Modeling intracranial aneurysm stability and growth: an integrative mechanobiological framework for clinical cases

We present a novel patient-specific fluid-solid-growth framework to model the mechanobiological state of clinically detected intracranial aneurysms (IAs) and their evolution. The artery and IA sac are modeled as thick-walled, non-linear elastic fiber-reinforced composites. We represent the undulation distribution of collagen fibers: the adventitia of the healthy artery is modeled as a protective sheath whereas the aneurysm sac is modeled to bear load within physiological range of pressures. Initially, we assume the detected IA is stable and then consider two flow-related mechanisms to drive enlargement: (1) low wall shear stress; (2) dysfunctional endothelium which is associated with regions of high oscillatory flow. Localized collagen degradation and remodelling gives rise to formation of secondary blebs on the aneurysm dome. Restabilization of blebs is achieved by remodelling of the homeostatic collagen fiber stretch distribution. This integrative mechanobiological modelling workflow provides a step towards a personalized risk-assessment and treatment of clinically detected IAs.

     

Layer-dependent role of collagen recruitment during loading of the rat bladder wall

In this work, we re-evaluated long-standing conjectures as to the source of the exceptionally large compliance of the bladder wall. Whereas these conjectures were based on indirect measures of loading mechanisms, in this work we take advantage of advances in bioimaging to directly assess collagen fibers and wall architecture during biaxial loading. A custom biaxial mechanical testing system compatible with multiphoton microscopy was used to directly measure the layer-dependent collagen fiber recruitment in bladder tissue from 9 male Fischer rats (4 adult and 5 aged). As for other soft tissues, the bladder loading curve was exponential in shape and could be divided into toe, transition and high stress regimes. The relationship between collagen recruitment and loading curves was evaluated in the context of the inner (lamina propria) and outer (detrusor smooth muscle) layers. The large extensibility of the bladder was found to be possible due to folds in the wall (rugae) that provide a mechanism for low resistance flattening without any discernible recruitment of collagen fibers throughout the toe regime. For more extensible bladders, as the loading extended into the transition regime, a gradual coordinated recruitment of collagen fibers between the lamina propria layer and detrusor smooth muscle layer was found. A second important finding was that wall extensibility could be lost by premature recruitment of collagen in the outer wall that cut short the toe region. This change was correlated with age. This work provides, for the first time, a mechanistic understanding of the role of collagen recruitment in determining bladder extensibility and capacitance.     

The dynamics of protein kinase B regulation during B cell antigen receptor engagement.

This study has used biochemistry and real time confocal imaging of green fluorescent protein (GFP)-tagged molecules in live cells to explore the dynamics of protein kinase B (PKB) regulation during B lymphocyte activation. The data show that triggering of the B cell antigen receptor (BCR) induces a transient membrane localization of PKB but a sustained activation of the enzyme; active PKB is found in the cytosol and nuclei of activated B cells. Hence, PKB has three potential sites of action in B lymphocytes; transiently after BCR triggering PKB can phosphorylate plasma membrane localized targets, whereas during the sustained B cell response to antigen, PKB acts in the nucleus and the cytosol. Membrane translocation of PKB and subsequent PKB activation are dependent on BCR activation of phosphatidylinositol 3-kinase (PI3K). Moreover, PI3K signals are both necessary and sufficient for sustained activation of PKB in B lymphocytes. However, under conditions of continuous PI3K activation or BCR triggering there is only transient recruitment of PKB to the plasma membrane, indicating that there must be a molecular mechanism to dissociate PKB from sites of PI3K activity in B cells. The inhibitory Fc receptor, the FcgammaRIIB, mediates vital homeostatic control of B cell function by recruiting an inositol 5 phosphatase SHIP into the BCR complex. Herein we show that coligation of the BCR with the inhibitory FcgammaRIIB prevents membrane targeting of PKB. The FcgammaRIIB can thus antagonize BCR signals for PKB localization and prevent BCR stimulation of PKB activity which demonstrates the mechanism for the inhibitory action of the FcgammaRIIB on the BCR/PKB response.     

Evolving mechanical properties of a model of abdominal aortic aneurysm

The novel three-dimensional (3D) mathematical model for the development of abdominal aortic aneurysm (AAA) of Watton et al. Biomech Model Mechanobiol 3(2): 98–113, (2004) describes how changes in the micro-structure of the arterial wall lead to the development of AAA, during which collagen remodels to compensate for loss of elastin. In this paper, we examine the influence of several of the model’s material and remodelling parameters on growth rates of the AAA and compare with clinical data. Furthermore, we calculate the dynamic properties of the AAA at different stages in its development and examine the evolution of clinically measurable mechanical properties. The model predicts that the maximum diameter of the aneurysm increases exponentially and that the ratio of systolic to diastolic diameter decreases from 1.13 to 1.02 as the aneurysm develops; these predictions are consistent with physiological observations of Vardulaki et al. Br J Surg 85:1674–1680 (1998) and Lanne et al. Eur J Vasc Surg 6:178–184 (1992), respectively. We conclude that mathematical models of aneurysm growth have the potential to be useful, noninvasive diagnostic tools and thus merit further development.     

Novel sites in the p65 subunit of NF-kappaB interact with TFIIB to facilitate NF-kappaB induced transcription

The citM gene from Lactococcus lactis CRL264 was demonstrated to encode for an oxaloacetate decarboxylase. The enzyme exhibits high levels of similarity to malic enzymes (MEs) from other organisms. CitM was expressed in Escherichia coli, purified and its oxaloacetate decarboxylase activity was demonstrated by biochemical and genetic studies. The highest oxaloacetate decarboxylation activity was found at low pH in the presence of manganese, and the K_m value for oxaloacetate was 0.52 ± 0.03 mM. However, no malic activity was found for this enzyme. Our studies clearly show a new group of oxaloacetate decarboxylases associated with the citrate fermentation pathway in gram-positive bacteria. Furthermore, the essential catalytic residues were found to be conserved in all members of the ME family, suggesting a common mechanism for oxaloacetate decarboxylation.     

Simulation of coronary artery revascularization

Simulation of the commonly constructed geometries of aorto-coronary bypass anastomoses was carried out using especially fabricated distensible tubes and a pulsatile pump. The system pressure was maintained between 80 and 120 mmHg. The total mean flow was set at 250 ml min-1 (Reynolds number of 200) and the pulsatile frequency was varied from 0 to 2 Hz. A water-glycerine mixture having a density and viscosity similar to that of blood was used throughout. A 16 mm film of the front of black dye injected proximal to the anastomosis was made as the dye approached and passed through the anastomosis. Anastomotic geometries consisted of: end to side, parallel, 45 degree angle, and 90 degree angle. Stenoses, located in the tube representing the coronary artery, were simulated using a bevelled insert which represented an 80-85% area reduction. Flow visualization revealed that distensible tubes gave more realistic flow patterns than rigid tubes, a result particularly evident when a stenosis was present. Pulsatile flow demonstrated considerably more mixing than steady flow. The use of pulsatile flow in distensible tubing with a partial stenosis showed retrograde flow through the stenosis which was not evident for either steady flow or for flow in rigid tubing. The flow at the anastomatic site of the graft having an angle of 0 degrees showed a jetting action with a zone of recirculating fluid being present whereas for a 90 degree graft a distinct helical flow was formed distal to the anastomosis.     

The Spatial Distribution of the Exocyst and Actin Cortical Patches Is Sufficient To Organize Hyphal Tip Growth

In the hyphal tip of Candida albicans we have made detailed quantitative measurements of (i) exocyst components, (ii) Rho1, the regulatory subunit of (1,3)-β-glucan synthase, (iii) Rom2, the specialized guanine-nucleotide exchange factor (GEF) of Rho1, and (iv) actin cortical patches, the sites of endocytosis. We use the resulting data to construct and test a quantitative 3-dimensional model of fungal hyphal growth based on the proposition that vesicles fuse with the hyphal tip at a rate determined by the local density of exocyst components. Enzymes such as (1,3)-β-glucan synthase thus embedded in the plasma membrane continue to synthesize the cell wall until they are removed by endocytosis. The model successfully predicts the shape and dimensions of the hyphae, provided that endocytosis acts to remove cell wall-synthesizing enzymes at the subapical bands of actin patches. Moreover, a key prediction of the model is that the distribution of the synthase is substantially broader than the area occupied by the exocyst. This prediction is borne out by our quantitative measurements. Thus, although the model highlights detailed issues that require further investigation, in general terms the pattern of tip growth of fungal hyphae can be satisfactorily explained by a simple but quantitative model rooted within the known molecular processes of polarized growth. Moreover, the methodology can be readily adapted to model other forms of polarized growth, such as that which occurs in plant pollen tubes.     

Modelling evolution and the evolving mechanical environment of saccular cerebral aneurysms

A fluid–solid-growth (FSG) model of saccular cerebral aneurysm evolution is developed. It utilises a realistic two-layered structural model of the internal carotid artery and explicitly accounts for the degradation of the elastinous constituents and growth and remodelling (G&R) of the collagen fabric. Aneurysm inception is prescribed: a localised degradation of elastin results in a perturbation in the arterial geometry; the collagen fabric adapts, and the artery achieves a new homeostatic configuration. The perturbation to the geometry creates an altered haemodynamic environment. Subsequent degradation of elastin is explicitly linked to low wall shear stress (WSS) in a confined region of the arterial domain. A sidewall saccular aneurysm develops, the collagen fabric adapts and the aneurysm stabilises in size. A quasi-static analysis is performed to determine the geometry at diastolic pressure. This enables the cyclic stretching of the tissue to be quantified, and we propose a novel index to quantify the degree of biaxial stretching of the tissue. Whilst growth is linked to low WSS from a steady (systolic) flow analysis, a pulsatile flow analysis is performed to compare steady and pulsatile flow parameters during evolution. This model illustrates the evolving mechanical environment for an idealised saccular cerebral aneurysm developing on a cylindrical parent artery and provides the guidance to more sophisticated FSG models of aneurysm evolution which link G&R to the local mechanical stimuli of vascular cells.     

Vesicle sizing: Number distributions by dynamic light scattering

A procedure is described which optimizes nonnegative least squares and exponential sampling fitting methods for analysis of dynamic light scattering (DLS) data from aqueous suspensions of vesicle/liposome systems. This approach utilizes a Rayleigh-Gans-Debye form factor for a coated sphere and yields number distributions which can be compared directly to distributions obtained by freeze-fracture electron microscopy (EM). Excellent agreement between the DLS and EM results are obtained for vesicle size distributions in the 100-200-nm range.