Imaging cytoplasmic cAMP in mouse brainstem neurons

Background  

cAMP is an ubiquitous second messenger mediating various neuronal functions, often as a consequence of increased intracellular Ca²⁺ levels. While imaging of calcium is commonly used in neuroscience applications, probing for cAMP levels has not yet been performed in living vertebrate neuronal tissue before.     

A simple model for a two-stage chemical reaction with diffusion

We analyse the evolution of a two-stage chemical reaction betweentwo neighbouring plumes of reactants. Under the assumption thatthe plumes are approximately Gaussian we derive a system ofordinary differential equations for the total amount, the centroidand the variance of each reactant. We compare the solution ofthese equations with full numerical simulation of the reaction.Excellent agreement is obtained, with solution of the near-Gaussianmodel requiring considerably less computational effort thanthe full simulations. Of key importance is the yield of thereaction, and we discuss this feature in particular.     

Instantaneous fluctuation velocity and skewness distributions upstream of transition onset

The development of streamwise orientated disturbances through the boundary layer thickness prior to transition onset for zero-pressure gradient boundary layer flow under the influence %Tu = 4.2 is presented. The analysis concentrates on the development of the maximum positive and negative of the fluctuation velocity in order to gain further insight into the transition process. The average location of the peak negative fluctuation velocity over a range of Reynolds numbers was measured in the upper portion of the boundary layer at y/δ ≈ 0.6, whereas the location of the peak positive value was measured at y/δ ≈ 0.3. The disturbance magnitude of the negative fluctuation velocity increased beyond that of the positive as transition onset approached. The distribution and disturbance magnitude of the maximum positive and negative fluctuation velocities indicate that the initiation of transition may occur on the low-speed components of the flow that are lifted up to the upper region of the boundary layer. This is in qualitative agreement with recent direct numerical simulations on the breakdown of the flow on the lifted low-speed streaks near the boundary layer edge. The results presented in this investigation also demonstrate the increased physical insight gained by examining the distributions of the maximum positive and negative of the streamwise fluctuation velocity component associated with the low- and high-speed streaks, compared to time-averaged values, in determining what structures cause the breakdown to turbulence.     

Determining a 'safe' high-mass limit in matrix-assisted laser desorption/ionisation time-of-flight mass spectra of coal derived materials with reference to instrument noise

Three methods for determining a 'safe' estimate for high-mass limits of MALDI spectra of coal derived liquids were explored, using a sample of coal-tar pitch and its pyridine-insoluble fraction. Co-addition of increasing numbers of single-shot spectra (10, 30, 50 and 100 pulses) showed visually observable reductions in noise levels, consistent with robust and statistically meaningful signals. Three separate types of post-acquisition calculation were used to identify high-mass limits of the spectra. (i) A literature method indicated high-mass limits similar to those observed visually-as a shift from baseline at the highest masses, nearly 350 000 u for the coal tar pitch and about 390 000 u for its pyridine insoluble fraction. (ii) Comparing instrument signal with pre-selected multiples of the standard deviation, upper mass estimates of between 40-60 000 u for the coal-tar pitch and about 95 000 u for its pyridine-insoluble fraction were found. (iii) Calculation of the slope was used to identify 'lift-off' of the spectrum from baseline. The angle between the smoothed spectrum and the baseline was matched to a pre-selected value (e.g. 0.5 degrees and 1 degrees ). However, the arbitrary specification of the key parameter did not establish this last method on a firm basis. The choice of a criterion for estimating high-mass limits of MALDI spectra remains a semi-quantitative procedure; a reasonably conservative high-mass limit may be estimated by comparison of signal with five times the standard deviation. However, evaluation of size exclusion chromatograms of the present samples using polystyrene standards suggests that molecular mass distributions of pitch samples arrived at by MALDI mass spectrometry are, at least partly, determined by the limitations of available instruments. Copyright 1999 John Wiley & Sons, Ltd.