated with real biological extracts (whole blood, serum, and vitreous humour). A calibration for 3,4-methylenedioxymethamphetamine and its metabolite 3,4-methylenedioxyamphetamine was prepared in each of these matrices again using 3,4-methylenedioxyethylamphetamine as internal standard. The resulting quantitative data were compared with those obtained by liquid chromatography with fluorescence detection for the same extracts. The Q-TOF results revealed excellent sensitivity and a linear dynamic range of nearly four decades (2-10 000 pg on-column, r(2) = 0.9998, 1/x weighting). Furthermore, all the calibration curves prepared in biological material were superimposable, LC/MS/MS and LC-fluorescence, and the quantitative results for actual samples compared very favourably. It was concluded that the Q-TOF achieves a linear dynamic range for quantitative LC/MS/MS work exceeding that of fluorescence detection and at much better absolute sensitivity. Copyright 1999 John Wiley & Sons, Ltd.     

Some stability results in the linear theory of anisotropic elastodynamics

Conditions are established to ensure continuous data dependence for classical solutions of certain initial-boundary value problems in linearised anisotropic elastodynamics. Two uniqueness theorems are also presented.

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Zusammenfassung Bedingungen werden definiert, damit die klassischen Lösungen gewisser Probleme des Anfangsgrenzwertes auf dem Gebiet der linearen anisotropischen elastodynamik stetig und direkt von Daten abhängen. In diesem Zusammenhang werden dann zwei Eindeutigkeitssätze erklärt.