Richard Manasseh

A Novel Mouse Model of Atherosclerotic Plaque Instability for Drug Testing and Mechanistic/Therapeutic Discoveries Using Gene and MicroRNA Expression Profiling

Rationale: The high morbidity/mortality of atherosclerosis is typically precipitated by plaque rupture and consequent thrombosis. However, research on underlying mechanisms and therapeutic approaches is limited by the lack of animal models that reproduce plaque instability observed in humans. Objective: Development and use of a mouse model of plaque rupture that reflects the end stage of human atherosclerosis. Methods and Results: On the basis of flow measurements and computational fluid dynamics, we applied a tandem stenosis to the carotid artery of apolipoprotein E–deficient mice on high-fat diet. At 7 weeks postoperatively, we observed intraplaque hemorrhage in ≈50% of mice, as well as disruption of fibrous caps, intraluminal thrombosis, neovascularization, and further characteristics typically seen in human unstable plaques. Administration of atorvastatin was associated with plaque stabilization and downregulation of monocyte chemoattractant protein-1 and ubiquitin. Microarray profiling of mRNA and microRNA (miR) and, in particular, its combined analysis demonstrated major differences in the hierarchical clustering of genes and miRs among nonatherosclerotic arteries, stable, and unstable plaques and allows the identification of distinct genes/miRs, potentially representing novel therapeutic targets for plaque stabilization. The feasibility of the described animal model as a discovery tool was established in a pilot approach, identifying a disintegrin and metalloprotease with thrombospondin motifs 4 (ADAMTS4) and miR-322 as potential pathogenic factors of plaque instability in mice and validated in human plaques. Conclusions: The newly described mouse model reflects human atherosclerotic plaque instability and represents a discovery tool toward the development and testing of therapeutic strategies aimed at preventing plaque rupture. Distinctly expressed genes and miRs can be linked to plaque instability.

Passive Acoustic Determination of Wave-Breaking Events and Their Severity across the Spectrum

Abstract A passive acoustic method of detecting breaking waves of different scales has been developed. The method also showed promise for measuring breaking severity. Sounds were measured by a subsurface hydrophone in various wind and wave states. A video record of the surface was made simultaneously. Individual sound pulses corresponding to the many individual bubble formations during wave-breaking events typically last only a few tens of milliseconds. Each time a sound-level threshold was exceeded, the acoustic signal was captured over a brief window typical of a bubble formation pulse, registering one count. Each pulse was also analyzed to determine the likely bubble size generating the pulse. Using the time series of counts and visual observations of the video record, the sound-level threshold that detected bubble formations at a rate optimally discriminating between breaking and nonbreaking waves was determined by a classification-accuracy analysis. This diagnosis of breaking waves was found to be ap...

Symmetric mode resonance of bubbles attached to a rigid boundary

Experimental results are compared with a theoretical analysis concerning wall effects on the symmetric mode resonance frequency of millimeter-sized air bubbles in water. An analytical model based on a linear coupled-oscillator approximation is used to describe the oscillations of the bubbles, while the method of images is used to model the effect of the wall. Three situations are considered: a single bubble, a group of two bubbles, and a group of three bubbles. The results show that bubbles attached to a rigid boundary have lower resonance frequencies compared to when they are in an infinite uniform liquid domain (referred to as free space). Both the experimental data and theoretical analysis show that the symmetric mode resonance frequency decreases with the number of bubbles but increases as the bubbles are moved apart. Discrepancies between theory and experiment can be explained by the fact that distortion effects due to buoyancy forces and surface tension were ignored. The data presented here are inte...

Production of monodispersed micron-sized bubbles at high rates in a microfluidic device

A polydimethylsiloxane microchip consisting of a T-junction microchannel network and a thin glass capillary has been developed for the generation of microbubbles. The glass capillary is used to produce an ultrathin gas jet and to controllably block the straight liquid channel, thereby increasing the local liquid velocity near the intersection. Liquid flow rate, liquid viscosity, gas pressure, and inner diameter of the gas jet are varied to investigate the effect on bubble generation. Bubbles with a diameter down to 4.5u2002μm can be produced at a high rate of 7.5 kHz using a capillary with an inner diameter of 2u2002μm.

Dynamics of pulsatile flow in fractal models of vascular branching networks.

Efficient regulation of blood flow is critically important to the normal function of many organs, especially the brain. To investigate the circulation of blood in complex, multi-branching vascular networks, a computer model consisting of a virtual fractal model of the vasculature and a mathematical model describing the transport of blood has been developed. Although limited by some constraints, in particular, the use of simplistic, uniformly distributed model for cerebral vasculature and the omission of anastomosis, the proposed computer model was found to provide insights into blood circulation in the cerebral vascular branching network plus the physiological and pathological factors which may affect its functionality. The numerical study conducted on a model of the middle cerebral artery region signified the important effects of vessel compliance, blood viscosity variation as a function of the blood hematocrit, and flow velocity profile on the distributions of flow and pressure in the vascular network.

Dynamics of dual-particles settling under gravity

Abstract As a first step towards understanding particle–particle interaction in fluid flows, the motion of two spherical particles settling in close proximity under gravity in Newtonian fluids was investigated experimentally for particle Reynolds numbers ranging from 0.01 to 2000. It was observed that particles repel each other for Re>0.1 and that the separation distance of settling particles is Reynolds number dependent. At lower Reynolds numbers, i.e. for Re The orientation preference of two spherical particles was found to be Reynolds number dependent. At higher Reynolds numbers, the line connecting the centres of the two particles is always horizontal, regardless of the way the two particles are launched. At lower Reynolds numbers, however, the particle centreline tends to tilt to an arbitrary angle, even of the two particles are launched in the horizontal plane. Because of the tilt, a side migration of the two particles was found to exist. A linear theory was developed to estimate the side migration velocity. It was found that the maximum side migration velocity is approximately 6% of the vertical settling velocity, in good agreement with the experimental results. Counter-rotating spinning of the two particles was observed and measured in the range of Re=0–10. Using the linear model, it is possible to estimate the influence of the tilt angle on the rate of rotation at low Reynolds numbers. Dual particles settle faster than a single particle at small Reynolds numbers but not at higher Reynolds numbers, because of particle separation. The variation of particle settling velocity with Reynolds number is presented. An equation which can be used to estimate the influence of tilt angle on particle settling velocity at low Reynolds number is also derived.

Chaotic micromixing in open wells using audio-frequency acoustic microstreaming

Mixing fluids for biochemical assays is problematic when volumes are very small (on the order of the 10 microL typical of single drops), which has inspired the development of many micromixing devices. In this paper, we show that micromixing is possible in the simple open wells of standard laboratory consumables using appropriate acoustic frequencies that can be applied using cheap, conventional audio components. Earlier work has shown that the phenomenon of acoustic microstreaming can mix fluids, provided that bubbles are introduced into a specially designed microchamber or that high-frequency surface acoustic wave devices are constructed. We demonstrate a key simplification: acoustic micromixing at audio frequencies by ensuring the system has a liquid-air interface with a small radius of curvature. The meniscus of a drop in a small well provided an appropriately small radius, and so an introduced bubble was not necessary. Microstreaming showed improvement over diffusion-based mixing by 1-2 orders of magnitude. Furthermore, significant improvements are attainable through the utilization of chaotic mixing principles, whereby alternating fluid flow patterns are created by applying, in sequence, two different acoustic frequencies to a drop of liquid in an open well.

Measurement of microbubble-induced acoustic microstreaming using microparticle image velocimetry

Micro particle image velocimetry (PIV) measurements of the velocity fields around oscillating gas bubbles in microfluidic geometries were undertaken. Two sets of experiments were performed. The first measured the acoustic microstreaming around a gas bubble with a radius of 195 μm attached to a wall in a chamber of 30 mm× 30 mm× 0.66 mm. Under acoustic excitation, vigorous streaming in the form of a circulation around on the bubble was observed. The streaming flow was highest near the surface of the bubble with velocities around 1mm/s measured. The velocity magnitude decreased rapidly with increasing distance from the bubble. The velocity field determined by micro-PIV matched the streaklines of the fluorescent particles very well. The second set of experiments measured the streaming at the interface between a trapped air bubble and water inside a microchannel of cross section 100 μm × 90 μm. The streaming flow was limited to within a short distance from the interface and was observed as a looping flow, moving towards the interface from the top and being circulated back from the bottom of the channel. The characteristic streaming velocity was in the order of 100 μm/s.

Acoustic microstreaming applied to batch micromixing

Experiments are presented in which acoustic microstreaming is investigated and applied to a batch micromixing case appropriate to a point-of-care pathology screening test. The flows presented can be created without complex engineering of contacts or surfaces in the microdevice, which could thus be made disposable. Fundamental flow patterns are measured with a micro-Particle-Image Velocimetry (micro-PIV) system, enabling a quantification of the fluiddynamical processes causing the flows. The design of micromixers based on this principle requires a quantification of the mixing. A simple technique based on digital image processing is presented that enables an assessment of the improvement in mixing due to acoustic microstreaming. The digital image processing technique developed was shown to be non-intrusive, convenient and able to generate useful quantitative data. Preliminary indications are that microstreaming can at least halve the time required to mix quantities of liquid typical of a point-of-care test, and significantly greater improvements seem feasible.

Modelling of embolus transport and embolic stroke

Cerebral microembolism may lead to the restriction of blood supply due to damaged blood vessel tissue (focal ischemia) which is increasingly seen as the cause of cognitive deterioration including Alzheimer’s disease and vascular dementia. The flow through fractal models of the peripheral vasculature of the Anterior Cerebral Arteries (ACA) and Middle Cerebral Arteries (MCA) was modelled. The multi-scale model of the cerebral vasculature was coupled with blood flow and embolus transport models. The model incorporated asymmetric bifurcation trees, embolus-vascular interactions and autoregulation. Simulations were carried out where the embolus deposition rate, embolus diameter and embolus introduction rate were varied. Increasing the embolus diameter and embolus introduction rate increased the number of blocked terminal arteries to a quasi steady-state. For a low embolus deposition rate the MCA and ACA territory had the same embolization dynamics, even though, the MCA was larger than the ACA. It was also found for a higher embolus deposition rate the MCA, due to its more expansive structure, was less prone to occlusion than the ACA. The results also showed the effect of a single blockage is expected to be less severe in asymmetric flow than symmetric flow. This model will assist in developing a better understanding into embolic stroke and effect of