Shear stress,vascular remodeling and neointimal formation

The role of shear stress in atherosclerosis has been well documented. However, its role in restenosis was underexposed. In this paper a novel in vivo measuring technique and several of its applications related to restenosis will be described. The technique consists of a combination of 3D reconstruction of blood vessels and computational fluid dynamics (CFD). The 3D imaging techniques use either of 3D intravascular ultrasound (IVUS) as a stand-alone technique or a fusion of biplane angiography and IVUS (ANGUS). CFD is applied in order to relate local shear stress distribution to the morphology of the vessel wall. In the applications of these techniques it will be demonstrated that shear stress plays a role in the prediction of neointimal formation in in-stent restenosis and in vascular remodeling after balloon angioplasty. Attempts to locally increase shear stress by a newly developed flow divider indicate that shear stress reduce in-stent neointimal formation by 50%.     

Regional differences in hyoid muscle activity and length dynamics during mammalian head shaking

The sternohyoid (SH) and geniohyoid (GH) are antagonist strap muscles that are active during a number of different behaviors, including sucking, intraoral transport, swallowing, breathing, and extension/flexion of the neck. Because these muscles have served different functions through the evolutionary history of vertebrates, it is quite likely they will have complex patterns of electrical activity and muscle fiber contraction. Different regions of the SH exhibit different contraction and activity patterns during a swallow. We examined the dynamics of the SH and GH muscles during an unrestrained, and vigorous head shaking behavior in an animal model of human head, neck, and hyolingual movement. A gentle touch to infant pig ears elicited a head shake of several revolutions. Using sonomicrometry and intramuscular EMG, we measured regional (within) muscle strain and activity in SH and GH. We found that EMG was consistent across three regions (anterior, belly, and posterior) of each muscle. Changes in muscle length, however, were more complex. In the SH, mid-belly length-change occurred out-of-phase with the anterior and posterior end regions, but with a zero lag timing; the anterior region shortened before the posterior. In the GH, the anterior region shortened before and out-of-phase with the mid-belly and posterior regions. Head shaking is a relatively simple reflex behavior, yet the underlying patterns of muscle length dynamics and EMG activity are not. The regional complexity in SH and GH, similar to regionalization of SH during swallowing, suggests that these anatomically simple hyoid strap muscles have more complex function than textbooks often suggest.     

Strain distribution over plaques in human coronary arteries relates to shear stress

Once plaques intrude into the lumen, the shear stress they are exposed to alters with hitherto unknown consequences for plaque composition. We investigated the relationship between shear stress and strain, a marker for plaque composition, in human coronary arteries. We imaged 31 plaques in coronary arteries with angiography and intravascular ultrasound. Computational fluid dynamics was used to obtain shear stress. Palpography was applied to measure strain. Each plaque was divided into four regions: upstream, throat, shoulder, and downstream. Average shear stress and strain were determined in each region. Shear stress in the upstream, shoulder, throat, and downstream region was 2.55+/-0.89, 2.07+/-0.98, 2.32+/-1.11, and 0.67+/-0.35 Pa, respectively. Shear stress in the downstream region was significantly lower. Strain in the downstream region was also significantly lower than the values in the other regions (0.23+/-0.08% vs. 0.48+/-0.15%, 0.43+/-0.17%, and 0.47+/-0.12%, for the upstream, shoulder, and throat regions, respectively). Pooling all regions, dividing shear stress per plaque into tertiles, and computing average strain showed a positive correlation; for low, medium, and high shear stress, strain was 0.23+/-0.10%, 0.40+/-0.15%, and 0.60+/-0.18%, respectively. Low strain colocalizes with low shear stress downstream of plaques. Higher strain can be found in all other plaque regions, with the highest strain found in regions exposed to the highest shear stresses. This indicates that high shear stress might destabilize plaques, which could lead to plaque rupture.     

Display of Passenger Proteins on the Surface of Escherichia coli K-12 by the Enterohemorrhagic E. coli Intimin EaeA

Intimins are members of a family of bacterial adhesins from pathogenic Escherichia coli which specifically interact with diverse eukaryotic cell surface receptors. The EaeA intimin from enterohemorrhagic E. coli O157:H7 contains an N-terminal transporter domain, which resides in the bacterial outer membrane and promotes the translocation of four C-terminally attached passenger domains across the bacterial cell envelope. We investigated whether truncated EaeA intimin lacking two carboxy-terminal domains could be used as a translocator for heterologous passenger proteins. We found that a variant of the trypsin inhibitor Ecballium elaterium trypsin inhibitor II (EETI-II), interleukin 4, and the Bence-Jones protein REI(v) were displayed on the surface of E. coli K-12 via fusion to truncated intimin. Fusion protein net accumulation in the outer membrane could be regulated over a broad range by varying the cellular amount of suppressor tRNA that is necessary for translational readthrough at an amber codon residing within the truncated eaeA gene. Intimin-mediated adhesion of the bacterial cells to eukaryotic target cells could be mimicked by surface display of a short fibrinogen receptor binding peptide containing an arginine-glycine-aspartic acid sequence motif, which promoted binding of E. coli K-12 to human platelets. Cells displaying a particular epitope sequence fused to truncated intimin could be enriched 200,000-fold by immunofluorescence staining and fluorescence-activated cell sorting in three sorting rounds. These results demonstrate that truncated intimin can be used as an anchor protein that mediates the translocation of various passenger proteins through the cytoplasmic and outer membranes of E. coli and their exposure on the cell surface. Intimin display may prove a useful tool for future protein translocation studies with interesting biological and biotechnological ramifications.     

Adaptive significance of synchronous chorusing in an acoustically signalling wolf spider

Synchronous sexual signalling is a behavioural phenomenon that has received considerable theoretical interest, but surprisingly few empirical tests have been conducted. Here, we present a set of experiments designed to determine (i) whether the sexual signalling of the drumming wolf spider Hygrolycosa rubrofasciata is synchronous, and (ii) whether the synchrony may have evolved through female preference. Using controlled playback experiments, we found that males actively synchronized their drumming bouts with other males and females significantly preferred closely synchronized drumming clusters compared with loose clusters. In loose clusters, the first drumming signals attracted the most female responses, whereas in close clusters, the last drumming signals were the most heeded. We suggest that this female preference for the last drummer can maintain male synchronous signalling in H. rubrofasciata.     

Novel neuronal and astrocytic mechanisms in thalamocortical loop dynamics

In this review, we summarize three sets of findings that have recently been observed in thalamic astrocytes and neurons, and discuss their significance for thalamocortical loop dynamics. (i) A physiologically relevant 'window' component of the low-voltage-activated, T-type Ca(2+) current (I(Twindow)) plays an essential part in the slow (less than 1 Hz) sleep oscillation in adult thalamocortical (TC) neurons, indicating that the expression of this fundamental sleep rhythm in these neurons is not a simple reflection of cortical network activity. It is also likely that I(Twindow) underlies one of the cellular mechanisms enabling TC neurons to produce burst firing in response to novel sensory stimuli. (ii) Both electrophysiological and dye-injection experiments support the existence of gap junction-mediated coupling among young and adult TC neurons. This finding indicates that electrical coupling-mediated synchronization might be implicated in the high and low frequency oscillatory activities expressed by this type of thalamic neuron. (iii) Spontaneous intracellular Ca(2+) ([Ca(2+)](i)) waves propagating among thalamic astrocytes are able to elicit large and long-lasting N-methyl-D-aspartate-mediated currents in TC neurons. The peculiar developmental profile within the first two postnatal weeks of these astrocytic [Ca(2+)](i) transients and the selective activation of these glutamate receptors point to a role for this astrocyte-to-neuron signalling mechanism in the topographic wiring of the thalamocortical loop. As some of these novel cellular and intracellular properties are not restricted to thalamic astrocytes and neurons, their significance may well apply to (patho)physiological functions of glial and neuronal elements in other brain areas.     

Coronary stent implantation changes 3-D vessel geometry and 3-D shear stress distribution

Mechanisms of in-stent restenosis are not fully understood. Shear stress is known to play a role in plaque and thrombus formation and is sensitive to changes in regional vessel geometry. Hence, we evaluated the regional changes in 3-D geometry and shear stress induced by stent placement in coronary arteries of pigs.Methods. 3-D reconstruction was performed, applying a combined angiographic and IVUS technique (ANGUS), from seven Wallstents (diameter 3.5 (n=3) and 5mm (n=4)), which were implanted in seven coronary arteries of five pigs. This 3-D geometry was used to calculate locally the curvature, while the shear stress distribution was obtained by computational fluid dynamics. Local changes in shear stress were obtained at the entrance and exit of the stent for baseline (0. 65+/-0.22 ml/s) and hyperemic flow (2.60+/-0.86 ml/s) conditions. Results. After stent implantation, the curvature increased by 121% at the entrance and by 100% at the exit of the stent, resulting in local changes in shear stress. In general, at the entrance of the stent local maxima in shear stress were generated, while at the exit both local maxima and minima in shear stress were observed (p<0.05). Additionally, the shear stress at the entrance and exit of the stent were correlated with the local curvature (r: 0.30-0.84).Conclusion. Stent implantation changes 3-D vessel geometry in such a way that regions with decreased and increased shear stress occur close to the stent edges. These changes might be related to the asymmetric patterns of in-stent restenosis.     

Influence of catheter design on lumen wall temperature distribution in intracoronary thermography

Intracoronary thermography is a currently used vulnerable plaque detection method. We studied how catheter design and catheter location influence the temperature readings, and thus its capacity to detect vulnerable plaques. Finite element calculations were performed on geometries representing the coronary artery, the vulnerable plaque and the catheter. Catheter material, diameter and location with respect to the plaque were varied. Both flow and no-flow situations were studied. Maximal lumen wall temperature difference without a catheter (DeltaT=0.12 degrees C, flow=75 cm(3) min(-1)) was considered the reference. Presence of a 1.0mm nitinol catheter right under the plaque increased DeltaT to 0.14 degrees C, whereas a 1.0 mm polyurethane catheter increased DeltaT to 0.51 degrees C. The location at which a thermosensitive element should be placed for most optimal temperature readings during a pullback was shown to lie at the catheter edge for the nitinol catheter and at 1.1mm from the catheter edge for the polyurethane catheter. Temperature readings decreased to background temperature when the catheter was in close proximity but not overlapping the plaque. DeltaT decreased approximately by 70% when a gap of 0.2 mm existed between the catheter and the lumen wall. Occlusion of blood flow increased DeltaT values in all cases, but most pronounced for nitinol catheters. A polyurethane catheter increased the temperature readings, since its heat conductivity is lower than that of blood, which makes it a very good choice for heat source detection. Catheter design can contribute to enhanced temperature readings and thus can enable more optimal vulnerable plaque detection.