NMR studies of caldesmon-calmodulin interactions

The binding of the calcium-regulatory protein calmodulin (CaM) to caldesmon (CaD) contributes to the regulation of smooth muscle contraction. Two regions of caldesmon have been identified as putative calmodulin-binding domains. We have earlier reported on the binding of one of these domains to calmodulin (Zhang & Vogel (1994) Biochemistry 33, 1163-1171). Here we have studied the binding of CaM to synthetic peptides of CaD which contain: (1) both the first and second CaM-binding domains; (2) the second CaM-binding domain; and (3) the sequence between the first and second CaM-binding domains. Two-dimensional transferred nuclear Overhauser enhancement proton NMR measurements as well as circular dichroism studies of a 22-residue peptide NKETAGLKVGVSSRINEWLTK, which contains the second CaM-binding domain, show that only the C-terminal half of the peptide becomes alpha-helical upon binding to CaM. Somewhat surprisingly, the shorter 9-residue peptide SRINEWLTK was sufficient to form a 1:1 complex with CaM; this peptide appears to bind as a 3(10)-helix. Proton-carbon-13 correlation NMR titration studies with specifically labeled [methyl-13C]methionine CaM were used to study the participation of the hydrophobic regions in both domains of the dumbbell shaped CaM in peptide binding. Binding of a 54-residue CaD peptide containing both CaM-binding domains affects all the 8 Met residues in the two hydrophobic domains of CaM (only Met 76 in the linker region of CaM is not involved), while binding of the second CaM-binding domain of CaD influences principally Met 51, 71, and Met 124, 144. Simultaneous binding to CaM of two peptides comprising the first and the second CaM-binding domains also caused changes to all Met residues except Met 76. Taken together, these data demonstrate that both CaM-binding domains of CaD can bind simultaneously to the two hydrophobic regions of CaM.     

Some potential applications for intercalation compounds of graphite with high electrical conductivity

Intercalation compounds of graphite of the acceptor type have potential engineering applications because of their attractive electrical conductivity properties. Two kinds of applications are considered in this paper. The first concerns a composite, formed by enclosing an intercalation compound synthesized from high quality crystalline graphite in a matrix of copper. With this form of composite it is found that there are both intrinsic and extrinsic advantages pertaining to the use of a material that has a conductivity higher than and a density lower than that of copper. The second form is a composite compound of intercalated graphite fibers contained in a matrix of epoxy. Extraordinary advantages in this case result from the fact that while intercalation of the fibers produces an order of magnitude increase in their electrical conductivity, when these fibers are incorporated into an epoxy matrix, the composite conductivity is increased by two orders of magnitude over its pristine fiber counterpart. It is projected that these desirable electrical conductivity characteristics portend large scale uses for the acceptor compounds of graphite as substitutes for the present standard conductors, and as a way of upgrading the performance of carbon/graphite materials.     

Sequential switch of biomineral crystal morphology using trivalent ions

Many biominerals are laminated such that crystal shape or habit changes from layer to layer thus yielding exquisitely designed composite materials with tightly controlled properties. Although lamination in biominerals is usually performed using peptides and proteins, here we introduce a new strategy by which sequential addition or depletion of inorganic trivalent ions in a supersaturated solution can be used to switch the surface morphology of calcium oxalate monohydrate (COM) back and forth, resulting in either the growth of flat crystalline sheets or of nanostructures oriented perpendicular to the surface. We propose that the occupation of a Ca(2+) site by Eu(3+) ion switches the orientation of the COM unit cell. The need to compensate the third charge forces coordination of Eu(3+) to an additional oxalate ion ((-)OOC-COO(-)) in an orientation that is not compatible with the initial unit cell. This mechanism of switching the orientation of the unit cell is unique, as it does not involve the use of expensive and thermally labile biomolecules. Suggestions of how to extend this strategy to engineer non-biological nanocomposites are given.     

Traffic sense--which factors influence the skill to predict the development of traffic scenes?

A study was conducted to evaluate the skill to predict the development of traffic situations. A stop-controlled intersection was filmed over several days, and 12 scenes with varying traffic complexity were selected. In half of the scenes, the traffic rules were violated, in half of the scenes, the rules were observed. A total of 36 participants were asked to watch the scenes and predict how the scene would most likely develop in the 2s after the film was paused. Additionally, the participants rated how certain they were about their prediction, and how complex and dangerous they assessed the scenes to be. With the method used here, experienced drivers were not found to make more correct predictions of situational development, and no difference in skill to predict could be found between genders. Nevertheless, more experienced drivers were more certain in their judgements and evaluated the situations on average as less complex and dangerous than did less experienced drivers. Scenes in which the traffic rules were violated were more difficult to predict correctly. The scenes in which the participants predicted violations were rated as more complex and dangerous. It is concluded that the low-cost method used here is more useful for examining which scenes are generally easy or difficult to predict and how they are experienced subjectively than to investigate differences in performance for different driver categories.     

A comparison of headway and time to collision as safety indicators

The two safety indicators "headway" and "time to collision (TTC)" are discussed and compared with respect to their usefulness in determining the safety of different traffic situations, like different locations in a junction. Over a 6-day-period traffic flow measures were taken in a four-way junction with stop signs on the minor road. It was found that for vehicles in a car following situation headway and TTC are independent of each other. The percentage of small headways is relatively constant across different locations in the junction, while the percentage of small TTC values varies between different locations. It is recommended to use headway for enforcement purposes, because small headways generate potentially dangerous situations. TTC, on the other hand, should be used when a certain traffic environment is to be evaluated in terms of safety, because it indicates the actual occurrences of dangerous situations.     

Micropositioning and microscopic observation of individual picoliter-sized containers within SU-8 microchannels

We describe the fabrication and application of a bioanalytical chip, made of SU-8 photoresist, comprising integrated, high aspect-ratio microfluidic channels, suitable to manipulate and investigate vesicles, cell fragments and biological cells. A central micrometer-sized aperture allows electrical particle counting and planar membrane experiments, microfluids allow (sub)micrometer-sized objects to be transported and addressed with different chemicals. Here we show how lipid vesicles are positioned with micrometer precision within the micro-channels by means of pressure and electrophoretic movement. Our approach is suited for controlling and investigating (bio)chemical synthesis and cellular signalling processes in ultrasmall individual vesicles by electro-optical techniques.     

Scintillation light produced by low-energy beams of highly-charged ions

Measurements have been performed of scintillation light intensities emitted from various inorganic scintillators irradiated with low-energy beams of highly-charged ions from an electron beam ion source (EBIS) and an electron cyclotron resonance ion source (ECRIS). Beams of xenon ions Xe^q+ with various charge states between q = 2 and q = 18 have been used at energies between 5 and 17.5 keV per charge generated by the ECRIS. The intensity of the beam was typically varied between 1 and 100 nA. Beams of highly charged residual gas ions have been produced by the EBIS at 4.5 keV per charge and with low intensities down to 100 pA. The scintillator materials used are flat screens of P46 YAG and P43 phosphor. In all cases, scintillation light emitted from the screen surface was detected by a CCD camera. The scintillation light intensity has been found to depend linearly on the kinetic ion energy per time deposited into the scintillator, while up to q = 18 no significant contribution from the ions’ potential energy was found. We discuss the results on the background of a possible use as beam diagnostics, e.g. for the new HITRAP facility at GSI, Germany.     

Male gender role conflict and willingness to seek counseling: Testing a mediation model on college-aged men.

Men who experience negative consequences of their socialized gender roles--that is, have greater gender role conflict--report less positive attitudes and willingness to seeking counseling. Using structural equation modeling with data from 575 undergraduate men, the authors examined 3 mediators (self-stigma associated with seeking counseling, tendency to disclose distressing information, and attitudes toward seeking counseling) regarding the link between gender role conflict and willingness to seek counseling for psychological and interpersonal concerns. Results indicated that this link was partially mediated by these 3 factors. Men experiencing greater gender role conflict were more likely to self-stigmatize and less likely to self-disclose. High self-stigma and less disclosure then led to less positive attitudes and subsequently to less willingness to seek counseling. (PsycINFO Database Record (c) 2010 APA, all rights reserved)