Effects of simulation-based practice on focused assessment with sonography for trauma (FAST) window identification, acquisition, and diagnosis.

We compared the effects of simulator-based virtual ultrasound scanning practice with classroom-based ultrasound scanning practice on participants' knowledge of focused assessment with sonography for trauma (FAST) window quadrants and interpretation, and on participants' performance on live patient FAST examinations. Novices with little or no ultrasound training experience received simulation-based practice (n = 24) or classroom-based practice (n = 24). Participants who received simulation-based practice scored significantly higher on interpreting static images of FAST windows. On live patient examinations where participants scanned the right upper quadrant (RUQ), left upper quadrant (LUQ), and suprapubic quadrant of a normal patient and an ascites-positive patient, the classroom-based practice condition had a shorter scan time for the LUQ and a higher number of participants attaining high-quality window on the RUQ (normal patient only) and suprapubic quadrant (positive patient only) and correct window interpretation on the LUQ (normal patient only). Overall, classroom-based practice appeared to promote physical acquisition skills and simulator-based practice appeared to promote window interpretation skills. Accurate window interpretation is critical to identification of blunt abdominal trauma injuries. The simulator used (SonoSimulator) appears promising as a training tool to increase probe time and to increase exposure to FAST windows reflecting various anatomy and disease states.

I-NET: interactive neuro-educational technology to accelerate skill learning.

The learning of a novel task currently rely heavily on conventional classroom instruction with qualitative assessment and observation. Introduction of individualized tutorials with integrated neuroscience-based evaluation techniques could significantly accelerate skill acquisition and provide quantitative evidence of successful training. We have created a suite of adaptive and interactive neuro-educational technologies (I-NET) to increase the pace and efficiency of skill learning. It covers four major themes: 1) Integration of brain monitoring into paced instructional tutorials, 2) Identifying psychophysiological characteristics of expertise using a model population, 3) Developing sensor-based feedback to accelerate novice-to-expert transition, 4) Identifying neurocognitive factors that are predictive of skill acquisition to allow early triage and interventions. We selected rifle marksmanship training as the field of application. Rifle marksmanship is a core skill for the Army and Marine Corps and it involves a combination of classroom instructional learning and field practice involving instantiation of a well-defined set of sensory, motor and cognitive skills. The instrumentation that incorporates the I-NET technologies is called the Adaptive Peak Performance Trainer (APPT). Preliminary analysis of pilot study data for performance data from a novice population that used this device revealed an improved learning trajectory.

Angiopoietin-2 confers Atheroprotection in apoE-/- mice by inhibiting LDL oxidation via nitric oxide.

Atherosclerosis is promoted by a combination of hypercholesterolemia and vascular inflammation. The function of Angiopoietin (Ang)-2, a key regulator of angiogenesis, in the maintenance of large vessels is unknown. A single systemic administration of Ang-2 adenovirus (AdAng-2) to apoE(-/-) mice fed a Western diet significantly reduced atherosclerotic lesion size ( approximately 40%) and oxidized LDL and macrophage content of the plaques. These beneficial effects were abolished by the inhibition of nitric oxide synthase (NOS). In endothelial cells, endothelial NOS activation per se inhibited LDL oxidation and Ang-2 stimulated NO release in a Tie2-dependent manner to decrease LDL oxidation. These findings demonstrate a novel atheroprotective role for Ang-2 when endothelial cell function is compromised and suggest that growth factors, which stimulate NO release without inducing inflammation, could offer atheroprotection.

Endogenous S-nitrosothiols protect against myocardial injury.

Despite substantial evidence that nitric oxide (NO) and/or endogenous S-nitrosothiols (SNOs) exert protective effects in a variety of cardiovascular diseases, the molecular details are largely unknown. Here we show that following left coronary artery ligation, mice with a targeted deletion of the S-nitrosoglutathione reductase gene (GSNOR(-/-)) have reduced myocardial infarct size, preserved ventricular systolic and diastolic function, and maintained tissue oxygenation. These profound physiological effects are associated with increases in myocardial capillary density and S-nitrosylation of the transcription factor hypoxia inducible factor-1alpha (HIF-1alpha) under normoxic conditions. We further show that S-nitrosylated HIF-1alpha binds to the vascular endothelial growth factor (VEGF) gene, thus identifying a role for GSNO in angiogenesis and myocardial protection. These results suggest innovative approaches to modulate angiogenesis and preserve cardiac function.

Successful repair of esophageal injury using an elastin based biomaterial patch.

An esophageal injury with significant tissue loss is very difficult to repair. We conducted an in vivo study to test our elastin based acellular biomaterial patch to repair such defect. The patch was made from porcine aorta, by decellularization and sterilization. Collagen fibers were preserved to retain mechanical strength and enhance cellular in-growth. Ten domestic pigs underwent right thoracotomy. A 2 cm circular defect was made on the distal esophagus, excising half its circumference, and was repaired using the biomaterial patch and sutures. Soon after the procedure, the animals resumed oral feeding. They were followed for clinical status, weight gain, barium studies, and endoscopic studies, and were killed after 6 weeks to 4 months. All ten animals survived long term, with a procedure success rate of 100% (10 of 10). With the exception of one pneumothorax, no complications occurred, and all animals resumed oral feeding and gained weight. Endoscopic studies showed mucosal coverage by 6 weeks, with minimal stricture at the repair site. Excised specimens showed complete mucosal coverage with regeneration of all three layers. Our biomaterial patch can be used safely and reliably for repair of esophageal injury with significant tissue loss when repaired immediately as in our experiment.

Use of a new elastin patch and glue for repair of a major duodenal injury.

Major duodenal injury with significant tissue loss causes high morbidity and mortality. Our new elastin based heterograft combined with small intestinal submucosa (SIS) and biodegradable glue could be used for repair of such defects. Twenty-four domestic pigs were anesthetized and underwent celiotomy. A 2 cm circular defect was created at the second portion of the duodenum with scissors, excising one-half of its circumference. Our elastin patch, combined with SIS, was applied to cover the defect using biodegradable cyanoacrylate glue and a few sutures. It was then covered with omentum. Animals were followed by weight gain, endoscopic evaluation, and upper GI barium studies. After 2-5 months, animals were sacrificed to obtain specimens. One failed in 3 days due to a technical problem, and one failed in 20 days due to an abdominal abscess. The other 22 animals (22/24, 91.7%) did well, gaining weight. Early endoscopic studies (5-14 d) showed an intact patch. Upper GI studies showed varying degrees of stenosis at the repair site at 3-4 months. Sacrifice after 2-5 months showed complete healing of the defect and a dissolved patch. Our new elastin patch material provides a reliable barrier for repair of duodenal injury, and the biodegradable glue provides quick and easy watertight tissue fusion for our patch.

Use of reconstructed small intestine submucosa for urinary tract replacement.

We used reconstructed SIS (ReSIS), a photocrosslinked biomaterial, to create grafts in various shapes and sizes. Sheets of ReSIS were placed in 14 swine to repair bladder defects, and ReSIS tubes were placed in six swine to replace a segment of excised ureter. Histologic analysis of the bladder repair revealed transitional urothelial cells lining the ReSIS by 1 week. After 2 weeks, fibroblasts and mononuclear cells had infiltrated the ReSIS, neovascularization had occurred, and the urothelial lining was more complex, containing multiple cell layers. After 4 weeks, a definite submucosa was present and the ReSIS was starting to degrade. An initial muscular regeneration was demonstrated at 12 weeks. No foreign body reaction, calcification, or sedimentation was noted in any animal. The ureteral implants showed identical histologic changes, without obstruction or leakage of the replaced segment. The ReSIS allowed rapid urothelial regeneration, ingrowth of new blood vessels, and the orderly deposition and organization of new collagen. Our study demonstrates that the photocrosslinking technique used to create larger sheets and tubes of this biomaterial (ReSIS) does not detract from the positive attributes of the SIS and should improve its usefulness in accomplishing larger bladder augmentations and ureter replacements.

Enhanced laser thrombolysis with photomechanical drug delivery: an in vitro study.

BACKGROUND AND OBJECTIVE: Current techniques for laser thrombolysis are limited because they can not completely clear thrombotic occlusions in arteries, typically leaving residual thrombus on the walls of the artery. The objective of this study was to investigate the possibility of using photomechanical drug delivery to enhance laser thrombolysis by delivering drugs into mural thrombus during laser thrombolysis. STUDY DESIGN/MATERIALS AND METHODS: Three experimental protocols were performed in vitro to quantitatively compare the effectiveness of thrombolysis by 1) constant infusion of drug, 2) laser thrombolysis, and 3) photomechanical drug delivery. A fiber-optic flushing catheter delivered drug (a solution of 1 microm fluorescent microspheres) and light ( a 1 micros pulsed dye laser) into a gelatin-based thrombus model. The process of laser-thrombus interaction was visualized using flash photography and the laser-induced pressure waves were measured using an acoustic transducer. RESULTS: Lumen sizes generated by mechanically manipulating the catheter through the thrombus were smaller than those generated by laser ablation. The microspheres could be driven several hundred microns into the mural thrombus. CONCLUSION: Photomechanical drug delivery has potential for enhancement of laser thrombolysis. Two mechanisms seem to be involved in photomechanical drug delivery: 1) mural deposition of the drug at the ablation site and 2) increased exposure of the thrombus surface area to the drug.

Cell damage induced by Angiovist-370 and 308nm excimer laser radiation.

BACKGROUND AND OBJECTIVE: Radiographic contrast media containing iodine-labeled organic compounds can be present in the irradiated field during laser angioplasty using 308 nm excimer laser radiation. These compounds absorb light at 308 nm and may undergo photochemical reactions that produce products that damage cells. STUDY DESIGN/MATERIALS AND METHODS: This study was undertaken to determine whether photoproducts that damage human lymphocytes in vitro are formed when Angiovist 370 (AV), a contrast medium containing triiodinated aromatic compounds, is exposed to 308 nm radiation. RESULTS: The absorption spectrum of AV developed a new peak at 360 nm that extended to wavelengths greater than 500 nm when dilute AV solutions were exposed to 308 nm radiation indicating that photoproducts were formed. Irradiating dilute AV solutions above a layer of human lymphocytes caused a dose-dependent decrease in thymidine incorporation using fluence rates between 5.2 x 10(6) and 1.0 x 10(8) W/cm2. Decreased DNA synthesis was independent of the pulse length (10 ns vs. 230 ns) but was lower, at a given dose, when the highest fluence rate was used. Incubation of lymphocytes with preirradiated AV solutions also decreased incorporation of thymidine in a radiation dose-dependent manner. The cell damaging photoproducts in preirradiated AV solutions were unstable; within 15 min, the effectiveness had decreased by approximately 85%. CONCLUSIONS: These results indicate that exposure of AV to 308 nm excimer laser radiation produces photochemical products that damage human cells in vitro.

Optical mapping in a new guinea pig model of ventricular tachycardia reveals mechanisms for multiple wavelengths in a single reentrant circuit.

BACKGROUND: Although the relationship between cardiac wavelength (lambda) and path length importantly determines the stability of reentrant arrhythmias, the physiological determinants of lambda are poorly understood. To investigate the cellular mechanisms that control lambda during reentry, we developed an experimental system for continuously monitoring lambda within a reentrant circuit with the use of voltage-sensitive dyes and a new guinea pig model of ventricular tachycardia (VT). METHODS AND RESULTS: Action potentials were recorded simultaneously from 128 ventricular sites in Langendorff-perfused hearts (n = 15) in which propagation was confined to a two-dimensional rim of epicardium by an endocardial cryoablating procedure. The reentrant path was precisely controlled by creating an epicardial obstacle (2 x 10 mm) with an argon laser. To control for fiber orientation and rate-dependent membrane properties, lambda during reentry was compared with lambda during plane wave propagation transverse and longitudinal to cardiac fibers at a stimulus cycle length (CL) comparable to the VT CL. Reentrant VT (CL = 97.0 +/- 6.2 ms) was reproducibly induced by programmed stimulation in 93% of preparations. lambda varied considerably within the reentrant circuit (range, 10.6 to 22.5 mm), because of heterogeneities of conduction rather than action potential duration. lambda was significantly shorter during reentrant propagation (ie, with pivoting) parallel to fibers (10.6 +/- 4.2 mm) compared with plane wave propagation (ie, without pivoting) parallel to fibers (32.8 +/- 6.5 mm, P < .02), indicating that wave-front pivoting was primarily responsible for shortening of lambda during reentry. The mechanism of lambda shortening was conduction slowing from increased current load experienced by the pivoting wave front. CONCLUSIONS: We provide direct experimental evidence that multiple wavelengths are present even within a relatively simple reentrant circuit. Abrupt changes in loading during wave-front pivoting, rather than membrane ionic properties or fiber structure, were a major determinant of lambda and, therefore, may play an important role in the stability of reentry.

Drug delivery with microsecond laser pulses into gelatin.

Photo acoustic drug delivery is a technique for localized drug delivery by laser-induced hydrodynamic pressure following cavitation bubble expansion and collapse. Photoacoustic drug delivery was investigated on gelatin-based thrombus models with planar and cylindrical geometries by use of one microsecond laser pulses. Solutions of a hydrophobic dye in mineral oil permitted monitoring of delivered colored oil into clear gelatin-based thrombus models. Cavitation bubble development and photoacoustic drug delivery were visualized with flash photography. This study demonstrated that cavitation is the governing mechanism for photoacoustic drug delivery, and the deepest penetration of colored oil in gels followed the bubble collapse. Spatial distribution measurements revealed that colored oil could be driven a few millimeters into the gels in both axial and radial directions, and the penetration was less than 500 µm when the gelatin structure was not fractured.

Characterization of pulsed-dye laser-mediated vasodilatation in a rabbit femoral artery model of vasoconstriction.

Vasoconstriction is a clinical problem associated with invasive vascular procedures, microvascular reconstruction and subarachnoid hemorrhage. We sought to characterize the ability of pulsed-dye laser irradiation to reverse and prevent vasoconstriction in an anesthetized rabbit model of surgically and pharmacologically induced vasoconstriction. Five groups of experiments were performed to study the effect of pulsed-dye laser irradiation delivered through a 320 microns core ball-tip fiber into the femoral artery. The studies demonstrated that pulsed-dye irradiation can reproducibly cause vascular dilatation. The zone of vasodilatation propagated equally proximal and distal to the site of irradiation within the vessel. When saline was infused into the vessel to replace flowing blood during delivery of laser irradiation, no significant vasodilatation occurred. After laser irradiation reversed surgical and pharmacologic vasoconstriction, the vessel was resistant to further pharmacologic vasoconstriction. This resistance to pharmacologic vasoconstriction did not occur if the vessel was pharmacologically predilated before delivery of laser irradiation. Pathologic analysis of the vessels revealed endothelial damage and mild to moderate medial necrosis, most significant at the site of energy delivery. These studies provide characterization of pulsed-dye laser-mediated vasodilatation in an in vivo model. Delivery of pulsed-dye laser energy has potential clinical application and warrants further investigation.

Treatment of vasospasm with a 480-nm pulsed-dye laser.

Laser energy at a wavelength of 480 nm was applied in 1-microseconds pulses of 3 to 10 mJ to two models of vasospasm. Rabbit common carotid arteries (CCA's) were constricted chronically by the application of human blood within a silicone sheath. Peak vasospasm developed 24 to 48 hours later, and persisted for up to 6 days. Endovascular laser treatment was delivered to 40 CCA's via a 200-microns diameter silica quartz fiber introduced through the femoral artery. The CCA caliber increased from 60% of the pre-vasospasm control diameter to a minimum post-laser diameter of 83% of control. No instances of laser-induced perforation or of arterial thrombosis were observed for up to 60 days after treatment. Prophylactic laser application to nine normal vessels was able to attenuate the development of vasospasm if blood was applied immediately thereafter (88% vs. 59% of control diameter, p less than 0.02), but not if blood was applied 7 days later. Studies in 16 normal CCA's established that there was a considerable margin between the laser energy required to induce dilatation and that which caused perforation, providing that the fiber remained relatively central within the artery. Morphological examination demonstrated focal loss of endothelial cells immediately after laser application, followed approximately 7 days later by the development of areas of intimal hyperplasia. Only minimal changes were observed in the medial or adventitial layers. In a second study, the basilar artery of seven dogs was constricted chronically by two intracisternal injections of autologous blood 3 days apart. Five dogs received endovascular laser treatment 7 or 10 days after the first injection, when basilar artery diameter was reduced to a mean of 61% and 77% of control, respectively. Immediately following treatment, basilar artery diameter increased to 104% and 102% of resting diameter, respectively. Both untreated and laser-treated arteries were smaller than the control diameter at 30 days (80% and 82%, respectively), but in each group the vasodilatory response to hypercapnia was preserved. These findings indicate that 1-microsecond laser pulses are well tolerated by systemic and cerebral arteries in two different animal models, and suggest that the 480-nm pulsed-dye laser may have an application for the treatment or prophylaxis of cerebral vasospasm.

Application of the 1-microsecond pulsed-dye laser to the treatment of experimental cerebral vasospasm.

Laser energy of 480 nm was applied in 1-microsecond pulses varying between 2.2 and 10 mJ to in vitro and in vivo models of cerebral vasospasm. First, the pulsed-dye laser was applied intravascularly via a 320-microns fiber to basilar artery segments from six dogs. The segments were mounted in a vessel-perfusion apparatus and constricted to, on average, 70% of resting diameter by superfusion with dog hemolysate. Immediate increase in basilar artery diameter occurred to a mean of 83% of control. In a second model, the basilar artery was exposed transclivally in the rabbit. In three normal animals, superfusion of the artery with rabbit hemolysate resulted in a reduction of mean vessel diameter to 81% of control. Following extravascular application of the laser, vessels returned to an average of 106% of the resting state. In six rabbits, the basilar artery was constricted by two intracisternal injections of autologous blood. 3 days apart. Two to 4 days after the second injection, the basilar artery was exposed. Extravascular laser treatment from a quartz fiber placed perpendicular to the vessel adventitia resulted in an immediate 53% average increase in caliber to an estimated 107% of control. No reconstriction was observed over a period of up to 5 hours. Morphologically, damage to the arterial wall was slight. This preliminary investigation suggests that the 1-microsecond pulsed-dye laser may be of benefit in the treatment of cerebral vasospasm.

Ultrasound guidance of laser atherectomy.

Intravascular ultrasound imaging catheters and forward firing laser atherectomy systems are both being used in the coronary vascular tree of man. When used in conjunction, ultrasound imaging may reduce the probability of perforation and dissection related to laser atherectomy. A novel system is presented which combines ultrasound imaging and laser atherectomy capabilities on the same catheter. The system incorporates a holmium YAG laser directed perpendicular to the long axis of the catheter along with a co-directional ultrasound sensor. Ultrasound images are assembled by computer using the angle of the catheter tip and its linear advance associated with each ultrasound echo signal. In vivo experimental results are presented.

Effect of blood upon the selective ablation of atherosclerotic plaque with a pulsed dye laser.

Laser angioplasty systems with laser energy preferentially absorbed by atherosclerotic plaque may offer a safe method of plaque removal. This study evaluated the effect of blood upon selective energy absorption using a pulsed dye laser at 480 nm. Intra-arterial laser irradiation of normal rabbit femoral arteries demonstrated a perforation threshold energy with blood perfusion of 13.1 mJ per pulse compared to 87.9 mJ with saline (P less than .0001), indicating a deleterious effect in the presence of blood. An adverse effect upon arterial healing at 3 days was noted in sheep following intra-arterial irradiation during blood but not saline perfusion. Normal and atherosclerotic human aorta ablation thresholds differed significantly (P less than .0002) under saline (plaque: 20 mJ and normal: 120 mJ) but the difference under blood (plaque: 5 mJ and normal: 20 mJ) was not significant. We conclude that absorption of laser energy by blood can reduce the effect of differential absorption by endogenous chromophores in normal and pathologic vascular tissues and, therefore, removal of blood may be a prerequisite for selective ablation of atherosclerotic plaques.

Specificity of the effect of dietary cholesterol on rat liver microsomal 3-hydroxy-3-methylglutaryl-coenzyme a reductase activity.

Dietary cholesterol lowers the activity of rat liver microsomal 3-hydroxy-3-methylglutaryl-CoA reductase without affecting various other liver microsomal enzymes. This is consistent with a specific regulatory mechanism and distinguishes the action of cholesterol on 3-hydroxy-3-methylglutaryl-CoA reductase from that of at least one other stimulus known to affect this enzyme.