Intravascular molecular-structural imaging with a miniaturized integrated near-infrared fluorescence and ultrasound catheter

Coronary artery disease (CAD) remains a leading cause of mortality and warrants new imaging approaches to better guide clinical care. We report on a miniaturized, hybrid intravascular catheter and imaging system for comprehensive coronary artery imaging in vivo. Our catheter exhibits a total diameter of 1.0 mm (3.0 French), equivalent to standalone clinical intravascular ultrasound (IVUS) catheters but enables simultaneous near-infrared fluorescence (NIRF) and IVUS molecular-structural imaging. We demonstrate NIRF-IVUS imaging in vitro in coronary stents using NIR fluorophores, and compare NIRF signal strengths for prism and ball lens sensor designs in both low and high scattering media. Next, in vivo intravascular imaging in pig coronary arteries demonstrates simultaneous, co-registered molecular-structural imaging of experimental CAD inflammation on IVUS and distance-corrected NIRF images. The obtained results suggest substantial potential for the NIRF-IVUS catheter to advance standalone IVUS, and enable comprehensive phenotyping of vascular disease to better assess and treat patients with CAD.     

小口径血液导流管的实验研究

目的:探讨小口径血液导流管在动物离断肢体模型中快速恢复通血的实验基础应用,研究小口径血液导流管实验幼猪离断肢体维持通血效果的评价。方法:20只实验幼猪随机分为A、B两组,制成后肢完全离断模型模型,采用内径为2.0 mm、外径2.5mm的血液导流管,A组长度10 cm;B组长度20 cm,进行血管桥接后定期观察血液导流管通畅性,观察终点为血液导流管完全堵塞,血管超声探测仪无血流信号,远端血管搏动消失,离断肢体以远皮下毛细血管网无渗血。比较两组到达观察终点的时间有无差异。结果:建立临时血管通路后,离断肢体远端股动脉的远端有搏动,血管超声探测仪可检测到血液导流管内有血流信号,随着时间的延长,血液导流管动脉段逐渐由鲜红色变为暗红色,导流管段逐渐形成附壁血栓,远端血管搏动及皮下毛细管网渗血逐渐减弱直至消失,血流信号消失,两组到达观测终点的时间分别为A组365±47.4 min;B组359±31.5 min,两者比较其差异没有统计学意义(P0.05)。说明长度在10 cm-20 cm的小口径血液导流管在实验动物离断肢体血管通血方面无明显差异。结论:小口径血液导流管能够用于动物离断肢体的血管临时桥接,维持通血时间可达6-8小时,有效通血时间长。实验数据说明小口径血液导流管适合于动物离断肢体模型中的血管桥接,在下一步临床应用中在四肢复杂血管损伤中有着较为广阔的临床应用前景。     

A simple highly sensitive recording microspectrophotometer

A design of the recording microspectrophotometer is described. The instrument possesses an absolutely flat base line and quantum-noise limited detection threshold. Two principal elements of the design are the "jumping" stage, and the logarithmic amplifier with the phase-sensitive detector which converts the photomultiplier output into the optical density signal. The performance of the instrument is illustrated by the recordings of visual pigment spectra in single photoreceptors.     

Two-dimensional, computer-controlled film scanner: quantitation of fluorescence from ethidium bromide-stained DNA gels

A two-dimensional scanner based on a digital plotter is described. The device is used to analyze photographic negatives of ethidium bromide-stained DNA-agarose gels. Scanning is controlled by and photometric data transferred to a computer for processing, storage, display, and analysis such as integration of the areas under bands and determination of the mean distances of migration of polydisperse samples. An integral light source and detector module designed for reading optical "bar-codes" is mounted in place of the pen of the plotter. Spatial resolution and reproducibility are about 0.2 and 0.005 mm, respectively. Photometric precision as good as one part per thousand is achieved by sinusoidal modulation of the intensity of the light source and synchronous, phase-sensitive detection of the signal from the detector by a lock-in amplifier. No part of the sensor assembly touches the surface of the negative. In contrast to a densitometer, the computer transforms photometric data to values directly proportional to the amount of DNA at given points on the original gel. The ability to move the sensor in two dimensions over the negative allows for the integration across the width of a lane correctly allowing for the nonuniform distribution of the DNA.     

Enhanced myogenic tone in cerebral arteries from a rabbit model of subarachnoid hemorrhage

Cerebral artery vasospasm is a major cause of death and disability in patients experiencing subarachnoid hemorrhage (SAH). Currently, little is known regarding the impact of SAH on small diameter (100-200 microm) cerebral arteries, which play an important role in the autoregulation of cerebral blood flow. With the use of a rabbit SAH model and in vitro video microscopy, cerebral artery diameter was measured in response to elevations in intravascular pressure. Cerebral arteries from SAH animals constricted more (approximately twofold) to pressure within the physiological range of 60-100 mmHg compared with control or sham-operated animals. Pressure-induced constriction (myogenic tone) was also enhanced in arteries from control animals organ cultured in the presence of oxyhemoglobin, an effect independent of the vascular endothelium or nitric oxide synthesis. Finally, arteries from both control and SAH animals dilated as intravascular pressure was elevated above 140 mmHg. This study provides evidence for a role of oxyhemoglobin in impaired autoregulation (i.e., enhanced myogenic tone) in small diameter cerebral arteries during SAH. Furthermore, therapeutic strategies that improve clinical outcome in SAH patients (e.g., supraphysiological intravascular pressure) are effective in dilating small diameter cerebral arteries isolated from SAH animals.     

A phase-shift fluorometer using a laser and a transverse electrooptic modulator for subnanosecond lifetime measurements.

We described a simple phase-shift fluorometer using continuous laser excitation. The laser enables the use of a transverse mode electrooptic modulator with a half-wave retardation voltage of about 200 V (in contrast to many kilovolts of longitudinal modulators) at frequencies up to 100 MHz. The modulated fluorescence signal is detected, after passing through a double monochromator, by a photomultiplier tube feeding a radio frequency (RF) tuned amplifier. THE RF phase is then determined by phase-sensitive detection using a double balanced mixer with the reference obtained from a PIN photodiode-turned amplifier combination which detects light split off from the main exciting beam. The laser and double monochromator allow the observation of modulated Raman solvent and Rayleigh scatterin, which are convenient for determining the zero reference phase.     

Quantitative radionuclide assessment of total pulmonary vascular volume changes

Current techniques do not permit continuous and noninvasive assessments of changes in total pulmonary intravascular volume. Hence, the present study was undertaken to determine whether quantitative radionuclide imaging can be used to determine the direction and estimate the magnitude of total pulmonary vascular volume changes. The pulmonary circulation was separately perfused at a constant rate via the pulmonary artery and drained at a constant pressure via the left atrium in nine dogs. Changes in pulmonary intravascular volume were recorded as reciprocal changes in extracorporeal reservoir volume during phenylephrine or isoproterenol administration, a 20% increase in pulmonary artery flow or a 5 mmHg (1 mmHg = 133.32 Pa) decrease in left atrial pressure. Erythrocytes were labeled with technetium-99m and pulmonary volume changes were determined from tissue attenuation, blood radioactivity, and changes in total pulmonary radioactivity obtained with a gamma-camera. During each of the interventions, count changes correlated with volume changes (r greater than or equal to 0.75). The technique reliably detected volume changes as small as 10 mL. For all 531 individual pairs of radionuclide- and reservoir-determined volume changes, the correlation between reservoir-determined and radionuclide-estimated pulmonary intravascular volume changes was 0.87. The standard error of the radionuclide estimate was 21 mL. Hence, the present study demonstrates that quantitative radionuclide imaging can be used to continuously and noninvasively determine total pulmonary vascular volume changes.     

Initial hydrodynamic study on a new intraaortic axial flow pump: Dynamic aortic valve

Rotary blood pumps have been researched as implantable ventricular assist devices for years. To further reduce the complex of implanted axial pumps, the authors proposed a new concept of intraaortic axial pump, termed previously as "dynamic aortic valve (DAV)". Instead of being driven by an intraaortic micro-electric motor, it was powered by a magnetic field from outside of body. To ensure the perfusion of coronary artery, the axial flow pump is to be implanted in the position of aortic valve. It could serve as either a blood pump or a mechanical valve depending on the power input. This research tested the feasibility of the new concept in model study. A column, made from permanent magnet, is jointed to an impeller in a concentric way to form a "rotor-impeller". Supported by a hanging shaft cantilevered in the center of a rigid cage, the rotor-impeller can be turned by the magnetic field in the surrounding space. In the present prototype, the rotor is 8 mm in diameter and 15 mm in length, the impeller ha     

Intravascular pressure and diameter profile of the utero-ovarian resistance artery network: estrous cycle-dependent modulation of resistance artery tone

Blood flow to the ovary varies dramatically in both magnitude and distribution throughout the estrous cycle to meet the hormonal and metabolic demands of the ovarian parenchyma as it cyclically develops and regresses. Several vascular components appear to be critical to vascular regulation of the ovary. As a first step in resolving the role of the resistance arteries and their paired veins in regulating ovarian blood flow and transvascular exchange, we characterized the architecture and intravascular pressure profile of the utero-ovarian resistance artery network in an in vivo preparation of the ovary of the anesthetized Golden hamster. We also investigated estrous cycle-dependent changes in resistance artery tone. The right ovary and the cranial aspect of the uterus in 26 female hamsters were exposed for microcirculatory observations. Estrous-cycle phase was determined in each animal before experimentation. The utero-ovarian vascular architecture was determined and resistance artery diameters were measured in each animal by video microscopy. Servo-null intravascular pressure measurements were made throughout the uteroovarian arterial network in 11 of the animals. Architectural data showed a complex anastomotic network jointly supplying the uterus and ovary. Resistance arteries showed a high degree of coiling and close apposition to veins, maximizing countercurrent-exchange capabilities. Arterial pressure dropped below 60% of systemic arterial pressure before the arteries entered the ovary. Both the ovarian artery and the uterine artery, which jointly feed the ovary, showed cycle day-dependent changes in diameter. Arterial diameters were smallest on the day following ovulation, during the brief luteal phase of the hamster. The data show that resistance arteries comprise a critical part of a complex network designed for intimate local communication and control and suggest that these arteries may play an important role in regulating ovarian blood flow in an estrous cycle-specific manner.     

Does vibration cause poststenotic dilatation in vivo and influence atherogenesis in cholesterol-fed rabbits?

Arterial post-stenotic dilatation (PSD) is a fusiform swelling immediately down-stream to a stenosis. It is characterized by the presence of turbulent blood flow and wall vibration which has been claimed by others to be causal by producing structural weakening. We tested the hypothesis that vibration causes PSD in vivo by attaching electromagnetic and pneumatic vibrators to the aortic wall in chronic rabbits. We also observed whether mechanical vibration of the aorta in vivo influenced the distribution of oil-red-O lesions during one percent dietary cholesterol feeding. Low mass vibration gauges were developed to measure the vibration. Electromechanical vibrators having a ceramic magnet slug within a coil supplied with 50 Hz were glued to the aorta of chronic rabbits and the vibration maintained for an average of 8 weeks. Despite greater amounts of energy imparted to the wall there was no dilatation or difference in oil-red-O staining from the controls. Five weeks vibration at 100 Hz and an amplitude equal to the normal diameter pulse also produced no dilatation. We conclude that vibration does not cause PSD in vivo and suggest that its cause is likely to involve the vascular muscle stimulated by the effect of turbulent flow on the endothelium.     

A computational study on the influence of catheter-delivered intravascular probes on blood flow in a coronary artery model

Catheter-delivered intravascular probes are widely used in clinical practice to measure coronary arterial velocity and pressure, but the artefactual effect of the probe on the variables being measured is not well characterised. A coronary artery was simulated with a 180 degrees curved tube 3mm in diameter and the effect of catheters of different diameters was modelled numerically under pulsatile flow conditions. The presence of a catheter increased pressure by 1.3-4.3 mmHg depending on its diameter, and reduced velocity-pressure phase-lag. For an ultrasound sample volume 5mm downstream from the probe tip, the underestimation in velocity measurement attributed to catheter blockage is approximately 15-21% for an average inlet velocity of 0.1m/s. The velocity measurement error is lower at higher mean flow velocity. Accuracy of clinical velocity measurements could be improved by moving the sample volume farther downstream from the probe tip, because the centrifugal pressure gradient intrinsic to the curvature promotes re-development of flow.     

A general amphipathic alpha-helical motif for sensing membrane curvature

The Golgi-associated protein ArfGAP1 has an unusual membrane-adsorbing amphipathic alpha-helix: its polar face is weakly charged, containing mainly serine and threonine residues. We show that this feature explains the specificity of ArfGAP1 for curved versus flat lipid membranes. We built an algorithm to identify other potential amphipathic alpha-helices rich in serine and threonine residues in protein databases. Among the identified sequences, we show that three act as membrane curvature sensors. In the golgin GMAP-210, the sensor may serve to trap small vesicles at the end of a long coiled coil. In Osh4p/Kes1p, which transports sterol between membranes, the sensor controls access to the sterol-binding pocket. In the nucleoporin Nup133, the sensor corresponds to an exposed loop of a beta-propeller structure. Ser/Thr-rich amphipathic helices thus define a general motif used by proteins of various functions for sensing membrane curvature.     

Exercise hyperemia and vasoconstrictor responses in humans with cystic fibrosis.

ATP released from circulating erythrocytes is a potential signal regulating muscle blood flow during exercise (exercise hyperemia), and intravascular ATP appears to blunt sympathetic vasoconstriction during exercise. Erythrocytes from patients with cystic fibrosis (CF) do not release ATP. The goal of the present study was to determine whether increases in forearm blood flow during exercise are blunted in CF patients and whether CF patients exhibit greater vasoconstrictor responsiveness during exercise. Nine control subjects and 10 CF patients who were free of other disease complications (approximately 96% O2 saturation) performed incremental rhythmic forearm exercise at 5, 10, and 15% of maximum handgrip strength for 21 min (7 min at each workload). We used a cold pressor test to evoke sympathetic vasoconstriction under resting conditions and at each exercise workload. As a control, subjects performed a second exercise bout without the cold pressor test. Continuous brachial artery blood velocity was monitored beat-to-beat, and vessel diameter was assessed by Doppler ultrasound. Artery diameter, as well as blood pressure, heart rate, and O2 saturation, was measured at steady-state exercise and at 1 min into the cold pressor stimulus. Blood pressure and heart rate responses to the forearm exercise and each cold pressor test were similar in both groups (P > 0.05). Contrary to our hypothesis, forearm blood flow (P = 0.91) and forearm vascular conductance (P = 0.82) were similar at rest and at each level of exercise between CF patients and controls. Additionally, there was no difference in the degree of sympathetic vasoconstriction between groups at rest and at each level of exercise (P = 0.22). Our results suggest that ATP released from the deformation of erythrocytes is not an obligatory signal for exercise hyperemia in human skeletal muscle.     

体外循环血栓非侵入在线电阻抗成像方法

目的 在体外循环系统中,血栓的在线检测和可视化具有重要意义。本文提出了基于电阻抗成像（EIT）的体外循环血栓非侵入在线检测方法。方法 首先通过联合仿真研究了传感器尺寸对成像效果的影响。其次,根据仿真结果设计了直径为20 mm的16铜质电极EIT传感器,搭建了循环流动实验平台,并设计了静态和循环流动实验。使用尺寸为3～6 mm的猪血块代替血栓,将血块置于新鲜猪血样本中,采用Tikhonov正则化算法进行成像。将3 mm和5 mm的血块分别置于循环系统中,重建血块在传感器截面的大小和位置图像,并与高速相机拍摄结果进行对比。结果 仿真结果显示当目标物与传感器面积比（AR）不小于0.01时,传感器直径为20 mm和30 mm对应的图像相关系数（IC）均大于0.06,成像效果较好。静态成像结果显示,相对尺寸覆盖率误差（RCR）小于等于0.1。循环流动实验显示,血块经过传感器时,检测到归一化后的相对电导率变化值分别为80和200,结果显示该方法能够检测到循环系统中的血块。结论 该方法具有实时性和非侵入的优点,有望应用于体外血栓的检测。     

High speed solution of 2nd order curves with special application to planar sections of blood vessels

Rapid determination of the areas of planar sections through a blood vessel are required in an on-line blood flow measurement system which utilizes an intravascular ultrasonic catheter. Vessel areas are determined by: (1) using a fast algorithm to fit an ellipse to 6 data points of the vessel wall's location measured with the ultrasonic catheter; (2) computing the area of the determined ellipse. The program is implemented in a general purpose computer with a hardware floating point processor, using 3320 of 16 bit words of memory for storing variables, data, look-up tables, and computation instructions. The program executes in 3 ms, which, when combined with the other computations and timing requirements of the system, allows instantaneous flow computation every 7.3 ms. The system is fast enough to follow pulsitile blood flow. The algorithm applies in other areas which also require fast quantitative measurement, for example, in determining organ or tissue structural contours describable with a 2nd order equation.     

Initial hydrodynamic study on a new intraaortic axial flow pump: Dynamic aortic valve

Rotary blood pumps have been researched as implantable ventricular assist devices for years. To further reduce the complex of implanted axial pumps, the authors proposed a new concept of intraaortic axial pump, termed previously as “dynamic aortic valve (DAV)”. Instead of being driven by an intraaortic micro-electric motor, it was powered by a magnetic field from outside of body. To ensure the perfusion of coronary artery, the axial flow pump is to be implanted in the position of aortic valve. It could serve as either a blood pump or a mechanical valve depending on the power input. This research tested the feasibility of the new concept in model study. A column, made from permanent magnet, is jointed to an impeller in a concentric way to form a “rotor-impeller”. Supported by a hanging shaft cantilevered in the center of a rigid cage, the rotor-impeller can be turned by the magnetic field in the surrounding space. In the present prototype, the rotor is 8 mm in diameter and 15 mm in length, the impeller has 3 vanes with an outer diameter of 18 mm. The supporting cage is 22 mm in outer diameter and 20 mm in length. When tested, the DAV prototype is inserted into the tube of a mock circuit. The alternative magnetic field is produced by a rotating magnet placed side by side with the rotor-impeller at a distance of 30 mm. Once the alternative magnetic field is presented in the surrounding space, the DAV starts to turn, leading to a pressure difference and liquid flow in the tube. The flow rate or pressure difference is proportioned to rotary speed. At the maximal output of hydraulic power, the flow rate reached 5 L/min against an afterload of 100 mmHg. The maximal pressure difference generated by DAV at a rotation rate of 12600 r/min was 147 mmHg. The preliminary results demonstrated the feasibility of “DAV”, further research on this concept is justifiable.     

Pulsatile flow in a catheterised artery

A model is presented for the physiological problem of a catheter which is inserted in a femoral artery to measure the pressure gradient. As the catheter will modify the pressure distribution in the artery, the pressure gradient which would be recorded by a perfect pressure transducer attached to it would differ from that in the uncatheterised artery. To estimate the magnitude of this error, it is assumed that the rates of flow of blood through the catheterised and the uncatheterised artery are described by the same known periodic function of time.     

Coupled modeling of blood perfusion in intravascular, interstitial spaces in tumor microvasculature

The coupling of intravascular and interstitial flow is a distinct feature of tumor microcirculation, due to the high vessel permeability, the low osmotic pressure gradient as well as the absence of functional lymphatic system inside tumors. In this paper, a coupled mathematical model of tumor microcirculation is developed, which provides the link between microvasculature and interstitial space perfusion through the matrices determining a neighbor point belonging to either connected vessel (matrix B) or interstitial space (matrix A), and combines the intravascular and interstitial flow by vascular leaky terms. In addition, the compliance of tumor vessels, blood rheology with hematocritic distribution at branches is also considered. The microvascular network, on which the microcirculation calculation is carried out, is generated from our two-dimensional 9-point (2D9P) model of tumor angiogenesis, improved from the previous 2D5P one. A specific coupling procedure is developed in the study to couple the intravascular and interstitial flow. It is based on the iteratively numerical simulation techniques, including local iterations at individual parameter level and one global loop to provide coupling and simulation convergence. The simulation results not only present the basic features and characteristics of tumor microcirculation, which agree with the corresponding experimental observations reported, but also predict an intimate relationship between the tumor intravascular and interstitial flow quantitatively. Among the parameters, the vascular leakiness is a key to govern the systemic flowing pattern, influence the tumor internal environment and contribute to the metastasis of tumor cells, which could not be presented by the previous uncoupled models.     

Functional implications of the human T-lymphotropic virus type 1 transmembrane glycoprotein helical hairpin structure

Retrovirus entry into cells follows receptor binding by the surface-exposed envelope glycoprotein (Env) subunit (SU), which triggers the membrane fusion activity of the transmembrane (TM) protein. TM protein fragments expressed in the absence of SU adopt helical hairpin structures comprising a central coiled coil, a region of chain reversal containing a disulfide-bonded loop, and a C-terminal segment that packs onto the exterior of the coiled coil in an antiparallel manner. Here we used in vitro mutagenesis to test the functional role of structural elements observed in a model helical hairpin, gp21 of human T-lymphotropic virus type 1. Membrane fusion activity requires the stabilization of the N and C termini of the central coiled coil by a hydrophobic N cap and a small hydrophobic core, respectively. A conserved Gly-Gly hinge motif preceding the disulfide-bonded loop, a salt bridge that stabilizes the chain reversal region, and interactions between the C-terminal segment and the coiled coil are also critical for fusion activity. Our data support a model whereby the chain reversal region transmits a conformational signal from receptor-bound SU to induce the fusion-activated helical hairpin conformation of the TM protein.     

The Influence of Drag on Nonlinear Oscillatory Flow through Concentric Annulus

A mathematical model has been developed to study the effect of particle drag parameter and frequency parameter on velocity and pressure gradient in nonlinear oscillatory two phase flow. The main purpose is to apply the model to study the combined effect of introduction of the catheter and elastic properties of the arterial wall on the pulsatile nature of the blood flow. We model the artery as an isotropic thin walled elastic tube and the catheter as a coaxial flexible tube. Blood is modeled as an incompressible particulate viscous Newtonian fluid. Perturbation technique has been applied to find the approximations for velocity and pressure gradient up to second order. Numerical solutions are investigated with graphical presentations to understand the effects of drag parameter, frequency parameter and phase angle on velocity along radial direction and pressure gradient along axial directions. As the drag parameter increases, mean pressure gradient and mean velocity will be decreased. As frequency parameter increases mean velocity profile bends near the outer wall. Due to elastic nature of artery wall, a thin catheter experience small oscillations and a thick catheter remains stationary inside the artery. Finally, the effect of catheterization on various physiologically important flow rate characteristics—mean velocity, mean pressure gradient are studied for a range of different catheter sizes, particle drag parameter and frequency parameters.