Determination of the rate of rapid lipid transfer induced by poly(ethylene glycol) using the SLM Fourier transform phase and modulation spectrofluorometer

Rate constants were determined for the transfer of the fluorescent lipid probe 1-palmitoyl-2-[[2-[4-(6-phenyl-trans-1,3,5-hexatrienyl)phenyl]ethyl] oxy]carbonyl]-3-sn-phosphatidylcholine (DPHpPC) between large, unilamellar extrusion vesicles composed either of dipalmitoyl phosphatidylcholine (DPPC) or of DPPC mixed with a small amount (0.5 mol%) of lyso phosphatidylcholine (Lyso PC). Transfer of the lipid probe in the presence of varying concentrations of poly(ethylene glycol) (PEG) was monitored using the SLM 48000-MHF Multi-Harmonic Fourier Transform phase and modulation spectrofluorometer to collect multifrequency phase and modulation fluorescence data sets on a subsecond time scale. The unique ability of this instrument to yield accurate fluorescence lifetime data on this time scale allowed transfer to be detected in terms of a time-dependent change in the fluorescent lifetime distribution associated with the lipid-like DPHpPC probe. This probe demonstrates two short fluoresence decay times (ca. 1.1–1.4 and 4.3–4.8 ns) in a probe-rich environment but a single long lifetime (ca. 7 ns) in a probe-poor environment. A simple two-state model for initial lipid transfer was used to analyze the multifrequency data sets collected over a 4-s time frame to obtain the time rate of change of the concentrations of donor and acceptor probe populations following rapid mixing of vesicles with PEG. The ability to measure fluorescence lifetimes on this time scale has allowed us to show that the of rate of lipid transfer increased dramatically at 35% PEG in both fusing and nonfusing vesicle systems. These results are interpreted in terms of a distinct interbilayer structure associated with intimate bilayer contact induced by high and potentially fusogenic concentrations of PEG.     

On the potential limitations of conventional sound metrics in quantifying perception of nonlinearly propagated noise

The use of conventional metrics to quantify the perception of nonlinearly propagated noise has been studied. Gaussian noise waveforms have been numerically propagated both linearly and nonlinearly, and from the resulting waveforms, several metrics are calculated. These metrics are overall, A-, C-, and D-weighted sound pressure levels, perceived noise level, Stevens Mark VII perceived loudness, Zwicker loudness, and sharpness. Informal listening demonstrations indicate that perceived differences in annoyance between linearly and nonlinearly propagated waveforms are substantial. Because the metrics studied seem inadequate in representing the perceived differences, rigorous subjective testing is encouraged to properly quantify and understand these differences.     

Everywhere invariant spaces of metrics and isometries

We define an everywhere invariant space of metricsU to be one for which the set of diffeomorphisms, which leaveU invariant, contains all the isometries of the individual metrics inU. We also generalize Killing's equation to a new equation, the invariance equation, which has as solutions those vector fields which leaveU invariant. By combining these ideas we give a new method for finding the isometries of a given metric.     

Tandem aza-Claisen rearrangement and ring-closing metathesis reactions: the stereoselective synthesis of functionalised carbocyclic amides

A one-pot, tandem process has been developed for the efficient synthesis of functionalised carbocyclic amides. A substituted cyclopentenyl trichloroacetamide was synthesised using a tandem thermal aza-Claisen rearrangement and RCM process, while an analogous cyclohexenyl trichloroacetamide was generated with high diastereoselectivity using a tandem MOM-ether directed metal-catalysed aza-Claisen rearrangement and RCM process.     

Fluorescence correlation spectroscopy using quantum dots: advances, challenges and opportunities

Semiconductor nanocrystals (quantum dots) have been increasingly employed in measuring the dynamic behavior of biomacromolecules using fluorescence correlation spectroscopy. This poses a challenge, because quantum dots display their own dynamic behavior in the form of intermittent photoluminescence, also known as blinking. In this review, the manifestation of blinking in correlation spectroscopy will be explored, preceded by an examination of quantum dot blinking in general.     

Dyeing uric acid crystals with methylene blue

The growth of anhydrous uric acid (UA) and uric acid dihydrate (UAD) crystals from supersaturated aqueous solutions containing methylene blue, a cationic organic dye, has been investigated. Low concentrations of dye molecules were found to be included in both types of crystal matrixes during the growth process. Incorporation of dye into UA crystals occurs with high specificity, affecting primarily [001] and [201] growth sectors, while UAD crystals grown from solutions of similar dye concentration show inclusion but little specificity. The orientation of the UA-trapped species was determined from polarization data obtained from visible light microspectrometry. To achieve charge neutrality, a second anionic species must also be included with the methylene blue into UA and UAD crystal matrices. Under high pH conditions, crystallization of 1:1 stoichiometric mixtures of methylene blue and urate yields methylene blue hexahydrate (MBU.6(H2O). The crystal structure of MBU.6(H2O) reveals continuous pi-pi stacks of planes of dye cations and urate anions mediated by water molecules. This structure provides an optimal geometry for methylene blue-urate pairs and additional support for the incorporation of these dimers in uric acid single-crystal matrices. The strikingly different inclusion patterns in UA and UAD demonstrate that subtle changes in the crystal surfaces and/or growth dynamics can greatly affect recognition events.     

An in situ atomic force microscopy study of uric acid crystal growth

Kidney stones are heterogeneous polycrystalline aggregates that can consist of several different building blocks. A significant number of human stones contain uric acid crystals as a crystalline component, though the molecular-level growth of this important biomaterial has not been previously well-characterized. In the present study, in situ atomic force microscopy (AFM) is used to investigate the real-time growth on the (100) surface of uric acid (UA) single crystals as a function of fundamental solution parameters. Layer-by-layer growth on UA (100) was found to be initiated at screw dislocation sites and to proceed via highly anisotropic rates which depend on the crystallographic direction. The smallest b-steps exhibited minimum heights corresponding to two molecular layers, while fast-moving c-steps more commonly showed monolayer heights. Growth kinetics measured under a range of flow rates, supersaturation levels, and pH values reveal linear trends in the growth kinetics, with faster growth attained in solutions with higher supersaturation and/or pH. The calculated kinetic parameters for UA growth derived from these experiments are in good agreement with the values reported for other crystal systems.     

Unfolding the torus: Oscillator geometry from time delays

Summary We present a simple method of plotting the trajectories of systems of weakly coupled oscillators. Our algorithm uses the time delays between the “firings” of the oscillators. For any system ofn weakly coupled oscillators there is an attracting invariantn-dimensional torus, and the attractor is a subset of this invariant torus. The invariant torus intersects a suitable codimension-1 surface of section at an (n−1)-dimensional torus. The dynamics ofn coupled oscillators can thus be reduced,in principle, to the study of Poincaré maps of the (n−1)-dimensional torus. This paper gives apractical algorithm for measuring then−1 angles on the torus. Since visualization of 3 (or higher) dimensional data is difficult we concentrate onn=3 oscillators. For three oscillators, a standard projection of the Poincaré map onto the plane yields a projection of the 2-torus which is 4-to-1 over most of the torus, making it difficult to observe the structure of the attractor. Our algorithm allows a direct measurement of the 2 angles on the torus, so we can plot a 1-to-1 map from the invariant torus to the “unfolded torus” where opposite edges of a square are identified. In the cases where the attractor is a torus knot, the knot type of the attractor is obvious in our projection.