Strain measurement in coronary arteries using intravascular ultrasound and deformable images

Atherosclerotic plaque rupture is responsible for the majority of myocardial infarctions and acute coronary syndromes. Rupture is initiated by mechanical failure of the plaque cap, and thus study of the deformation of the plaque in the artery can elucidate the events that lead to myocardial infarction. Intravascular ultrasound (IVUS) provides high resolution in vitro and in vivo cross-sectional images of blood vessels. To extract the deformation field from sequences of IVUS images, a registration process must be performed to correlate material points between image pairs. The objective of this study was to determine the efficacy of an image registration technique termed Warping to determine strains in plaques and coronary arteries from paired IVUS images representing two different states of deformation. The Warping technique uses pointwise differences in pixel intensities between image pairs to generate a distributed body force that acts to deform a finite element model. The strain distribution estimated by image-based Warping showed excellent agreement with a known forward finite element solution, representing the gold standard, from which the displaced image was created. The Warping technique had a low sensitivity to changes in material parameters or material model and had a low dependency on the noise present in the images. The Warping analysis was also able to produce accurate strain distributions when the constitutive model used for the Warping analysis and the forward analysis was different. The results of this study demonstrate that Warping in conjunction with in vivo IVUS imaging will determine the change in the strain distribution resulting from physiological loading and may be useful as a diagnostic tool for predicting the likelihood of plaque rupture through the determination of the relative stiffness of the plaque constituents.     

Impact of age and hyperglycemia on the mechanical behavior of intact human coronary arteries: an ex vivo intravascular ultrasound study

Despite their advantages, percutaneous coronary interventional procedures are less effective in diabetic patients. Changes in the mechanical properties of vascular walls secondary to long-term hyperglycemia as well as other factors such as age may influence coronary distensibility. This investigation is aimed at deciphering the extent of these effects on distensibility of postmortem human coronary arteries in a controlled manner. Excised human left anterior descending (LAD) coronary arteries were obtained within 24 h postmortem. With the use of intravascular ultrasound, vascular deformation was analyzed at midregions of 51 moderate lesions. Intraluminal pressure was systematically altered using a computerized pressure pump system and monitored by a pressure-sensing guidewire. Distensibility, a normalized compliance term, was defined as the change in lumen area normalized by the initial reference area over a given pressure interval. With the use of multivariate analysis and repeated-measures ANOVA, coronary distensibility was independently influenced by hyperglycemia and age (P < 0.05) through the entire pressure range. Within physiological pressure range, distensibility was significantly reduced with age in nonhyperglycemic coronary specimens (10.55 +/- 4.41 vs. 6.99 +/- 2.45, x10(3) kPa(-1), P = 0.01), whereas the hyperglycemic vessels were stiff even in the younger group (7.90 +/- 5.82 vs. 7.20 +/- 3.36, x10(3) kPa(-1), P = 0.79). Similar results were observed with stiffness index and elastic modulus of the arteries. Hyperglycemia and age independently influenced the distensibility of moderately atherosclerotic LAD coronary arteries. The stiffening with age was overshadowed in the hyperglycemic group by as-yet-undetermined factors.     

Construction of healthy arteries using computed tomography and virtual histology intravascular ultrasound

Vessel geometry for numerical analysis is generally obtained by computed tomography (CT) or magnetic resonance imaging (MRI) and intravascular ultrasound (IVUS). Most medical imaging is obtained from patients for hemodynamic analysis due to the properties of vascular disease and the difficulties in angiography. To predict the site where plaque occurs and understand the progression of the lesion, however, it is necessary to take into consideration not only the diseased artery, but also the blood flow characteristics of healthy artery. In order to simulate healthy vessels prior to lesion formation, we performed CT and virtual histology intravascular ultrasound (VH-IVUS) on three actual patients and this data was used to develop criteria for healthy vessel construction, a method that virtually removes all intravascular plaque. The lumen of a vessel generated by CT and the lumen from VH-IVUS were compared, and the cross-sectional areas of plaque components (fibrous, fibrofatty, dense calcium, and necrotic) and the lumen from VH-IVUS were analyzed. Geometric differences in the healthy vessel and diseased vessel were analyzed, and flow characteristics of the healthy vessel and diseased vessel were compared through computational fluid dynamics simulation. Low average wall shear stress (AWSS) was distributed in the site where plaque was removed from the healthy vessel, and a high oscillatory shear index (OSI) was observed in the region proximal to the site where plaque previously existed. Low AWSS and high OSI are widely accepted indicators of plaque formation or the direction of plaque progression. A numerical model that effectively predicts lesion forming sites was also generated based on the healthy vessel construction method presented in this study.     

A novel intensity-based multi-level classification approach for coronary plaque characterization in intravascular ultrasound images

Background

Intravascular ultrasound (IVUS) is a commonly used diagnostic imaging method for coronary artery disease. Virtual histology (VH) characterizes the plaque components into fibrous tissue (FT), fibro-fatty tissue (FFT), necrotic core (NC), or dense calcium (DC). However, VH can obtain only a single-frame image in one cardiac cycle, and specific software is needed to obtain the radio frequency data. This study proposed a novel intensity-based multi-level classification model for plaque characterization.

Methods

The plaque-containing regions between the intima and the media-adventitia were segmented manually for all IVUS frames. A total of 54 features including first order statistics, grey level co-occurrence matrix, Law’s energy measures, extended grey level run length matrix, intensity, and local binary pattern were estimated from the plaque-containing regions. After feature extraction, optimal features were selected using principle component analysis (PCA), and these were utilized as the input for the classification models. Plaque components were classified into FT, FFT, NC, or DC using an intensity-based multi-level classification model consisting of three different nets. Net 1 differentiated low-intensity components into FT/FFT and NC/DC groups. Then, net 2 subsequently divided FT/FFT into FT or FFT, whereas the remainder and high-intensity components were classified into NC or DC via net 3. To improve classification accuracy, each net utilized three different input features obtained by PCA. Classification performance was evaluated in terms of sensitivity, specificity, accuracy, and receiver operating characteristic curve.

Results

Quantitative results indicated that the proposed method showed significantly high classification accuracy for all tissue types. The classifiers had classification accuracies of 85.1%, 71.9%, and 77.2%, respectively, and the areas under the curve were 0.845, 0.704, and 0.783. In particular, the proposed method achieved relatively high sensitivity (82.0%) and specificity (87.1%) for differentiating between the FT/FFT and NC/DC groups.

Conclusions

These results confirmed the clinical applicability of the proposed approach for IVUS-based tissue characterization.

     

A novel method to estimate the stiffness of the equine back

Diagnosis of back problems in equine orthopedics can be a difficult task. The aim of our study was to develop a new method for estimating the stiffness of the equine back in vivo. We measured the activity of the long back muscle at two locations on both sides at thoracic vertebrae T12 and T16 of 15 horses flexing and extending their back at stance using telemetric surface electromyography, while simultaneously recording the motion of the back with a video camera system. Out of these paired data sets we computed a transfer function in the frequency domain and evaluated its capability of capturing the biomechanical behavior. The transfer function was evaluated via correlation between calculated and actual motion resulting in correlation coefficients of 0.89 for lateral flexion and 0.83 for ventral extension at T16 and 0.82 for lateral flexion and 0.83 for ventral extension at T12. The transfer function was fitted to a filter polynomial of second order, and related to the motion equation. By comparison of coefficients we gained an estimate for the stiffness of the back resulting in a mean value of approximately 6100 N/m for lateral flexion and 650 N/m for ventral extension. This new method enables clinicians in equine orthopedics to estimate back stiffness in horses, and it also provides reality grounded values for biomechanical models of the equine back.     

A novel method to measure cryoprotectant permeation into intact articular cartilage

Successful cryopreservation of articular cartilage (AC) could improve clinical results of osteochondral allografting and provide a useful treatment alternative for large cartilage defects. However, successful cartilage cryopreservation is limited by the time required for cryoprotective agent (CPA) permeation into the matrix and high CPA toxicity. This study describes a novel, practical method to examine the time-dependent permeation of CPAs [dimethyl sulfoxide (DMSO) and propylene glycol (PG)] into intact porcine AC. Dowels of porcine AC (10 mm diameter) were immersed in solutions containing high concentrations of each CPA for different times (0, 15, 30, 60 min, 3, 6, and 24 h) at three temperatures (4, 22, and 37 degrees C), with and without cartilage attachment to bone. The cartilage was isolated and the amount of cryoprotective agent within the matrix was determined. The results demonstrated a sharp rise in the CPA concentration within 15-30 min exposure to DMSO and PG. The concentration plateaued between 3 and 6 h of exposure at a concentration approximately 88-99% of the external concentration (6.8 M). This observation was temperature-dependent with slower permeation at lower temperatures. This study demonstrated the effectiveness of a novel technique to measure CPA permeation into intact AC, and describes permeation kinetics of two common CPAs into intact porcine AC.     

A new method to estimate areas of occupancy using herbarium data

The World Conservation Union (IUCN) Red List of Threatened Species is an important instrument to evaluate the conservation status of living organisms. However, Red List assessors have been limited by the lack of reliable methods to calculate the area of occupancy (AOO) of species, which is an important parameter for red list assessments. Here we present a new practical method to estimate AOO based on herbarium specimen data: the Cartographic method by Conglomerates (CMC). This method, which combines elements from the Areographic and Cartographic methods previously used to calculate AOO, was tested with ten cactus species from the Chihuahuan Desert Region. The results derived from this novel procedure produced in average AOO calculations 3.5 and 5.5 smaller than the Areographic and Cartographic methods, respectively. The CMC takes into account the existence of disjunctions in the distribution range of the species, producing comparatively more accurate AOO estimations. Another advantage of the CMC is that it generates results more harmonic with the current Red List criteria. In contrast, the overestimated results of the Areographic and Cartographic methods tend to artificially categorize the species, even extremely narrow endemics, in lower endangerment status.     

A novel method for the intracellular delivery of siRNA using microbubble-enhanced focused ultrasound

Short interfering RNA (siRNA) has attracted much attention for clinical use in various diseases. However, its delivery, especially through the cell membrane, continues to present a challenge. Advances in ultrasound- and ultrasound contrast-agent technologies have made it possible to change transiently the permeability of the cell membrane and, using a focused ultrasound transducer, to narrow and focus the ultrasound energy on a small target, thereby avoiding damage to surrounding tissue. In this in vitro study, we demonstrate that it is possible to deliver siRNA intracellularly via microbubble-enhanced focused ultrasound. Although further optimization is necessary, our novel method for siRNA transduction represents a powerful tool for using siRNA in vivo and possibly in the clinical setting.     

A novel approach to estimate trabecular bone anisotropy using a database approach

Continuum finite element (FE) models of bones have become a standard pre-clinical tool to estimate bone strength. These models are usually based on clinical CT scans and material properties assigned are chosen as isotropic based only on the density distribution. It has been shown, however, that trabecular bone elastic behavior is best described as orthotropic. Unfortunately, the use of orthotropic models in FE analysis derived from CT scans is hampered by the fact that the measurement of a trabecular orientation (fabric) is not possible from clinical CT images due to the low resolution of such images. In this study, we explore the concept of using a database (DB) of high-resolution bone models to derive the fabric information that is missing in clinical images. The goal of this study was to investigate if models with fabric derived from a relatively small database can already produce more accurate results than isotropic models.     

Tying with thread of the rat coronary arteries, a method for the production of experimental coronary stenosis]

A method is described for producing a localized coronary artery stenosis by tieing two threads around the Ramus descendens of the left coronary artery, which produces progressive cell proliferation of the vascular wall. Studies on 102 male Wistar rats (70 experimental and 32 sham-operated control animals) have shown coronary artery stenosis narrowing the lumen by more than 50% (up to and exceeding 90%) to develop after 7 days in 30% of the animals, after 14 days in 75%, and after 21 days (postoperatively) in 100%. In about one-third of the cases subtotal infarction in the supply area of the left coronary artery occured as a result of post-operative damage to the coronary artery with obturating thrombosis. The induction of effective coronary stenosis limiting the coronary reserve offers possibilities for further experimental studies on the role of relative coronary insufficiency in the pathogenesis of myocardial infarction.     

A method for experimental heating of intact soil profiles for application to climate change experiments

A new system for simulating future belowground temperature increases was conceived, simulated, constructed and tested in a temperate deciduous forest in Oak Ridge, TN, USA. The new system uses low‐wattage, 3 m deep heaters installed around the circumference of a defined soil volume. The heaters add the necessary energy to achieve a set soil temperature differential within the treatment area and add exterior energy inputs equal to those, which might be lost from lateral heat conduction. The method, which was designed to work in conjunction with aboveground heated chambers, requires only two control sensor positions one for aboveground air temperatures at 1 m and another for belowground temperatures at 0.8 m. The method is capable of achieving temperature differentials of at least +4.0±0.5 °C for soils to a measured depth of ?2 m. These +4 °C differential soil temperatures were sustained in situ throughout 2009, and both diurnal and seasonal cycles at all soil depths were retained using this simple heating approach. Measured mean energy inputs required to sustain the target heating level of +4 °C over the 7.1 m² target area were substantial for aboveground heating (21.1 kW h day^?1 m^?2), but 16 times lower for belowground heaters (1.3 kW h day^?1 m^?2). Observations of soil CO₂ efflux from the surface of the target soil volumes showed CO₂ losses throughout 2009 that were elevated above the temperature response curve that have been reported in previous near‐surface soil warming studies. Stimulation of biological activity within previously undisturbed deep‐soil carbon stocks is the hypothesized source. Long‐term research programs may be able to apply this new heating method that captures expected future warming and temperature dynamics throughout the soil profile to address uncertainties in process‐level responses of microbial, plant and animal communities in whole, intact ecosystems.     

利用Inverse PCR快速筛选单个T—DNA拷贝转基因水稻植株的方法

In order to obtain single T-DNA copy transgenic rice, we have established a quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR). IPCR was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences, which will help to estimate the T-DNA copy number in transgenic rice. We have analyzed 20 transgenic plants of 15 transgenic lines. Most plants (12) contain one integrated T-DNA copy per genome, 3 plants contain two and 1 plant contains 3 copies. In 4 transgenic plants no T-DNA copies could be detected using this method. The IPCR results were further tested by Southern analysis and sequence analysis.     

A novel method to identify topological domains using Hi-C data

Over the last decade the 3C-based (Chromosome Conformation Capture, 3C) approaches have been developed to describe the frequency of chromatin interaction. The invention of Hi-C allows us to obtain genome-wide chromatin interaction map. However, it is challenging to develop efficient and robust analytical tools to interpret the Hi-C data. Here we present a new method called Clustering based Hi-C Domain Finder (CHDF), which is based on the difference of interaction intensity inside/outside domains, to identify Hi-C domains. We also compared CHDF with existing methods including Direction Index (DI) and HiCseg. CHDF can define more chromatin domains validated by higher resolution local chromatin structure data (Chromosome Conformation Capture Carbon Copy (5C) data). Using Hi-C data of lower sequencing depth, chromatin structure identified by CHDF is closer to that discovered by data of higher sequencing depth. Furthermore, the implement of CHDF is faster than the other two. Using CHDF, we are potentially able to discover more hints and clues about chromatin structural elements at domain level.     

A novel microsurgery method for intact plant tissue at the single cell level using ArF excimer laser microprojection

A novel microsurgery technique for the partial removal of rigid cell-walls in intact plant tissue is established. Using a size-variable slit, an ArF excimer laser was microprojected on the surface of the targeted cell, and this method enabled the area- and depth-controllable processing of the cortical structure of plant cells including the cuticle and cell wall layer. In epidermal cells of all tested plants, viabilities of more than 90% were retained 24 h after irradiation. Scanning electron microscope (SEM) observation revealed that the cuticle layer of the irradiated region was completely ablated, and the cellulose microfibrils of the secondary cell wall were partially removed; furthermore, 4 days after laser treatment, the regeneration of cell wall fibrils was observed. As a model experiment, the transient expression of synthetic green fluorescent protein (sGFP) was performed by the microinjection of cauliflower mosaic virus (CMV) 35S promoter-derived sGFP gene through an "aperture" in the treated cell surface. Moreover, micron-sized fluorescent beads were successfully introduced by the same method into the onion cells indicating that this method can be used to introduce foreign materials as large as organelles.