A Multivalent Ligand for the Mannose‐6‐Phosphate Receptor for Endolysosomal Targeting of an Activity‐Based Probe

The ubiquitously expressed mannose‐6‐phosphate receptors (MPRs) are a promising class of receptors for targeted compound delivery into the endolysosomal compartments of a variety of cell types. The development of a synthetic, multivalent, mannose‐6‐phosphate (M6P) glycopeptide‐based MPR ligand is described. The conjugation of this ligand to fluorescent DCG‐04, an activity‐based probe for cysteine cathepsins, enabled fluorescent readout of its receptor‐targeting properties. The resulting M6P‐cluster–BODIPY–DCG‐04 probe was shown to efficiently label cathepsins in cell lysates as well as in live cells. Furthermore, the introduction of the 6‐O‐phosphates leads to a completely altered uptake profile in COS and dendritic cells compared to a mannose‐containing ligand. Competition with mannose‐6‐phosphate abolished all uptake of the probe in COS cells, and we conclude that the mannose‐6‐phosphate cluster targets the MPR and ensures the targeted delivery of cargo bound to the cluster into the endolysosomal pathway.     

Heritability of Stroop and flanker performance in 12-year old children

Background  

There is great interest in appropriate phenotypes that serve as indicator of genetically transmitted frontal (dys)function, such as ADHD. Here we investigate the ability to deal with response conflict, and we ask to what extent performance variation on response interference tasks is caused by genetic variation. We tested a large sample of 12-year old monozygotic and dizygotic twins on two well-known and closely related response interference tasks; the color Stroop task and the Eriksen flanker task. Using structural equation modelling we assessed the heritability of several performance indices derived from those tasks.     

Transients in flow and local heat transfer due to a pressure wave in pipe flow

The effect of a pressure wave on the turbulent flow and heat transfer in a rectangular air flow channel has been experimentally studied for fast transients, occurring due to a sudden increase of the main flow by an injection of air through the wall. A fast response measuring technique using a hot film sensor for the heat flux, a hot wire for the velocities and a pressure transducer have been developed. It was found that in the initial part of the transient the heat transfer change is independent of the Reynolds number. For the second part the change in heat transfer depends on thermal boundary layer thickness and thus on the Reynolds number. Results have been compared with a simple numerical turbulent flow and heat transfer model. The main effect on the flow could be well predicted. For the heat transfer a deviation in the initial part of the transient heat transfer has been found. From the turbulence measurements it has been found that a pressure wave does not influence the absolute value of the local turbulent velocity fluctuations. They could be considered to be frozen.Nomenclature A surface area (m²) - D diameter (m) - h heat transfer coefficient (Wm^–2 K^–1) - p pressure drop (Pa) - P pressure (Pa) - Q heat flow (W) - R tube radius (m) - T bulk temperature (K) - T _s surface temperature (K) - t time (s) - u velocity (m/s) - V voltage (V) - y distance from wall (m) - viscosity (N s m^–2) - kinematic viscosity (m^–2 s^–1) - density (kg m^–3) - _w wall shear stress (N m^–2) - Nu Nusselt number - Re Reynolds number     

Numerical computation of high Rayleigh number natural convection and prediction of hot radiator induced room air motion

The two-dimensional stationary turbulent buoyant flow and heat transfer in a cavity at high Rayleigh numbers was computed numerically. The k– turbulence model was used. The time-averaged equations for momentum, energy and continuity, which are coupled to the turbulence equations, were solved using a finite difference formulation. In order to validate the computer code, a comparison exercise was carried out. The test results are in good agreement with the internationally accepted benchmark solution. Grid-refinement shows the necessity of a very fine grid at high Rayleigh numbers with especially small grid-distances in the near-wall region. The computed boundary layer velocity profiles are in excellent agreement with available experimental data. The local heat transfer in the turbulent part of the boundary layers is predicted 20% too high. Computations were carried out for the natural convective flow in a room induced by a hot radiator and a cold window. Various radiator configurations and types of thermal boundary conditions were applied including thermal radiation interaction between surfaces.Nomenclature a thermal diffusivity (m²/s) - C constant in _t expression - D cavity dimensions (m) - g acceleration of gravity (m/s²) - G _k production/destruction of k by buoyancy (kg/ms³) - h enthalpy (J/kg) - IX index of grid point - k turbulent kinetic energy (m²/s²) - m dimensionless stratification parameter - Nu overall Nusselt number - Nu _y local Nusselt number - NX total number of grid points - p pressure (N/m²) - P _k production of k by shear stress (kg/ms³) - Q heat flux through wall (W/m) - Ra overall Rayleigh number - Ra _y local Rayleigh number - Re _t turbulent Reynolds number - S source term in -equation (kg/ms⁴) - S source term for - T _c, T _h temperatures of cold and hot walls (K) - T _s (y) stratification temperature on vertical mid-line (K) - T ₀ mean cavity temperature (K) - u, v horizontal and vertical velocity components (m/s) - u ₀ Brunt-Vaisälä velocity scale (m/s) - x, y horizontal and vertical coordinates (m) - non-linearity parameter for grid - coefficient of thermal expansion (l/K) - jet angle (°) - diffusivity for - S dissipation rate for turbulent kinetic energy (m²/s³) - variable to be solved - thermal conductivity (W/mK) - , _t kinematic and eddy viscosities (m²/s) - stream function (kg/ms) - density (kg/m³) - _k, , _t constants in k– model     

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.     

The contribution of high frequencies to human brain activity underlying horizontal localization of natural spatial sounds

Background  

In the field of auditory neuroscience, much research has focused on the neural processes underlying human sound localization. A recent magnetoencephalography (MEG) study investigated localization-related brain activity by measuring the N1m event-related response originating in the auditory cortex. It was found that the dynamic range of the right-hemispheric N1m response, defined as the mean difference in response magnitude between contralateral and ipsilateral stimulation, reflects cortical activity related to the discrimination of horizontal sound direction. Interestingly, the results also suggested that the presence of realistic spectral information within horizontally located spatial sounds resulted in a larger right-hemispheric N1m dynamic range. Spectral cues being predominant at high frequencies, the present study further investigated the issue by removing frequencies from the spatial stimuli with low-pass filtering. This resulted in a stepwise elimination of direction-specific spectral information. Interaural time and level differences were kept constant. The original, unfiltered stimuli were broadband noise signals presented from five frontal horizontal directions and binaurally recorded for eight human subjects with miniature microphones placed in each subject's ear canals. Stimuli were presented to the subjects during MEG registration and in a behavioral listening experiment.