Applicability of an entry flow model of the brachial artery for flow models of the hemodialysis fistula.

The native arteriovenous fistula creates a shunt that provides the high blood flow that is needed for dialysis. Lumped parameter hemodynamic models of the arteriovenous fistula can be used to predict shear stresses and pressure losses and can be applied to help understand unsolved problems such as the high rate of arteriovenous fistula maturation failure. These models combine together flow components, such as arteries, stenosis, anastomoses, arterial compliance, and blood inertia, and each component must be modeled with an appropriate pressure-flow relationship. Poiseuille flow is generally assumed for straight vessels, but the unique high flow rates within the brachial artery of an arteriovenous fistula are expected to induce entry flow effects that are neglected in this model. To estimate the importance of these effects, brachial artery flow was modeled in a low-resistance network, such as the one that occurs when an arteriovenous fistula is constructed, through the lumped parameter model, and the predicted flow rates and pressures were compared to those predicted by computational fluid dynamics. When Poiseuille flow was assumed, the flow rate from the lumped parameter model was consistently larger than that from computational fluid dynamics, with a cycle-averaged error of 36.8%. When an entry flow model (Shah) was assumed, the lumped parameter-based flow was 6% lower than the computational fluid dynamics model at the peak of the flow waveform, and the cycle-averaged error was reduced to 7.8%. Thus, in a low-resistance (high flow) arteriovenous fistula circuit, an entry flow model can account for steeper near-wall velocity gradients. This result can provide a useful guide for designing engineering models of the arteriovenous fistula.

Gas-phase detection of solid-state fission product complexes for post-detonation nuclear forensic analysis.

This study presents the first known detection of fission products commonly found in post-detonation nuclear debris samples using solid sample introduction and a uniquely coupled gas chromatography inductively-coupled plasma time-of-flight mass spectrometer. Rare earth oxides were chemically altered to incorporate a ligand that enhances the volatility of the samples. These samples were injected (as solids) into the aforementioned instrument and detected for the first time. Repeatable results indicate the validity of the methodology, and this capability, when refined, will prove to be a valuable asset for rapid post-detonation nuclear forensic analysis.

A novel dynamic layer-by-layer assembled nano-scale biointerface: functionality tests with platelet adhesion and aggregate morphology influenced by adenosine diphosphate.

An improved biointerface was developed, dynamic layer-by-layer self-assembly surface (d-LbL), and utilized as a biologically-active substrate for platelet adhesion and aggregation. Possible clinical applications for this research include improved anti-coagulation surfaces. This work demonstrated the functionality of d-LbL biointerfaces in the presence of platelet-rich-plasma (PRP) with the addition of 20 µM adenosine diphosphate (ADP), a thrombus activator. The surface morphology of the experimental control, plain PRP, was compared to PRP containing additional ADP (PRP + ADP) and resulted in an expected increase of platelet adhesions along the fibrinogen d-LbL substrate. The d-LbL process was used to coat glass slides with fibrinogen, Poly (sodium 4-styrene-sulfonate), and Poly (diallydimethlyammonium chloride). Slides were exposed to PRP under flow and static conditions with and without 20 µM of ADP. Fluorescence microscopy (FM), phase contrast microscopy (PCM), atomic force microscopy (AFM), and field emission-scanning electron microscopy (FE-SEM) were used to evaluate platelet adhesions under the influence of varied shear conditions. PCM images illustrated differences between the standard LbL and d-LbL substrates. FM images provided percent surface coverage values. For high-shear conditions, percent surface coverage values increased when using ADP whereas plain PRP exposure displayed no significant increase. AFM scans also displayed higher mean peak height values and unique surface characteristics for PRP + ADP as opposed to plain PRP. FE-SEM images revealed platelet adhesions along the biointerface and unique characteristics of the d-LbL surface. In conclusion, PRP + ADP was more effective at increasing platelet aggregation, especially under high shear conditions, providing further validation of the improved biointerface.

Basic optics of effect materials.

Effect materials derive their color and effect primarily from thin-film interference. Effect materials have evolved over the decades from simple guanine crystals to the complex multilayer optical structures of today. The development of new complex effect materials requires an understanding of the optics of effect materials. Such an understanding would also benefit the cosmetic formulator as these new effect materials are introduced. The root of this understanding begins with basic optics. This paper covers the nature of light, interference of waves, thin-film interference, color from interference, and color travel.

In vitro verification of multiple-receiver Doppler ultrasound for velocity estimation improvement.

The coherent scattering effect, which introduces noise in Doppler-derived velocity estimates, is caused by constructive and destructive interference of sound waves scattered from multiple particles. Because the phase relationship between signals scattered from different particles depends on the orientation of the receiver, the error in a given velocity estimate depends on the receiver location. To examine this dependence, the velocity of a steady uniform flow was measured simultaneously with a transceiver and three receivers, and the cross-correlation coefficients between velocity estimates for pairs of crystals were calculated. The velocity estimates were nearly independent, with cross-correlation coefficients of approximately 0.2. This result agrees with our previously published numerical simulation studies which demonstrated that the coherent scattering noise in receivers separated by 5 degrees or more was nearly uncorrelated. Consequently, the contribution of coherent scattering noise can be reduced by averaging out noise in signals obtained from multiple receivers.

Influence of arterial elasticity and vessel dilatation on arteriovenous fistula maturation: a prospective cohort study.

BACKGROUND: Arteriovenous fistula maturation requires dilatation of the anastomosed artery and vein. The factors that affect dilatation and the mechanisms by which dilatation promotes maturation are not understood. This pilot study tested two hypotheses: that low arterial elasticity is associated with maturation failure, and that vessel dilatation is required for adequate fistula blood flow during dialysis. METHODS: Thirty-two patients underwent preoperative measurement of small artery elasticity index, and pre-anastomosis measurement of artery and vein luminal diameters during fistula surgery. Fistulas were considered mature if they were used successfully in three consecutive treatments within 6 months. A mathematical model was used to determine whether vessel dilatation is needed for adequate fistula flow. RESULTS: Six fistulas were excluded from analysis of maturation because dialysis did not begin within 6 months. Twenty-one of the remaining 26 fistulas were located in the upper arm. Six of 26 failed to mature, and all 6 developed stenosis. The average small artery elasticity index was lower in failed than in matured fistulas (2.25 versus 3.71 ml/ mmHg x 100, P = 0.02). Artery and vein diameters of the 32 patients ranged from 2.5 to 5.0 and 3.5 to 7.0 mm, respectively. When the diameters were applied to the mathematical model, predicted fistula flows ranged from 412 to 1380 ml/min. CONCLUSIONS: Low arterial elasticity is associated with stenosis and fistula maturation failure. However, vessel dilatation is not needed for adequate blood flow except at the smaller diameters in this study. We speculate that low elasticity promotes development of stenosis. Larger studies are needed to confirm these promising results and to determine whether therapies directed at improving elasticity can improve maturation.

Inflow stenosis obscures recognition of outflow stenosis by dialysis venous pressure: analysis by a mathematical model.

BACKGROUND: Recent studies have shown that inflow stenosis of haemodialysis grafts is more common than previously realized. The influence of inflow stenosis on graft haemodynamics and venous pressure (VP) surveillance has not been previously systematically studied. METHODS: We used a well-established mathematical model to determine the relation between inflow stenosis and static VP (adjusted for mean arterial pressure, VP/MAP), outflow stenosis and artery and vein luminal diameters. We applied low, median and high ratios of artery/vein diameters from 94 patients with grafts. The median ratio was 0.77, indicating that the artery was generally narrower than the vein. RESULTS: The model shows that inflow stenosis reduces VP/MAP. More importantly, however, as outflow stenosis progresses, fixed inflow stenosis causes a delayed increase in VP/MAP followed by a rapid increase at critical outflow stenosis. When both stenoses progress together, their relative rates determine whether and how rapidly VP/MAP increases. The increase in VP/MAP is remarkably abrupt when the rate of inflow stenosis approaches that of outflow stenosis. No increase occurs when inflow stenosis progresses as fast or faster than outflow stenosis. CONCLUSION: Inflow stenosis exerts its most important haemodynamic effect through its interaction with outflow stenosis. As outflow stenosis progresses, inflow stenosis causes a delayed and then rapid increase in VP/MAP at critical outflow stenosis. This increase may not be detected before thrombosis unless measurements are very frequent. Inflow stenosis has an important impact on graft haemodynamics and surveillance because of its location in the relatively narrow inflow tract.

Mathematical model demonstrates influence of luminal diameters on venous pressure surveillance.

BACKGROUND: The reliability of dialysis venous pressure (VP) in detecting stenosis is controversial. A mathematical model may help to resolve the controversy by providing insight into the factors that influence static VP. DESIGN, SETTING, PARTICIPANTS, AND MEASUREMENTS: This study used inflow artery and outflow vein luminal diameters from duplex ultrasound studies of 94 patients. These diameters were applied to a mathematical model, and how they affect the relation among VP, mean arterial pressure (MAP), blood flow, and stenosis was determined. Whether VP/MAP is a valid adjustment for the influence of MAP on VP, and whether the standard VP/MAP referral threshold of 0.50 is valid, were also determined. RESULTS: It was found that there is an approximate one-to-one relation between MAP and VP, so VP/MAP is a valid adjustment. Also, the 0.50 threshold successfully identifies most grafts with stenosis of 65% or more. However, the ratio of artery/vein diameters varied widely between patients, and the ratio independently influences VP/MAP. When the inflow artery is relatively narrow, the VP/MAP increase is delayed followed by a more rapid increase as critical stenosis is reached. CONCLUSIONS: VP/MAP is a valid adjustment for the influence of MAP on VP, and the standard VP/MAP threshold of 0.50 warns of the transition to critical stenosis. However, relatively narrow arteries cause a delay followed by a rapid increase in VP/MAP that may not be detected before thrombosis unless measurements are very frequent. Clinical trials that emphasize trend analysis with frequent measurements are needed to evaluate the efficacy of VP surveillance.

Progress toward the culture and transformation of chicken blastodermal cells.

Chicken blastodermal cells can be cultured for short periods of time and retain the ability to contribute to somatic and germline tissues when injected into gamma-irradiated stage X embryos. Such a method has yet to yield a germline transgenic bird, in part due to the low rate of transgene integration into the avian genome. In addition, the short culture period precludes the identification and expansion of those cells that carry an integrated transgene. In this study, two methods were developed that produced blastodermal cells isolated from stage X Barred Plymouth Rock embryos bearing an integrated transgene. Addition of chick embryo extract to the culture medium enabled expansion of single colonies for multiple passages. Southern blot analysis indicated that the transgenes had integrated as a single copy in most of the clones. Cells from passaged, transgenic embryo cells were injected into irradiated stage X White Leghorn embryos, producing hatched chicks that bore the donor cells in their somatic tissues. Transgene sequences were detected in sperm DNA; however, breeding of chimeras did not result in germline transmission of the transgene, indicating that the contribution of the transgenic cells to the germline was either nonexistent or very low.

Influence of luminal diameters on flow surveillance of hemodialysis grafts: insights from a mathematical model.

Randomized controlled trials have not shown that surveillance of graft blood flow (Q) prolongs graft life. Because luminal diameters affect flow resistance, this study examined whether the influence of diameters on Q can explain the limitations of surveillance. Inflow artery and outflow vein diameters were determined from duplex ultrasound studies of 94 patients. These diameters were applied to a mathematical model for determination of how they affect the relation between Q and stenosis. Also determined was the correlation between Q (by ultrasound dilution) and diameters, stenosis, and mean arterial pressure in 88 patients. Artery and vein diameters varied widely between patients, but arteries generally were narrower than veins. The model predicts that the relation between Q and stenosis is sigmoid: as stenosis progresses, Q initially remains unchanged but then rapidly decreases. A narrower artery increases flow resistance, causing a longer delay followed by a more rapid reduction in Q. In a multiple regression analysis of data from patients, Q correlated with artery and vein diameters, sum of largest stenoses from each circuit segment, and mean arterial pressure (R = 0.689, P < 0.001). This study helps to explain why Q surveillance predicts thrombosis in some patients but not others. Luminal diameters control the relation between Q and stenosis, and these diameters vary widely. During progressive stenosis, the delay and then rapid reduction in Q may impair recognition of low Q before thrombosis occurs. Surveillance outcomes might be improved by taking frequent measurements so that there is no delay in discovering that Q has decreased.

Mathematical model for pressure losses in the hemodialysis graft vascular circuit.

Stenosis-induced thrombosis and abandonment of the hemodialysis synthetic graft is an important cause of morbidity and mortality. The graft vascular circuit is a unique low-resistance shunt that has not yet been systematically evaluated. In this study, we developed a mathematical model of this circuit. Pressure losses (deltaPs) were measured in an in vitro experimental apparatus and compared with losses predicted by equations from the engineering literature. We considered the inflow artery, arterial and venous anastomoses, graft, stenosis, and outflow vein. We found significant differences between equations and experimental results, and attributed these differences to the transitional nature of the flow. Adjustment of the equations led to good agreement with experimental data. The resulting mathematical model predicts relations between stenosis, blood flow, intragraft pressure, and important clinical variables such as mean arterial blood pressure and hematocrit. Application of the model should improve understanding of the hemodynamics of the stenotic graft vascular circuit.

Thresholds for significant decrease in hemodialysis access blood flow.

During hemodialysis access surveillance, referral for evaluation and correction of stenosis is based upon determination that a significant decrease in blood flow (Q) has occurred. However, criteria for determining when a decrease is statistically significant have not yet been established. In this study we established such criteria by analyzing Q variation with the glucose pump test (GPT). We took nine Q measurements in each of 25 patients (18 grafts, 7 fistulas) during three dialysis sessions within a 2-week period (predialysis and during hours 1 and 3). We determined thresholds that define a significant percentage decrease in Q (deltaQ) for various p values. In order to confirm the general applicability of these thresholds, we computed the average within-patient Q variation during the three sessions (computed as a coefficient of variation and referred to as short-term variation). We then determined the relative influences of biological (true) variation and analytical error on short-term variation. We found that deltaQ must be > 33% to be significant at p < 0.05, whereas the threshold is > 17% for p < 0.20. Measuring Q at uniform versus different times during the sessions did not significantly reduce these thresholds. We also found that biological variation was nearly as large as short-term Q variation, whereas analytical error contributed minimally to short-term variation. In conclusion, this study defines thresholds for a significant deltaQ that have wide application in determining access referral for evaluation and correction of stenosis. Selection of a particular threshold should consider the relative importance of avoiding thrombosis versus avoiding unnecessary procedures. If avoiding unnecessary procedures is a priority, then we recommend a threshold of > 33%. These thresholds apply to other methods of measuring Q, provided analytical error is significantly less than biological variation.

In vivo validation of glucose pump test for measurement of hemodialysis access flow.

BACKGROUND: The glucose pump test (GPT) is a recently introduced method of measuring hemodialysis access blood flow (Qa). A validation of GPT during dialysis has not yet been done, and performance characteristics of the method have not yet been fully analyzed. METHODS: The authors studied 33 patients (25 synthetic grafts, 8 autogenous arteriovenous fistulae). Qa measurements by ultrasound dilution (UD) and GPT were done in triplicate during dialysis. In GPT, a baseline blood sample (C(1)) was obtained, followed by infusion of a 10% glucose solution (C(i)) through the arterial needle into the access at 16 mL/min (Q(i)). After 11 seconds, a downstream blood sample (C(2)) was aspirated from the venous needle. C(1) and C(2) glucose were measured by glucometer. Qa was computed by the equation: Qa = Q(i)(C(i) - C(2))/(C(2) - C(1)). A model of the access vascular circuit was used to determine the influence of C(2) aspiration on the Qa measurement. RESULTS: Mean Qa was 1413 mL/min by UD versus 1,496 mL/min by GPT (P = 0.11). There was a strong linear correlation between the 2 methods (r = 0.905; P <0.001). The pooled coefficient of variation was 6.4% for UD and 9.6% for GPT. The circuit model showed that aspiration of C(2) causes an increase in Qa (DeltaQa) that depends on the aspiration rate (Q(ASP)) and fraction of resistance in the circuit that is downstream to the venous needle: DeltaQa = Q(ASP)(Downstream resistance)/(Total resistance). The model predicts the overestimate is approximately 62 mL/min for grafts and 120 mL/min for fistulae but may vary depending on the balance of resistances upstream and downstream to the venous needle. CONCLUSION: This study shows that GPT closely correlates with UD, and the method has adequate precision. GPT is an inexpensive method that may help make Qa measurements more widely available than previously possible.

Biocompatibility of layer-by-layer self-assembled nanofilm on silicone rubber for neurons.

Electrostatic layer-by-layer (LbL) self-assembly, a novel method for ultrathin film coating has been applied to silicone rubber to encourage nerve cell adhesion. The surfaces studied consisted of precursor layers, with alternating cationic poly(ethyleneimine) (PEI) and anionic sodium poly(styrenesulfonate) (PSS) followed by alternating laminin and poly-D-lysine (PDL) layers or fibronectin and PDL layers. Film growth increased linearly with the number of layers. Every fibronectin/PDL and laminin/PDL bilayer was 4.4 and 3.5 nm thick, respectively. All layers were more hydrophilic than the unmodified silicone rubber surface, as determined from contact angle measurements. Of the coatings studied, a PDL layer was the most hydrophilic. A multilayer film with composition [PSS/PEI]3+[fibronectin/PDL]4 or [PSS/PEI]3+[laminin/PDL]4 was highly favorable for neuron adhesion, in contrast to bare silicone rubber substrate. The film coated on silicone rubber is biocompatible for cerebellar neurons with active viability, as shown by lactate dehydrogenase (LDH) assay and fluorescence cellular metabolism observations. These results demonstrate that LbL self-assembly provides an effective approach to apply films with nanometer thickness to silicone rubber. Such only few nanometer thick films are biocompatible with neurons, and may be used to coat devises for long-term implant in the central nervous system.

Biomedical applications of electrostatic layer-by-layer nano-assembly of polymers, enzymes, and nanoparticles.

The introduction of electrostatic layer-by-layer (LbL) self-assembly has shown broad biomedical applications in thin film coating, micropatterning, nanobioreactors, artificial cells, and drug delivery systems. Multiple assembly polyelectrolytes and proteins are based on electrostatic interaction between oppositely charged layers. The film architecture is precisely designed and can be controlled to 1-nm precision with a range from 5 to 1000 nm. Thin films can be deposited on any surface including many widely used biomaterials. Microencapsulation of micro/nanotemplates with multilayers enabled cell surface modification, controlled drug release, hollow shell formation, and nanobioreactors. Both in vitro and in vivo studies indicate potential applications in biology, pharmaceutics, medicine, and other biomedical areas.

Coating and selective deposition of nanofilm on silicone rubber for cell adhesion and growth.

A recently developed method for surface modification, layer-by-layer (LbL) assembly, has been applied to silicone, and its ability to encourage endothelial cell growth and control cell growth patterns has been examined. The surfaces studied consisted of a precursor, with alternating cationic polyethyleneimine (PEI) and anionic sodium polystyrene sulfonate (PSS) layers followed by alternating gelatin and poly-D-lysine (PDL) layers. Film growth increased linearly with the number of layers. Each PSS/PEI bilayer was 3 nm thick, and each gelatin/PDL bilayer was 5 nm thick. All layers were more hydrophilic than the unmodified silicone rubber surface, as determined from contact angle measurements. The contact angle was primarily dictated by the outermost layer. Of the coatings studied, gelatin was the most hydrophilic. A film of (PSS/PEI)4/(gelatin/PDL)4/ gelatin was highly favorable for cell adhesion and growth, in contrast to films of (PSS/PEI)8 or (PSS/PEI)8/PSS. Cell growth patterns were successfully controlled by selective deposition of microspheres on silicone rubber, using microcontact printing with a silicone stamp. Cell adhesion was confined to the region of microsphere deposition. These results demonstrate that the LbL self-assembly technique provides a general approach to coat and selectively deposit films with nanometer thickness on silicone rubber. Furthermore, they show that this method is a viable technique for controlling cellular adhesion and growth.

Nano-encapsulation of furosemide microcrystals for controlled drug release.

Furosemide microcrystals were encapsulated with polyions and gelatin to control the release of the drug in aqueous solutions. Charged linear polyions and gelatin were alternatively deposited on 5-microm drug microcrystals through layer-by-layer (LbL) assembly. Sequential layers of poly(dimethyldiallyl ammonium chloride) (PDDA) and poly(styrenesulfonate) (PSS) were followed by adsorption of two to six gelatin/PSS bilayers with corresponding capsule wall thicknesses ranging from 45 to 115 nm. The release of furosemide from the coated microparticles was measured in aqueous solutions of pH 1.4 and 7.4. At both pH values, the release rate of furosemide from the encapsulated particles was reduced by 50-300 times (for capsules coated with two to six bilayers) compared to uncoated furosemide. The results provide a method of achieving prolonged drug release through self-assembly of polymeric shells on drug microcrystals.

Reduction of coherent scattering noise with multiple receiver Doppler.

Doppler ultrasound (US) velocity estimates are inherently subject to error as a result of both Doppler ambiguity and coherent scattering. The coherent scattering error is a result of changes in the phase of the returned echo as particles enter and leave the sample volume. This phase depends on the distance from the transmitter to the scatterer and then to the receiver. This distance, in turn, depends on the angle of the receiver. A numerical simulation has been used to determine whether velocity estimates obtained from receiver probes at different angles are independent of one another. If so, then it is possible to obtain an improved velocity estimate from the combination of several receivers at different angles. The simulation results show that the cross-correlation between velocity estimates is reduced to 0.3 when receiver probes are oriented 5 degrees apart. These results suggest a new Doppler method that can significantly reduce velocity estimation error.

Electrostatic layer-by-layer nanoassembly on biological microtemplates: platelets.

Platelets were coated with 78-nm silica nanoparticles, 45-nm fluorescent nanospheres, or bovine immunoglobulin G (IgG) through layer-by-layer assembly by alternate adsorption with oppositely charged linear polyions. Sequential deposition on platelet surfaces of cationic poly(dimethyldiallylammonium chloride) and anionic poly(styrene sulfonate) was followed by adsorption of nanoparticles or immunoglobulins. Nano-organized shells of platelets were demonstrated by transmission electron microscopy and fluorescence microscope images. Bovine IgG was assembled on platelets, as verified with anti-bovine IgG-FITC labeling. Localized targeting of anti-IgG shelled platelets was also demonstrated. An ability to coat blood cells with nano-organized shells can have applications in cardiovascular research and targeted drug delivery.