Robert S. Keynton

Intimal Hyperplasia and Wall Shear in Arterial Bypass Graft Distal Anastomoses: An In Vivo Model Study

The observation of intimal hyperplasia at bypass graft anastomoses has suggested a potential interaction between local hemodynamics and vascular wall response. Wall shear has been particularly implicated because of its known effects upon the endothelium of normal vessels and, thus, was examined as to its possible role in the development of intimal hyperplasia in arterial bypass graft distal anastomoses. Tapered (4-7 mm I.D.) e-PTFE synthetic grafts 6 cm long were placed as bilateral carotid artery bypasses in six adult, mongrel dogs weighing between 25 and 30 kg with distal anastomotic graft-to-artery diameter ratios (DR) of either 1.0 or 1.5. Immediately following implantation, simultaneous axial velocity measurements were made in the toe and artery floor regions in the plane of the anastomosis at radial increments of 0.35 mm, 0.70 mm, and 1.05 mm using a specially designed 20 MHz triple crystal ultrasonic wall shear rate transducer Mean, peak, and pulse amplitude wall shear rates (WSRs), their absolute values, the spatial and temporal wall shear stress gradients (WSSG), and the oscillatory shear index (OSI) were computed from these velocity measurements. All grafts were harvested after 12 weeks implantation and measurements of the degree of intimal hyperplasia (IH) were made along the toe region and the artery floor of the host artery in 1 mm increments. While some IH occurred along the toe region (8.35+/-23.1 microm) and was significantly different between DR groups (p<0.003), the greatest amount occurred along the artery floor (81.6+/-106.5 microm, mean +/- S.D.) (p < 0.001) although no significant differences were found between DR groups. Linear regressions were performed on the paired IH and mean, peak, and pulse amplitude WSR data as well as the absolute mean, peak, and pulse amplitude WSR data from all grafts. The mean and absolute mean WSRs showed a modest correlation with IH (r = -0.406 and -0.370, respectively) with further improvements seen (r = -0.482 and -0.445, respectively) when using an exponential relationship. The overall best correlation was seen against an exponential function of the OSI (r = 0.600). Although these correlation coefficients were not high, they were found to be statistically significant as evidenced by the large F-statistic obtained. Finally, it was observed that over 75 percent of the IH occurred at or below a mean WSR value of 100 s(-1) while approximately 92 percent of the IH occurred at or below a mean WSR equal to one-half that of the native artery. Therefore, while not being the only factor involved, wall shear (and in particular, oscillators wall shear) appears to provide a stimulus for the development of anastomotic intimal hyperplasia.

Viscous damping of microresonators for gas composition analysis

The damping effect of various gas environments on a silicon, lateral microresonator implemented with piezoresistive detection is investigated in this study. The resonant frequency of the cantilever shifts due to viscous damping by an amount that is directly determined by the molar mass of the gas, thereby providing a method to determine the composition of the gas environment. In addition, the microresonator demonstrates the ability to perform CO2 composition analysis using this nonreaction based detection method. The advantages of this gas analysis method are that it is simple, repeatable, reversible and not limited to reactive gases.

Curcumin Encapsulation in Submicrometer Spray-Dried Chitosan/Tween 20 Particles

Optimal curcumin delivery for medicinal applications requires a drug delivery system that both solubilizes curcumin and prevents degradation. To achieve this, curcumin has been encapsulated in submicrometer chitosan/Tween 20 particles via a benchtop spray-drying process. Spray-drying parameters have been optimized using a Taguchi statistical approach to minimize particle size and to favor spheroid particles with smooth surfaces, as evaluated with scanning electron microscopy (SEM) imaging. Nearly spherical particles with 285 ± 30 nm diameter and 1.21 axial ratio were achieved. Inclusion of curcumin in the spray-drying solution results in complete encapsulation of curcumin within the chitosan/Tween 20 particles. Release studies confirm that curcumin can be released completely from the particles over a 2 h period.

The effect of graft caliber upon wall shear within in vivo distal vascular anastomoses

Wall shear has been widely implicated as a contributing factor in the development of intimal hyperplasia in the anastomoses of chronic arterial bypass grafts. Earlier studies have been restricted to either: (1) in vitro or computer simulation models detailing the complex hemodynamics within an anastomosis without corresponding biological responses, or (2) in vivo models that document biological effects with only approximate wall shear information. Recently, a specially designed pulse ultrasonic Doppler wall shear rate (PUDWSR) measuring device has made it possible to obtain three near-wall velocity measurements nonintrusively within 1.05 mm of the vessel luminal surface from which wall shear rates (WSRs) were derived. It was the purpose of this study to evaluate the effect of graft caliber, a surgically controllable variable, upon local hemodynamics, which, in turn, play an important role in the eventual development of anastomotic hyperplasia. Tapered (4-7 mm I.D.) 6-cm-long grafts were implanted bilaterally in an end-to-side fashion with 30 deg proximal and distal anastomoses to bypass occluded common carotid arteries of 16 canines. The bypass grafts were randomly paired in contralateral vessels and placed such that the graft-to-artery diameter ratio, DR, at the distal anastomosis was either 1.0 or 1.5. For all grafts, the average Re was 432 +/- 112 and the average Womersley parameter, alpha, was 3.59 +/- 0.39 based on artery diameter. There was a sharp skewing of flow toward the artery floor with the development of a stagnation point whose position varied with time (up to two artery diameters) and DR (generally more downstream for DR = 1.0). Mean WSRs along the artery floor for DR = 1.0 and 1.5 were found to range sharply from moderate to high retrograde values (589 s-1 and 1558 s-1, respectively) upstream to high antegrade values (2704 s-1 and 2302 s-1, respectively) immediately downstream of the stagnation point. Although there were no overall differences in mean and peak WSRs between groups, there were significant differences (p < 0.05) in oscillatory WSRs as well as in the absolute normalized mean and peak WSRs between groups. There were also significant differences (p < 0.05) in mean and peak WSRs with respect to axial position along the artery floor for both DR cases. In conclusion, WSR varies widely (1558 s-1 retrograde to 2704 s-1 antegrade) within end-to-side distal graft anastomoses, particularly along the artery floor, and may play a role in the development of intimal hyperplasia through local alteration of mass transport and mechano-signal transduction within the endothelium.

Characterization of micromanipulator-controlled dry spinning of micro- and sub-microscale polymer fibers

No current microfabrication technique exists for producing room- temperature, high-precision, point-to-point polymer micro- and sub- microscale fibers in three dimensions. The purpose of this work is to characterize a novel method for fabricating interconnected three- dimensional (3-D) structures of micron and sub-micron feature size. Poly- (methyl methacrylate) (PMMA) micro- and sub-microscale fiber suspended bridges are fabricated at room temperature by drawing from pools of solvated polymer using a nano-tipped stylus that is precisely positioned by an ultra-high-precision micromilling machine. The fibers were drawn over a 1.8 mm silicon trench, and as the solvent in the solution bridge rapidly evaporates, a suspended, 3-D PMMA fiber remained between the two pools. The resulting fiber diameters were measured for solutions of PMMA in chlorobenzene with concentrations ranging from 15.5 to 23.0 wt% 495k g mol−1 PMMA and 13.0 to 21.0 wt% 950k g mol−1 PMMA. Fibers were found to increase in diameter from 450 nm to 50 µm, roughly corresponding to the increase in concentration of PMMA. To minimize fiber diameter variance, different stylus materials were investigated, with a Parylene®-coated stylus producing fibers with the lowest variance in diameter. Overall, the fiber diameter was found to increase significantly as the solution concentration and polymer molecular weight increased.

Oriented nanomaterial air bridges formed from suspended polymer-composite nanofibers.

In a two-step method, carbon nanotubes, inorganic nanowires, or graphene sheets are connected between two anchor points to form nanomaterial air bridges. First, a recently developed method of forming directionally oriented polymer nanofibers by hand-application is used to form suspended composite polymer-nanomaterial fibers. Then, the polymer is sacrificed by thermally induced depolymerization and vaporization, leaving air bridges of the various materials. Composite fibers and bundles of nanotubes as thin as 10 nm that span 1 microm gaps have been formed by this method. Comparable bridges are observed by electrospinning solutions of the same nanomaterial-polymer composites onto micrometer-scale corrugated surfaces. This method for assembling nanomaterial air-bridges provides a convenient way to suspend nanomaterials for mechanical and other property determinations, and for subsequent device fabrication built up from the suspended nanosubstrates.

Studying Autism Spectrum Disorder with Structural and Diffusion Magnetic Resonance Imaging: A Survey

Magnetic resonance imaging (MRI) modalities have emerged as powerful means that facilitate non-invasive clinical diagnostics of various diseases and abnormalities since their inception in the 1980s. Multiple MRI modalities, such as different types of the sMRI and DTI, have been employed to investigate facets of ASD in order to better understand this complex syndrome. This paper reviews recent applications of structural magnetic resonance imaging (sMRI) and diffusion tensor imaging (DTI), to study autism spectrum disorder (ASD). Main reported findings are sometimes contradictory due to different age ranges, hardware protocols, population types, numbers of participants, and image analysis parameters. The primary anatomical structures, such as amygdalae, cerebrum, and cerebellum, associated with clinical-pathological correlates of ASD are highlighted through successive life stages, from infancy to adulthood. This survey demonstrates the absence of consistent pathology in the brains of autistic children and lack of research investigations in patients under 2 years of age in the literature. The known publications also emphasize advances in data acquisition and analysis, as well as significance of multimodal approaches that combine resting-state, task-evoked, and sMRI measures. Initial results obtained with the sMRI and DTI show good promise toward the early and non-invasive ASD diagnostics.

A new technique for reversible permeabilization of live cells for intracellular delivery of quantum dots.

A major challenge with the use of quantum dots (QDs) for cellular imaging and biomolecular delivery is the attainment of QDs freely dispersed inside the cells. Conventional methods such as endocytosis, lipids based delivery and electroporation are associated with delivery of QDs in vesicles and/or as aggregates that are not monodispersed. In this study, we demonstrate a new technique for reversible permeabilization of cells to enable the introduction of freely dispersed QDs within the cytoplasm. Our approach combines osmosis driven fluid transport into cells achieved by creating a hypotonic environment and reversible permeabilization using low concentrations of cell permeabilization agents like Saponin. Our results confirm that highly efficient endocytosis-free intracellular delivery of QDs can be accomplished using this method. The best results were obtained when the cells were treated with 50 μg ml⁻¹ Saponin in a hypotonic buffer at a 3:2 physiological buffer:DI water ratio for 5 min at 4 °C.

Alzheimer's disease diagnostics by adaptation of 3D convolutional network

Early diagnosis, playing an important role in preventing progress and treating the Alzheimers disease (AD), is based on classification of features extracted from brain images. The features have to accurately capture main AD-related variations of anatomical brain structures, such as, e.g., ventricles size, hippocampus shape, cortical thickness, and brain volume. This paper proposed to predict the AD with a deep 3D convolutional neural network (3D-CNN), which can learn generic features capturing AD biomarkers and adapt to different domain datasets. The 3D-CNN is built upon a 3D convolutional autoencoder, which is pre-trained to capture anatomical shape variations in structural brain MRI scans. Fully connected upper layers of the 3D-CNN are then fine-tuned for each task-specific AD classification. Experiments on the CADDementia MRI dataset with no skull-stripping preprocessing have shown our 3D-CNN outperforms several conventional classifiers by accuracy. Abilities of the 3D-CNN to generalize the features learnt and adapt to other domains have been validated on the ADNI dataset.

Gold Nanoplates as Cancer-Targeted Photothermal Actuators for Drug Delivery and Triggered Release

The selective exposure of cancerous tissue to systemically delivered chemotherapeutic agents remains a major challenge facing cancer therapy. To address this question, a near infrared responsive oligonucleotide-coated AS1411, hairpin, or both gold nanoplate loaded with doxorubicin is demonstrated to be nontoxic to cells without triggered release, while being acutely toxic to cells after 5 minutes of laser exposure to trigger DOX release. Conjugation of oligonucleotides to the nanoplates is confirmed by an average increase in hydrodynamic diameter of 30.6 nm, an average blue shift of the plasmon resonance peak by 36 nm, and an average −10 mV shift in zeta potential of the particles. DOX loading through intercalation into the hairpin DNA structure is confirmed through fluorescence measurements. For both GNP-Hairpin and GNP-Hairpin-AS1411, ~60% of loaded DOX is released after the first 5 minutes of laser exposure λ=817 nm, with complete release after two more 5-minute exposures. Preliminary proof of concept is demonstrated in vitro using A549 and MDA-MB-231 cell lines as models for breast and lung cancer, respectively. Exposure of cells to untriggered DOX-loaded conjugate with no laser exposure results in little to no toxicity, while laser-triggered release of DOX causes significant cell death.