Three-dimensional reconstruction of the coronary artery wall by image fusion of intravascular ultrasound and bi-plane angiography.

BACKGROUND: Intravascular ultrasound (IVUS) is becoming increasingly accepted for assessing coronary anatomy. However, its utility in visualizing and quantifying coronary morphology has been limited by its 2D tomographic nature. This study presents a 3D reconstruction technique that accurately preserves 3D geometric information. METHODS AND RESULTS: Images obtained from manual IVUS pullbacks and continuous bi-plane angiography were fused, using angiography to reconstruct the transducer trajectory and aid in solving for the correct rotational orientation. A novel 3D active surface method automatically identified the luminal and medial-adventitial borders which, when superimposed on the transducer trajectory, could be surface-rendered for visualization and morphometry. Segmentation agreed well with manual assessment, and 3D luminal shape matched that of angiography when projected to 2D. CONCLUSIONS: We conclude that this method provides an accurate reconstruction of the vessel's anatomy, which accounts for the true curvature of the vessel.

Assessment of coronary compensatory enlargement by three-dimensional intravascular ultrasound.

Several techniques have been used to demonstrate that human arteries respond to atherosclerosis by increasing their total arterial area to prevent a decrease in blood flow. Three-dimensional reconstructions of coronary arteries can document this compensatory response accurately and specifically. Seven human coronary arteries were reconstructed using intravascular ultrasound and biplane angiography, and vessel geometries were quantified. In all seven vessels, as plaque area increased, overall vessel area increased (R = 0.986, 0.933, 0.984, 0.678, 0.763, 0.963, and 0.830), but luminal cross-sectional area did not significantly decrease. Focal compensatory enlargement was identified in each vessel, and in some cases this response appeared to occur until the vessel was 65% occluded. Luminal enlargement near the proximal ends was attributed to the natural taper of the vessel. The semi-automated, three-dimensional segmentation technique used in this study allows reproducible quantification, as there is no subjective manual tracing involved. Following the intravascular ultrasound transducer in time and space with biplane angiography allows for accurate reconstruction with or without automated pullback devices. Information on the rate of change of vessel measurements is also presented, which, when combined with visualization of accurate 3D geometry, provides a unique assessment of coronary compensatory enlargement. This reconstruction technique can be applied in a clinical environment with no major modification.

Comparison of texture analysis methods for the characterization of coronary plaques in intravascular ultrasound images.

Intravascular Ultrasound (IVUS) is a diagnostic imaging technique that provides tomographic visualization of coronary arteries. The aim of this study was to evaluate five texture analysis techniques and determine their ability to distinguish between plaque lesions of different composition. Using histological correlation, regions of calcified, fibrous, and necrotic core plaque were chosen from 27 coronary plaques. First-order statistics, Haralick's method, Laws' texture energy method, the neighborhood gray-tone difference matrix method, and texture spectrum features were examined using discriminant analysis. Self-validation indicated that Haralick's method yielded the most accurate results, with resubstitution and cross-validation error rates of 0.00 and 14.76%, respectively. Further optimization gave error rates of 6.67%, using only two discriminating features, IDM and entropy.

Vascular mechanics of the coronary artery.

UNLABELLED: This paper describes our research into the vascular mechanics of the coronary artery and plaque. The three sections describe the determination of arterial mechanical properties using intravascular ultrasound (IVUS), a constitutive relation for the arterial wall, and finite element method (FEM) models of the arterial wall and atheroma. METHODS: Inflation testing of porcine left anterior descending coronary arteries was conducted. The changes in the vessel geometry were monitored using IVUS, and intracoronary pressure was recorded using a pressure transducer. The creep and quasistatic stress/strain responses were determined. A Standard Linear Solid (SLS) was modified to reproduce the non-linear elastic behavior of the arterial wall. This Standard Non-linear Solid (SNS) was implemented into an axisymetric thick-walled cylinder numerical model. Finite element analysis models were created for five age groups and four levels of stenosis using the Pathobiological Determinants of Atherosclerosis Youth (PDAY) database. RESULTS: The arteries exhibited non-linear elastic behavior. The total tissue creep strain was epsilon creep = 0.082 +/- 0.018 mm/mm. The numerical model could reproduce both the non-linearity of the porcine data and time dependent behavior of the arterial wall found in the literature with a correlation coefficient of 0.985. Increasing age had a strong positive correlation with the shoulder stress level, (r = 0.95). The 30% stenosis had the highest shoulder stress due to the combination of a fully formed lipid pool and a thin cap. CONCLUSIONS: Studying the solid mechanics of the arterial wall and the atheroma provide important insights into the mechanisms involved in plaque rupture.

Automated identification of stained cells in tissue sections using digital image analysis.

OBJECTIVE: To develop a novel automated image analysis system to differentiate immunohistochemically stained cells from background. STUDY DESIGN: Cell segmentation was performed by applying global thresholding algorithms to find an approximate threshold at which cells could be separated from background followed by a novel refinement algorithm to erode edge pixels of the region. To separate overlapping cells, a new decomposition method was developed that uses both semantic knowledge and high-level relational information. Both the cell segmentation and separation methods were evaluated on images of stained tissue sections and the manually outlined cell areas and numbers compared to the computed. RESULTS: Macrophage areas computed at the first stage by Otsu's algorithm did not differ significantly (P = .07) from those traced manually, while the areas computed by Kittler's and Kurita's algorithms did not agree (P < .01). Both Otsu's and Kurita's algorithms performed well when combined with edge pixel erosion. Kittler's algorithm proved unsuccessful even with edge erosion. Comparison of the computed and manually determined cell numbers showed a significant correlation, and regression analysis resulted in the unity curve. CONCLUSION: A combination of global thresholding and a novel edge erosion technique allowed identification of immunohistochemically stained macrophages; the computed cell areas agreed with the manual results.

Correlation of valve closing to visualization of single-leg separation in radiographic images of Björk-Shiley convexo-concave heart valves.

BACKGROUND AND AIM OF THE STUDY: The study goal was to determine whether the visualization of single-leg separation (SLS) in cineangiographic sequences of Björk-Shiley convexo-concave heart valves could be correlated to the position of the occluder disk within the cardiac cycle. METHODS: Images from ten patient cases with SLS valves were reviewed by three experts, who identified the image frames within a cine sequence that appeared suspicious for SLS. The position of the occluder disk, the frame rate, and the length of the cardiac cycle were noted relative to these image frames. RESULTS: The probability of detecting a SLS was not significantly correlated to any of these factors. CONCLUSIONS: Visualization of SLS in cineangiographic images is limited to a few frames within an imaging sequence. It appears that other features within the image play a larger role in a clinician's ability to detect a fracture than do the cardiac dynamics of the system.

Prevalence and extent of atherosclerosis in adolescents and young adults: implications for prevention from the Pathobiological Determinants of Atherosclerosis in Youth Study.

CONTEXT: Atherosclerosis, the underlying cause of coronary heart disease, has been shown to be present even in young adults. OBJECTIVE: To document the extent and severity of atherosclerosis in adolescents and young adults in the United States. DESIGN AND SETTING: The Pathobiological Determinants of Atherosclerosis in Youth Study, a multi-institutional autopsy study conducted in US medical centers. Subjects A total of 2876 study subjects, between 15 and 34 years old, black and white, men and women, who died of external causes and underwent autopsy between June 1, 1987, and August 31, 1994. MAIN OUTCOME MEASURES: Extent, prevalence, and topography of atherosclerotic lesions. RESULTS: Intimal lesions appeared in all the aortas and more than half of the right coronary arteries of the youngest age group (15-19 years) and increased in prevalence and extent with age through the oldest age group (30-34 years). Fatty streaks were more extensive in black subjects than in white subjects, but raised lesions did not differ between blacks and whites. Raised lesions in the aortas of women and men were similar, but raised lesions in the right coronary arteries of women were less than those of men. The prevalence of total lesions was lower in the right coronary artery than in the aorta, but the proportion of raised lesions among total lesions was higher in the right coronary artery than in the aorta. CONCLUSIONS: Atherosclerosis begins in youth. Fatty streaks and clinically significant raised lesions increase rapidly in prevalence and extent during the 15- to 34-year age span. Primary prevention of atherosclerosis, as contrasted with primary prevention of clinically manifest atherosclerotic disease, must begin in childhood or adolescence.

Three-dimensional segmentation of luminal and adventitial borders in serial intravascular ultrasound images.

Intravascular ultrasound (IVUS) provides exact anatomy of arteries, allowing accurate quantitative analysis. Automated segmentation of IVUS images is a prerequisite for routine quantitative analyses. We present a new three-dimensional (3D) segmentation technique, called active surface segmentation, which detects luminal and adventitial borders in IVUS pullback examinations of coronary arteries. The technique was validated against expert tracings by computing correlation coefficients (range 0.83-0.97) and William's index values (range 0.37-0.66). The technique was statistically accurate, robust to image artifacts, and capable of segmenting a large number of images rapidly. Active surface segmentation enabled geometrically accurate 3D reconstruction and visualization of coronary arteries and volumetric measurements.

A fast and accurate method for registration of MR images of the head.

This paper proposes a new fully automated technique that can be used for the registration of medical images of the head. The method uses Chebyshev polynomials in order to approximate and then minimize a novel multiresolutional, signal intensity independent disparity function, which can generally be defined as the mean squared value of the mean weighted ratio of two images. This function is explicitly computed for n Chebyshev points in a geometric transformation parameter interval [-A, +A] transformation units and is approximated using the Chebyshev polynomials for all other points in the interval. For 3D T2-T1 weighted MR registration, 120 experiments with studies from ten patients were performed and showed that n = 4 Chebyshev points for A = 18 transformation units give mean rotational error 0.36 degrees and a mean translational error 0.36 mm. The different noise conditions did not affect the performance of the method. We conclude that the method is suitable for routine clinical applications and that it has significant potential for future development and improvement.

Relation of a postmortem renal index of hypertension to atherosclerosis and coronary artery size in young men and women. Pathobiological Determinants of Atherosclerosis in Youth (PDAY) Research Group.

In a cooperative multicenter study, the Pathobiological Determinants of Atherosclerosis in Youth, we measured atherosclerosis of the aorta and right coronary artery (RCA) in 2403 black and white men and women 15 through 34 years of age who died of external causes and were autopsied in forensic laboratories. We measured the diameter of the opened, flattened, and fixed RCA and the diameter, intimal thickness, intimal cross-sectional area, medial thickness, and medial cross-sectional area of the pressure-perfused, fixed left anterior descending (LAD) coronary artery. Using the ratio of intimal thickness to outer diameter of the small renal arteries to predict mean arterial pressure during life, we classified the cases as normotensive (mean arterial pressure < 110 mm Hg) or hypertensive (mean arterial pressure > or = 110 mm Hg). The prevalence of hypertension by age, sex, and race corresponded closely with that measured in a survey of the living population. Hypertension had little or no effect on fatty streaks. Hypertension was associated with more extensive raised lesions in the abdominal aortas and RCAs of blacks > 20 years of age and in the RCAs of whites > 25 years of age. At all ages, women had less extensive raised lesions in the RCAs than did men, but the effect of hypertension on raised lesions was similar to that in men. Adjustment for serum lipoprotein cholesterol levels and smoking in a subset of cases yielded results similar to those obtained without adjustment. Hypertension was associated with larger diameters of the RCA and LAD coronary artery and with larger cross-sectional intimal and medial areas of the LAD coronary artery. Hypertension augments atherosclerosis in both men and women primarily by accelerating the conversion of fatty streaks to raised lesions beginning in the third decade of life, and the effect of hypertension increases with age.

Quantification of intramural calcification in coronary intravascular ultrasound images with automated image analysis.

BACKGROUND: Recent studies have documented the utility of intravascular ultrasonography in quantifying coronary morphologic characteristics and determining an appropriate intervention. Unfortunately, its potential for quantifying lesion calcification is limited by subjective evaluation and manual tracing. The aim of this study was to develop an objective automated method for quantifying calcification in intracoronary images with digital image analysis. METHODS: Images of human coronary arteries acquired with a 30 MHz intracoronary ultrasound catheter were evaluated with digital image analysis and compared with manual tracings. Calcifications were automatically identified as highly echogenic regions detected by global thresholding within sectors of acoustic shadowing defined as regions devoid of texture. RESULTS: The mean percentage agreement, sensitivity, and specificity of detecting calcification in 1-degree sectors of calcified vessels were 82%, 73%, and 87%, respectively. Similar results were obtained in noncalcified images. CONCLUSION: The accuracy of this automated technique was comparable to interoperator and intraoperator variability in manually tracing calcification.

Age-related development of atherosclerotic plaque stress: a population-based finite-element analysis.

BACKGROUND: In order to identify those age-related factors in the development of coronary atherosclerosis that would affect the stability of the plaque system, we have developed idealized, finite-element, cross-sectional models of the arterial wall and associated lesions, derived from population-based data. METHODS: The physical development and morphology of coronary plaques was documented in the Pathobiological Determinants of Atherosclerosis in Youth histological study. Using this database, finite-element analysis models were created for five age groups (15-19, 20-24, 25-29 and 30-34 years) and for the 25 largest lesions. Cosmos (Structural Research, Inc., Los Angeles, California, USA) was used to create and analyze the models. RESULTS: The area of greatest stress shifted from the intima opposite the lesion in the 15-19 years age group to the edge of the cap and adjacent healthy tissue in the later age groups. Increasing age had a strong positive correlation with the shoulder stress level (r = 0.95) and the per cent stenosis correlated well with shoulder stress (r = 0.99, P < 0.002). Increasing the cap stiffness from a soft cap to a fibrous cap in the 30-34 year age group model resulted in a localized increase in shoulder surface stress by 10%. A calcified cap increased this shoulder surface stress by 30%. CONCLUSIONS: This finite-element analysis of the population-based data shows that the increase in stress appears to be closely related to the impaired load-bearing capability of the lipid pool that develops with age. The shoulder area of the lesion has been shown to be the location of most of the plaque fractures.

Altered compliance and residual strain precede angiographically detectable early atherosclerosis in low-density lipoprotein receptor deficiency.

BACKGROUND: This study was performed to detect changes in vascular biomechanical properties early in atherogenesis. METHODS AND RESULTS: Age- and weight-matched LDL-receptor deficient Watanabe hypercholesterolemic male rabbits (Group I: n = 11) and normal rabbits (Group II: n = 11) were studied. Fasting plasma lipoprotein concentrations, aortic angiography and intravascular ultrasound, in vivo aortic compliance evaluation, ex vivo aortic residual strain measurements, aortic lipid content and histopathology were determined. Plasma cholesterol was increased 9.8 fold and aortic cholesterol content was increased from 20 to 43 fold in Group I compared to Group II, respectively (P < .00005). Angiography revealed no stenoses in either group, whereas intravascular ultrasound and histological studies of Group I showed small circumferential plaques with < 10% cross-sectional area involvement. The residual strain in Group I was significantly increased in the ascending thoracic aorta (22.1 +/- 6.9% versus 10.4 +/- 3.2% in Group II, P < .0001), descending thoracic aorta (15.7 +/- 7.2% versus 4.8 +/- 1.3% in Group II, P < .0001), and abdominal aorta (18.0 +/- 4.8% versus 8.3 +/- 6.3% in Group II, P < .005). Changes in residual strain were inversely correlated with the aortic cholesterol content in the ascending thoracic aorta (r = -.72; P = -.001), descending thoracic aorta (r = -.95; P < .001), and abdominal aorta (r = -.51; P = .019). CONCLUSIONS: Early atherosclerosis in LDL-receptor deficient rabbits, undetectable by angiography yet observed by intravascular ultrasound imaging and histology, is associated with marked changes in ex vivo residual strain. Alterations in vascular biomechanical properties, associated with changes in cholesterol content, may have physiologic consequences and may be useful in detecting and quantitating early atherosclerosis.

Technical and economic feasibility of reusing disposable perfusion cannulas.

OBJECTIVE(S): The reuse of disposable devices is a potential source of significant cost savings to hospitals. Venous and arterial perfusion cannulas under new and reused conditions were selected to identify the clinical, safety, technical, logistic, and economic issues that must be addressed to realize these savings. METHODS: Single- and dual-stage venous and arterial cannulas from two manufacturers were tested when new, after initial clinical use, and after a single clinical use plus up to nine simulated reuses. Reuse was simulated by end-to-end bending, coupling and uncoupling of the connectors, and by two 1-hour soaks in plasma at 4 degrees and 40 degrees C, respectively. Cannulas were decontaminated and then processed by a peracetic acid-based liquid chemical sterilization system after each use/reuse. Sterilization was validated by eliminating Bacillus subtilis spores from the cannulas on each of five consecutive cycles. Cannulas were tested for physical changes, functional integrity, biocompatibility, and in vivo performance in sheep. A cost analysis was also performed. RESULTS: Sterilization was successfully achieved. Mechanical changes were less than 20% on all variables studied and were undetectable by experienced cardiac surgeons in selective evaluation. No clinically important differences were found between new and reused cannulas, even after nine simulated reuses. Reusing cannulas four times would reduce the cost per procedure from $53 to $19 (64%). CONCLUSIONS: Preliminary data suggest that the perfusion cannulas tested can be safely and efficaciously used five times. Limited reuse of these disposable cannulas is technically feasible and cost-effective. Cannula reuse would result in a small incremental savings; however, with more expensive devices and higher-volume sterilization procedures, the savings could be considerably greater. This program provides a model for evaluation of other single-use medical devices for reuse.

Automated morphometry of coronary arteries with digital image analysis of intravascular ultrasound.

We designed and tested digital image processing strategies to perform fully automated segmentation of luminal and medial-adventitial boundaries in intravascular ultrasound images of human coronary arteries. Automated segmentation is an essential tool for advanced techniques of clinical visualization and quantitative measurement. Vascular compliance measurements and three-dimensional reconstructions are demonstrated as examples of such applications. Digital image processing was performed in three phases: (1) preprocessing, including a polar transform, local contrast enhancement, and speckle noise filtering; (2) segmentation, involving radial scanning, region growing, or cost-function minimization techniques; and (3) postprocessing, involving dropout filtering and outline smoothing. Cross-sectional areas were compared with manual tracings from experienced operators and showed good agreement. The algorithm bias ranged from -0.34 to 1.18 mm2; interclass and intraclass correlation coefficients ranged from 0.83 to 0.94. The designed techniques currently allow fully automated segmentation without operator interaction of the luminal and, if present, medial-adventitial boundary.

Technical and economic feasibility of reusing disposable perfusion cannulae.

The reuse of disposable devices is a potential source of significant cost savings to hospitals. Venous and arterial perfusion cannulae under new and reused conditions were selected to identify the clinical, safety, technical, logistic, and economic issues that must be addressed to realize these savings. Single- and dual-stage venous and arterial cannulae from two manufacturers were tested when new, after initial clinical use, and after a single clinical use plus up to nine simulated reuses. Reuse was simulated by end-to-end bending, coupling and uncoupling the connectors, and by two 1-hour soaks in plasma at 4 degrees C and 40 degrees C, respectively. Cannulae were decontaminated and then sterilized by a peracetic acid based liquid chemical sterilization system following each use/reuse. Sterilization was validated by eliminating Bacillus subtilis spores from the cannulae on each of five consecutive cycles. Cannulae were tested for physical changes, functional integrity, biocompatibility, and in vivo performance in sheep. A cost minimization analysis was also performed. No clinically important differences were found between new and reused cannulae, even after nine simulated reuses. Mechanical changes were less than 20% on all variables studied and were undetectable by experienced cardiac surgeons in selective evaluation. Sterilization was successfully achieved. Reusing cannulae for times would reduce the cost per procedure from $53 to $19 (64%). Perfusion cannulae tested can be safely and efficaciously used five times. This study suggests that reuse would result in a small incremental savings; however, with more expensive devices and higher-volume sterilization procedures, the savings could be exponentially greater. Although this study demonstrates that it may be technically feasible and cost-effective to reuse disposable cannulae, the U.S. Food and Drug Administration does not sanction the reuse of disposable cannulae.

The weighted-distance scheme: a globally optimizing projection ordering method for ART.

The order in which the projections are applied in the algebraic reconstruction technique (ART) has a great effect on speed of convergence, accuracy, and the amount of noise-like artifacts in the reconstructed image. In this paper, a new projection ordering scheme for ART is presented: the weighted-distance scheme (WDS). It heuristically optimizes the angular distance of a newly selected projection with respect to an extended sequence of previously applied projections. This sequence of influential projections may incorporate the complete set of all previously applied projections or any limited time interval subset thereof. The selection algorithm results in uniform sampling of the projection access space, minimizing correlation in the projection sequence. This produces more accurate images with less noise-like artifacts than previously suggested projection ordering schemes.

Mechanical response of an artery using a standard nonlinear solid.

INTRODUCTION: The stress and deformation in an artery are determined from an axisymmetric analysis of thick-walled cylinder with a time-dependent internal pressure. The purpose is to understand how different constitutive, loading, and geometric conditions affect the stress and deformation state within the artery. MATERIALS AND METHODS: The equilibrium, compatibility, and constitutive equations are applied at N discretized points in the arterial wall, in an efficient scheme using Mathcad software on a desktop computer. The constitutive equations are a modification to a standard linear solid, so that one of the linear elements is nonlinear, and so that the 3-D response is anisotropic and dissipates energy only under deviatoric (shearing) stress states. Solution at each successive time increment requires inversion of a 6N by 6N matrix. RESULTS: The model reproduces experimental stress relaxation data with a correlation coefficient of 0.985 and can reproduce quasi-static stress strain data with equal accuracy. Features such as conditioning of the tissue are understood in terms of the time-dependent properties of the tissue. CONCLUSION: The program can produce transient and steady-state responses that closely mimic tissue response. The analysis allows for quick and stable determination of the stress and strain states for a variety of loading conditions.

Quantitative comparison of immunohistochemical staining intensity in tissues fixed in formalin and Histochoice.

Formaldehyde fixatives have traditionally been used to preserve tissues as they impart excellent morphological preservation. Formaldehyde fixes tissue by cross linking, a process which can reduce the antigenicity of tissue and weakens the intensity of immunohistochemical stains. Preliminary studies have shown that Histochoice tissue fixative offers equal or greater staining intensity than neutral buffered formalin (NBF). This study compares these fixatives quantitatively and presents the results in unambiguous statistical terms. Tissue samples were collected, bisected, and fixed in NBF or Histochoice. The sections were stained with a total of 21 antibodies, and color images were collected. The hue, saturation, and value were determined for each positive pixel and an ANOVA performed. Small differences in hue were noted in 8 of 21 cases. Histochoice-fixed tissue gave a greater mean saturation than NBF with 57.1% of the antibodies tested. No significant difference in the saturation was detected in 28.6% of the cases; NBF gave higher mean saturation levels with only 14.3% of the antibodies. Histochoice-fixed tissue was found to give lower values in 66.7% of cases than those prepared with NBF, indicating darker staining. These results show that Histochoice produces staining intensity that is comparable, and in many cases superior, to formalin.

Hypercholesterolemia in the rabbit induced by feeding graded amounts of low-level cholesterol. Methodological considerations regarding individual variability in response to dietary cholesterol and development of lesion type.

While a number of studies have presented detailed examinations of lesion development in the cholesterol-fed rabbit, individual variability in response to cholesterol feeding and type of lesion produced relative to the degree of cholesterol exposure is not well defined. This study analyzed such critical parameters in an attempt to further characterize the model and establish a baseline for future testing of treatments targeted at limiting atherosclerosis. For these experiments, male New Zealand White rabbits were fed atherogenic diets consisting of 0.05%, 0.10%, 0.15%, 0.20%, or 0.25% cholesterol dissolved in 6% peanut oil for 31 to 32 weeks. Raising dietary cholesterol from 0.05% to 0.15% resulted in a less than twofold stepwise increase in total plasma cholesterol (TPC) exposure (area under plasma cholesterol versus time curve), whereas further increases in cholesterol intake resulted in an exponential four- to fivefold increase in TPC exposure. Regression analysis of TPC exposure with aortic sudanophilia demonstrated a threshold of approximately 5000 cholesterol weeks; below this limit lesions were minimal, and above this value the degree of plaque correlated with TPC exposure. Furthermore, a wide biological variability occurred among rabbits with respect to individual responsiveness to dietary cholesterol. In the aorta, various types of plaques, from fatty streaks to atheromatous lesions, were observed, depending on the degree of cholesterol intake. Diets consisting of < 0.15% cholesterol resulted in the development of fatty streak lesions, while transitional lesions and atheromatous plaques were mostly found with higher cholesterol feeding. Coronary artery atherosclerosis was present in > 50% of animals fed diets > or = 0.15% cholesterol. Despite the level of TPC exposure, coronary lesions in epicardial vessels were generally the fibrous type, whereas intramyocardial arteries demonstrated predominantly intimal foam cells. In conclusion, by adjusting dietary cholesterol intake and selecting rabbits with a similar responsiveness to cholesterol, the overall cholesterol exposure can be more closely controlled to minimize the inherent individual variability among animals in this model. The nature of the target lesion must also be carefully considered, because the efficacy of some treatments may depend on the type of atherosclerotic plaque.