Integrated method for quantitative morphometry and oxygen transport modeling in striated muscle.

Identifying structural limitations in O2 transport is primarily restricted by current methods employed to characterize the nature of physiological remodeling. Inadequate resolution or breadth of available data has impaired development of routine diagnostic protocols and effective therapeutic strategies. Understanding O2 transport within striated muscle faces major challenges, most notably in quantifying how well individual fibers are supplied by the microcirculation, which has necessitated exploring tissue O2 supply using theoretical modeling of diffusive exchange. With capillary domains identified as a suitable model for the description of local O2 supply and requiring less computation than numerically calculating the trapping regions that are supplied by each capillary via biophysical transport models, we sought to design a high-throughput method for histological analysis. We present an integrated package that identifies optimal protocols for identification of important input elements, processing of digitized images with semiautomated routines, and incorporation of these data into a mathematical modeling framework with computed output visualized as the tissue partial pressure of O2 (Po2) distribution across a biopsy sample. Worked examples are provided using muscle samples from experiments involving rats and humans. NEW & NOTEWORTHY Progress in quantitative morphometry and analytical modeling has tended to develop independently. Real diagnostic power lies in harnessing both disciplines within one user-friendly package. We present a semiautomated, high-throughput tool for determining muscle phenotype from biopsy material, which also provides anatomically relevant input to quantify tissue oxygenation, in a coherent package not previously available to nonspecialist investigators.

Real-Time 2-D Phased Array Vector Flow Imaging.

Echocardiography examination of the blood flow is currently either restricted to 1-D techniques in real-time or experimental offline 2-D methods. This paper presents an implementation of transverse oscillation for real-time 2-D vector flow imaging (VFI) on a commercial BK Ultrasound scanner. A large field-of-view (FOV) sequence for studying flow dynamics at 11 frames per second (fps) and a sequence for studying peak systolic velocities (PSVs) with a narrow FOV at 36 fps were validated. The VFI sequences were validated in a flow rig with continuous laminar parabolic flow and in a pulsating flow pump system before being tested in vivo, where measurements were obtained on two healthy volunteers. Mean PSV from 11 cycles was 155 cms-1 with a precision of ±9.0% for the pulsating flow pump. In vivo, PSV estimated in the ascending aorta was 135 cms-1 ± 16.9% for eight cardiac cycles. Furthermore, in vivo flow dynamics of the left ventricle and in the ascending aorta were visualized. In conclusion, angle independent 2-D VFI on a phased array has been implemented in real time, and it is capable of providing quantitative and qualitative flow evaluations of both the complex and fully transverse flow.

A Comparison Study of Vector Velocity, Spectral Doppler and Magnetic Resonance of Blood Flow in the Common Carotid Artery.

Magnetic resonance phase contrast angiography (MRA) is the gold standard for blood flow evaluation. Spectral Doppler ultrasound (SDU) is the first clinical choice, although the method is angle dependent. Vector flow imaging (VFI) is an angle-independent ultrasound method. The aim of the study was to compare VFI- and SDU-estimated peak systolic velocities (PSV) of the common carotid artery (CCA) with PSV obtained by MRA. Furthermore, intra- and inter-observer agreement was determined. MRA estimates were significantly different from SDU estimates (left CCA: p < 0.001, right CCA: p < 0.001), but not from VFI estimates (left CCA: p = 0.28, right CCA: p = 0.18). VFI measured lower PSV in both CCAs compared with SDU (p < 0.001) with improved precision (VFI: left: 24%, right: 18%; SDU: left 38%, right: 23%). Intra- and inter-observer agreement was almost perfect for VFI and SDU (inter-observer correlation coefficient: VFI 0.88, SDU 0.91; intra-observer correlation coefficient: VFI 0.96, SDU 0.97). VFI is more accurate than SDU in evaluating PSV compared with MRA.

Accuracy and Precision of a Plane Wave Vector Flow Imaging Method in the Healthy Carotid Artery.

The objective of the study described here was to investigate the accuracy and precision of a plane wave 2-D vector flow imaging (VFI) method in laminar and complex blood flow conditions in the healthy carotid artery. The approach was to study (i) the accuracy for complex flow by comparing the velocity field from a computational fluid dynamics (CFD) simulation to VFI estimates obtained from the scan of an anthropomorphic flow phantom and from an in vivo scan; (ii) the accuracy for laminar unidirectional flow in vivo by comparing peak systolic velocities from VFI with magnetic resonance angiography (MRA); (iii) the precision of VFI estimation in vivo at several evaluation points in the vessels. The carotid artery at the bifurcation was scanned using both fast plane wave ultrasound and MRA in 10 healthy volunteers. The MRA geometry acquired from one of the volunteers was used to fabricate an anthropomorphic flow phantom, which was also scanned using the fast plane wave sequence. The same geometry was used in a CFD simulation to calculate the velocity field. Results indicated that similar flow patterns and vortices were estimated with CFD and VFI in the phantom for the carotid bifurcation. The root-mean-square difference between CFD and VFI was within 0.12 m/s for velocity estimates in the common carotid artery and the internal branch. The root-mean-square difference was 0.17 m/s in the external branch. For the 10 volunteers, the mean difference between VFI and MRA was -0.17 m/s for peak systolic velocities of laminar flow in vivo. The precision in vivo was calculated as the mean standard deviation (SD) of estimates aligned to the heart cycle and was highest in the center of the common carotid artery (SD = 3.6% for velocity magnitudes and 4.5° for angles) and lowest in the external branch and for vortices (SD = 10.2% for velocity magnitudes and 39° for angles). The results indicate that plane wave VFI measures flow precisely and that estimates are in good agreement with a CFD simulation and MRA.

Vector and Doppler Ultrasound Velocities Evaluated in a Flow Phantom and the Femoropopliteal Vein.

Ultrasound is used for evaluating the veins of the lower extremities. Operator and angle dependency limit spectral Doppler ultrasound (SDUS). The aim of the study was to compare peak velocity measurements in a flow phantom and the femoropopliteal vein of 20 volunteers with the angle-independent vector velocity technique vector flow imaging (VFI) and SDUS. In the flow phantom, VFI underestimated velocity (p = 0.01), with a lower accuracy of 5.5% (p = 0.01) and with no difference in precision, that is, error factor, compared with SDUS (VFI: 1.02 vs. SDUS: 1.02, p = 0.58). In vivo, VFI estimated lower velocities (femoral: p = 0.001; popliteal: p = 0.001) with no difference in precision compared with SDUS (femoral: VFI 1.09 vs. SDUS 1.14, p = 0.37; popliteal: VFI 1.13 vs. SDUS 1.06, p = 0.09). In conclusion, the precise VFI technique can be used to characterize venous hemodynamics of the lower extremities despite its underestimation of velocities.

Common Carotid Artery Flow Measured by 3-D Ultrasonic Vector Flow Imaging and Validated with Magnetic Resonance Imaging.

Ultrasound (US) examination of the common carotid artery was compared with a through-plane magnetic resonance imaging (MRI) sequence to validate a recently proposed technique for 3-D US vector flow imaging. Data from the first volunteer examined were used as the training set, before volume flow and peak velocities were calculated for the remaining eight volunteers. Peak systolic velocities (PSVs) and volume flow obtained with 3-D US were, on average, 34% higher and 24% lower than those obtained with MRI, respectively. A high correlation was observed for PSV (r = 0.79), whereas a lower correlation was observed for volume flow (r = 0.43). The overall standard deviations were ±5.7% and ±5.7% for volume flow and PSV with 3-D US, compared with ±2.7% and ±3.2% for MRI. Finally, the data were re-processed with a change in the parameter settings for the echo-canceling filter to investigate its influence on overall performance. PSV was less affected by the re-processing, whereas the difference in volume flow between 3-D vector flow imaging and MRI was reduced to -9%, and with an improved overall standard deviation of ±4.7%. The results illustrate the feasibility of using 3-D US for precise and angle-independent volume flow and PSV estimation in vivo.

Ultrasonic 3-D Vector Flow Method for Quantitative In Vivo Peak Velocity and Flow Rate Estimation.

Current clinical ultrasound (US) systems are limited to show blood flow movement in either 1-D or 2-D. In this paper, a method for estimating 3-D vector velocities in a plane using the transverse oscillation method, a 32×32 element matrix array, and the experimental US scanner SARUS is presented. The aim of this paper is to estimate precise flow rates and peak velocities derived from 3-D vector flow estimates. The emission sequence provides 3-D vector flow estimates at up to 1.145 frames/s in a plane, and was used to estimate 3-D vector flow in a cross-sectional image plane. The method is validated in two phantom studies, where flow rates are measured in a flow-rig, providing a constant parabolic flow, and in a straight-vessel phantom ( ∅=8 mm) connected to a flow pump capable of generating time varying waveforms. Flow rates are estimated to be 82.1 ± 2.8 L/min in the flow-rig compared with the expected 79.8 L/min, and to 2.68 ± 0.04 mL/stroke in the pulsating environment compared with the expected 2.57 ± 0.08 mL/stroke. Flow rates estimated in the common carotid artery of a healthy volunteer are compared with magnetic resonance imaging (MRI) measured flow rates using a 1-D through-plane velocity sequence. Mean flow rates were 333 ± 31 mL/min for the presented method and 346 ± 2 mL/min for the MRI measurements.

3-D Vector Flow Estimation With Row-Column-Addressed Arrays.

Simulation and experimental results from 3-D vector flow estimations for a 62 + 62 2-D row-column (RC) array with integrated apodization are presented. A method for implementing a 3-D transverse oscillation (TO) velocity estimator on a 3-MHz RC array is developed and validated. First, a parametric simulation study is conducted, where flow direction, ensemble length, number of pulse cycles, steering angles, transmit/receive apodization, and TO apodization profiles and spacing are varied, to find the optimal parameter configuration. The performance of the estimator is evaluated with respect to relative mean bias ~B and mean standard deviation ~σ . Second, the optimal parameter configuration is implemented on the prototype RC probe connected to the experimental ultrasound scanner SARUS. Results from measurements conducted in a flow-rig system containing a constant laminar flow and a straight-vessel phantom with a pulsating flow are presented. Both an M-mode and a steered transmit sequence are applied. The 3-D vector flow is estimated in the flow rig for four representative flow directions. In the setup with 90° beam-to-flow angle, the relative mean bias across the entire velocity profile is (-4.7, -0.9, 0.4)% with a relative standard deviation of (8.7, 5.1, 0.8)% for ( vx, vy, vz ). The estimated peak velocity is 48.5 ± 3 cm/s giving a -3% bias. The out-of-plane velocity component perpendicular to the cross section is used to estimate volumetric flow rates in the flow rig at a 90° beam-to-flow angle. The estimated mean flow rate in this setup is 91.2 ± 3.1 L/h corresponding to a bias of -11.1%. In a pulsating flow setup, flow rate measured during five cycles is 2.3 ± 0.1 mL/stroke giving a negative 9.7% bias. It is concluded that accurate 3-D vector flow estimation can be obtained using a 2-D RC-addressed array.

Adaptations to Speed Endurance Training in Highly Trained Soccer Players.

PURPOSE: The present study examined whether a period of additional speed endurance training would improve intense intermittent exercise performance in highly trained soccer players during the season and whether the training changed aerobic metabolism and the level of oxidative enzymes in type I and type II muscle fibers. METHODS: During the last 9 wk of the season, 13 semiprofessional soccer players performed additional speed endurance training sessions consisting of two to three sets of 8-10 repetitions of 30-m sprints with 10 s of passive recovery (SET). Before and after SET, subjects completed a double-step exercise protocol that included transitions from standing to moderate-intensity running (~75% HRmax), followed by transitions from moderate- to high-intensity running (~90% HRmax) in which pulmonary oxygen uptake (V˙O2) was determined. In addition, the yo-yo intermittent recovery test level 1 was performed, and a muscle biopsy was obtained at rest. RESULTS: The yo-yo intermittent recovery test level 1 performance was 11.6% ± 6.4% (mean ± SD) better (2803 ± 330 vs 3127 ± 383 m, P < 0.05) after SET compared with before SET. In the transition from standing to moderate-intensity running, phase II pulmonary V˙O2 kinetics was 11.4% ± 16.5% faster (P < 0.05), and the running economy at this intensity was 2.3% ± 3.0% better (P < 0.05). These improvements were apparent despite the content of muscle proteins regulating oxidative metabolism (3-hydroxyacyl CoA dehydrogenase, COX IV, and OXPHOS), and capillarization was reduced (P < 0.05). The content of 3-hydroxyacyl CoA dehydrogenase and citrate synthase in type I and type II fibers did not change. CONCLUSION: In highly trained soccer players, additional speed endurance training is associated with an improved ability to perform repeated high-intensity work. To what extent the training-induced changes in V˙O2 kinetics and mechanical efficiency in type I fibers caused the improvement in performance warrants further investigation.

Safety Assessment of Advanced Imaging Sequences I: Measurements.

A method for rapid measurement of intensities (I(spta)), mechanical index (MI), and probe surface temperature for any ultrasound scanning sequence is presented. It uses the scanner's sampling capability to give an accurate measurement of the whole imaging sequence for all emissions to yield the true distributions. The method is several orders of magnitude faster than approaches using an oscilloscope, and it also facilitates validating the emitted pressure field and the scanner's emission sequence software. It has been implemented using the experimental synthetic aperture real-time ultrasound system (SARUS) scanner and the Onda AIMS III intensity measurement system (Onda Corporation, Sunnyvale, CA, USA). Four different sequences have been measured: a fixed focus emission, a duplex sequence containing B-mode and flow emissions, a vector flow sequence with B-mode and flow emissions in 17 directions, and finally a SA duplex flow sequence. A BK8820e (BK Medical, Herlev, Denmark) convex array probe is used for the first three sequences and a BK8670 linear array probe for the SA sequence. The method is shown to give the same intensity values within 0.24% of the AIMS III Soniq 5.0 (Onda Corporation, Sunnyvale, CA, USA) commercial intensity measurement program. The approach can measure and store data for a full imaging sequence in 3.8-8.2 s per spatial position. Based on I(spta), MI, and probe surface temperature, the method gives the ability to determine whether a sequence is within U.S. FDA limits, or alternatively indicate how to scale it to be within limits.

Moderate stem-cell telomere shortening rate postpones cancer onset in a stochastic model.

Mammalian cells are restricted from proliferating indefinitely. Telomeres at the end of each chromosome are shortened at cell division and when they reach a critical length, the cell will enter permanent cell cycle arrest-a state known as senescence. This mechanism is thought to be tumor suppressing, as it helps prevent precancerous cells from dividing uncontrollably. Stem cells express the enzyme telomerase, which elongates the telomeres, thereby postponing senescence. However, unlike germ cells and most types of cancer cells, stem cells only express telomerase at levels insufficient to fully maintain the length of their telomeres, leading to a slow decline in proliferation potential. It is not yet fully understood how this decline influences the risk of cancer and the longevity of the organism. We here develop a stochastic model to explore the role of telomere dynamics in relation to both senescence and cancer. The model describes the accumulation of cancerous mutations in a multicellular organism and creates a coherent theoretical framework for interpreting the results of several recent experiments on telomerase regulation. We demonstrate that the longest average cancer-free lifespan before cancer onset is obtained when stem cells start with relatively long telomeres that are shortened at a steady rate at cell division. Furthermore, the risk of cancer early in life can be reduced by having a short initial telomere length. Finally, our model suggests that evolution will favor a shorter than optimal average cancer-free lifespan in order to postpone cancer onset until late in life.