Observations of cardiac beating behaviors of wild‐type and mutant Drosophilae with optical coherence tomography

Time‐resolved optical coherence tomography (OCT) scanning images of wild‐type and mutant fruit flies (Drosophila melanogaster), illustrating the heartbeat patterns for evaluating their cardiac functions, are demonstrated. Based on the heartbeat patterns, the beat rate and the relative phase between the first two heart segments can be evaluated. The OCT scanning results of mutant flies with impaired proteasome function in cardiac muscles show irregular heartbeat patterns and systematically decreased average beat rates, when compared with the regular patterns of ～4.97 beats/s in average beat rate of the wild‐type. In both wild‐type and proteasome mutant flies, the beatings at different locations in the same heart segment are essentially synchronized. However, between different heart segments, although the beating in the second segment shows a lag in phase behind that of the first segment in a wild‐type, in a proteasome mutant, the beating in the second segment becomes significantly leading that of the first segment. Besides the comparison between the wild‐type and proteasomal mutant flies, the influences of using different methods for immobilizing flies during OCT scanning on the heart functions are demonstrated. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Speckle variance optical coherence tomography of blood flow in the beating mouse embryonic heart

Efficient separation of blood and cardiac wall in the beating embryonic heart is essential and critical for experiment‐based computational modelling and analysis of early‐stage cardiac biomechanics. Although speckle variance optical coherence tomography (SV‐OCT) relying on calculation of intensity variance over consecutively acquired frames is a powerful approach for segmentation of fluid flow from static tissue, application of this method in the beating embryonic heart remains challenging because moving structures generate SV signal indistinguishable from the blood. Here, we demonstrate a modified four‐dimensional SV‐OCT approach that effectively separates the blood flow from the dynamic heart wall in the beating mouse embryonic heart. The method takes advantage of the periodic motion of the cardiac wall and is based on calculation of the SV signal over the frames corresponding to the same phase of the heartbeat cycle. Through comparison with Doppler OCT imaging, we validate this speckle‐based approach and show advantages in its insensitiveness to the flow direction and velocity as well as reduced influence from the heart wall movement. This approach has a potential in variety of applications relying on visualization and segmentation of blood flow in periodically moving structures, such as mechanical simulation studies and finite element modelling. Picture : Four‐dimensional speckle variance OCT imaging shows the blood flow inside the beating heart of an E8.5 mouse embryo.     

Retinal pulse wave velocity measurement using spectral‐domain optical coherence tomography

The human eyes provide a natural window for noninvasive measurement of the pulse wave velocity (PWV) of small arteries. By measuring the retinal PWV, the stiffness of small arteries can be assessed, which may better detect early vascular diseases. Therefore, retinal PWV measurement has attracted increasing attention. In this study, a jump‐scanning method was proposed for noninvasive measurement of retinal PWV using spectral‐domain optical coherence tomography (SD‐OCT). The jump‐scanning method uses the phase‐resolved Doppler OCT to obtain the pulse shapes. To realize PWV measurement, the jump‐scanning method extracts the transit time of the pulse wave from an original OCT scanning site to another through a transient jump. The measured retinal arterial PWV of a young human subject with normal blood pressure was in the order of 20 to 30 mm/s, which was consistent with previous studies. As a comparison, PWV of 50 mm/s was measured for a young human subject with prehypertension, which was in accordance with the finding of strong association between retinal PWV and blood pressure. In summary, it is believed the proposed jump‐scanning method could benefit the research and diagnosis of vascular diseases through the window of human eyes.      

Joint dynamic imaging of morphogenesis and function in the developing heart

In the developing heart, time-lapse imaging is particularly challenging. Changes in heart morphology due to tissue growth or long-term reorganization are difficult to follow because they are much subtler than the rapid shape changes induced by the heartbeat. Therefore, imaging heart development usually requires slowing or stopping the heart. This, however, leads to information loss about the unperturbed heart shape and the dynamics of heart function. To overcome this limitation, we have developed a non-invasive heart imaging technique to jointly document heart function (at fixed stages of development) as well as its morphogenesis (at any fixed phase in the heartbeat) that does not require stopping or slowing the heart. We review the challenges for imaging heart development and our methodology, which is based on computationally combining and analyzing multiple high-speed image sequences acquired throughout the course of development. We present results obtained in the developing zebrafish heart. Image analysis of the acquired data yielded blood flow velocity maps and made it possible to follow the relative movement of individual cells over several hours.     

Doppler Optical Coherence Tomography of Retinal Circulation

Noncontact retinal blood flow measurements are performed with a Fourier domain optical coherence tomography (OCT) system using a circumpapillary double circular scan (CDCS) that scans around the optic nerve head at 3.40 mm and 3.75 mm diameters. The double concentric circles are performed 6 times consecutively over 2 sec. The CDCS scan is saved with Doppler shift information from which flow can be calculated. The standard clinical protocol calls for 3 CDCS scans made with the OCT beam passing through the superonasal edge of the pupil and 3 CDCS scan through the inferonal pupil. This double-angle protocol ensures that acceptable Doppler angle is obtained on each retinal branch vessel in at least 1 scan. The CDCS scan data, a 3-dimensional volumetric OCT scan of the optic disc scan, and a color photograph of the optic disc are used together to obtain retinal blood flow measurement on an eye. We have developed a blood flow measurement software called "Doppler optical coherence tomography of retinal circulation" (DOCTORC). This semi-automated software is used to measure total retinal blood flow, vessel cross section area, and average blood velocity. The flow of each vessel is calculated from the Doppler shift in the vessel cross-sectional area and the Doppler angle between the vessel and the OCT beam. Total retinal blood flow measurement is summed from the veins around the optic disc. The results obtained at our Doppler OCT reading center showed good reproducibility between graders and methods (<10%). Total retinal blood flow could be useful in the management of glaucoma, other retinal diseases, and retinal diseases. In glaucoma patients, OCT retinal blood flow measurement was highly correlated with visual field loss (R²>0.57 with visual field pattern deviation). Doppler OCT is a new method to perform rapid, noncontact, and repeatable measurement of total retinal blood flow using widely available Fourier-domain OCT instrumentation. This new technology may improve the practicality of making these measurements in clinical studies and routine clinical practice.     

Validation of a mouse conductance system to determine LV volume: comparison to echocardiography and crystals

The application of left ventricular pressure-volume analysis to transgenic mice to characterize the cardiac phenotype has been problematic due to the small size of the mouse heart and the rapid heartbeat. Conductance technology has been miniaturized for the mouse and can solve this problem. However, there has been no validation of this technique. Accordingly, we performed echocardiography followed by simultaneous ultrasonic crystals, flow probe, and conductance studies in 18 CD-1 mice. Raw conductance volumes were corrected for an inhomogenous electrical field (alpha) and parallel conductance (G(pi)) yielding a stroke volume of 14.1 +/- 3.7 microliter/beat, end-diastolic volume of 20.8 +/- 6.5 microliter, and end-systolic volume of 9.0 +/- 5.8 microliter. The mean conductance volumes were no different from those derived by flow probe and echocardiography but did differ from ultrasonic crystals. G(pi) was determined to be 14.9 +/- 8.7 microliter. However, hypertonic saline altered dimension and pressure in the mouse left ventricle. Although G(pi) can be determined by the hypertonic saline method, saline altered hemodynamics, questioning its validity in the mouse. Although mean measures of absolute volume may be similar among different techniques, individual values did not correlate.     

Doppler optical coherence tomography for measuring flow in engineered tissue

The engineering of human tissue represents a major paradigm shift in clinical medicine. Early embodiments of tissue engineering are currently being taken forward to the clinic by production methods that are essentially extensions of laboratory manual procedures. However, to achieve the status of routine large-scale clinical practice, automation and scale-out processes are required. This in turn will require the development of reliable on-line monitoring and control systems. This paper examines one demand of crucial importance, namely the real time in vitro monitoring of the flow characteristics through growing tissue since this has a complex interrelationship. Doppler optical coherence tomography (DOCT) is a recently developed imaging technique for studying the rheological properties of tissues in vivo. Capable of non-invasive imaging in real time with high resolution, it is potentially ideal for the continuous monitoring of engineered tissues in vitro. As a base line, the current status of DOCT in vivo is therefore reviewed. This paper also reports the first preliminary use of DOCT in tissue engineering. The application described involves the imaging of a fully developed laminar flow through a combined tissue fabrication/bioreactor with a tissue-engineered construct (substitute blood vessel) in situ.     

Functional morphology and patterns of blood flow in the heart of Python regius

Brightness‐modulated ultrasonography, continuous‐wave Doppler, and pulsed‐wave Doppler‐echocardiography were used to analyze the functional morphology of the undisturbed heart of ball pythons. In particular, the action of the muscular ridge and the atrio‐ventricular valves are key features to understand how patterns of blood flow emerge from structures directing blood into the various chambers of the heart. A step‐by‐step image analysis of echocardiographs shows that during ventricular diastole, the atrio‐ventricular valves block the interventricular canals so that blood from the right atrium first fills the cavum venosum, and blood from the left atrium fills the cavum arteriosum. During diastole, blood from the cavum venosum crosses the muscular ridge into the cavum pulmonale. During middle to late systole the muscular ridge closes, thus prohibiting further blood flow into the cavum pulmonale. At the same time, the atrio‐ventricular valves open the interventricular canal and allow blood from the cavum arteriosum to flow into the cavum venosum. In the late phase of ventricular systole, all blood from the cavum pulmonale is pressed into the pulmonary trunk; all blood from the cavum venosum is pressed into both aortas. Quantitative measures of blood flow volume showed that resting snakes bypass the pulmonary circulation and shunt about twice the blood volume into the systemic circulation as into the pulmonary circulation. When digesting, the oxygen demand of snakes increased tremendously. This is associated with shunting more blood into the pulmonary circulation. The results of this study allow the presentation of a detailed functional model of the python heart. They are also the basis for a functional hypothesis of how shunting is achieved. Further, it was shown that shunting is an active regulation process in response to changing demands of the organism (here, oxygen demand). Finally, the results of this study support earlier reports about a dual pressure circulation in Python regius. J. Morphol., 2009. © 2008 Wiley‐Liss, Inc.     

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient’s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.     

Joint dynamic imaging of morphogenesis and function in the developing heart

In the developing heart, time-lapse imaging is particularly challenging. Changes in heart morphology due to tissue growth or long-term reorganization are difficult to follow because they are much subtler than the rapid shape changes induced by the heartbeat. Therefore, imaging heart development usually requires slowing or stopping the heart. This, however, leads to information loss about the unperturbed heart shape and the dynamics of heart function. To overcome this limitation, we have developed a non-invasive heart imaging technique to jointly document heart function (at fixed stages of development) as well as its morphogenesis (at any fixed phase in the heartbeat) that does not require stopping or slowing the heart. We review the challenges for imaging heart development and our methodology, which is based on computationally combining and analyzing multiple high-speed image sequences acquired throughout the course of development. We present results obtained in the developing zebrafish heart. Image analysis of the acquired data yielded blood flow velocity maps and made it possible to follow the relative movement of individual cells over several hours.Key words: cardiac imaging, zebrafish, fluorescence imaging, heart development, registration, fast imaging     

Phase dual‐slopes in frequency‐domain near‐infrared spectroscopy for enhanced sensitivity to brain tissue: First applications to human subjects

We present a first in vivo application of phase dual‐slopes (DS?), measured with frequency‐domain near‐infrared spectroscopy on four healthy human subjects, to demonstrate their enhanced sensitivity to cerebral hemodynamics. During arterial blood pressure oscillations elicited at a frequency of 0.1 Hz, we compare three different ways to analyze either intensity (I) or phase (?) data collected on the subject's forehead at multiple source‐detector distances: Single‐distance, single‐slope and DS. Theoretical calculations based on diffusion theory show that the method with the deepest maximal sensitivity (at about 11 mm) is DS?. The in vivo results indicate a qualitative difference of phase data (especially DS?) and intensity data (especially single‐distance intensity [SDI]), which we assign to stronger contributions from scalp hemodynamics to SDI and from cortical hemodynamics to DS?. Our findings suggest that scalp hemodynamic oscillations may be dominated by blood volume dynamics, whereas cortical hemodynamics may be dominated by blood flow velocity dynamics.     

Free fatty acids/triglycerides increase ocular and subcutaneous blood flow

Elevated plasma free fatty acids (FFA) induce skeletal muscle insulin resistance and impair endothelial function. The aim of this study was to characterize the acute hemodynamic effects of FFA in the eye and skin. A triglyceride (Intralipid 20%, 1.5 ml/min)/heparin (bolus: 200 IU; constant infusion rate: 0.2 IU. kg(-1). min(-1)) emulsion or placebo was administered to 10 healthy subjects. Measurements of pulsatile choroidal blood flow with laser interferometry, retinal blood flow with the blue field entoptic technique, peak systolic and end diastolic blood velocity (PSV, EDV) in the ophthalmic artery with Doppler sonography, and subcutaneous blood flow with laser Doppler flowmetry were performed during an euglycemic somatostatin-insulin clamp over 405 min. Plasma FFA/triglyceride elevation induced a rise in pulsatile choroidal blood flow by 25 +/- 3% (P < 0.001) and in retinal blood flow by 60 +/- 23% (P = 0.0125). PSV increased by 27 +/- 8% (P = 0.001), whereas EDV was not affected. Skin blood flow increased by 149 +/- 38% (P = 0.001). Mean blood pressure and pulse rate remained unchanged, whereas pulse pressure amplitude increased by 17 +/- 5% (P = 0.019). Infusion of heparin alone had no hemodynamic effect in the eye or skin. In conclusion, FFA/triglyceride elevation increases subcutaneous and ocular blood flow with a more pronounced effect in the retina than in the choroid, which may play a role for early changes of ocular perfusion in the insulin resistance syndrome.     

Differential effects of lower body negative pressure and upright tilt on splanchnic blood volume

Upright posture and lower body negative pressure (LBNP) both induce reductions in central blood volume. However, regional circulatory responses to postural changes and LBNP may differ. Therefore, we studied regional blood flow and blood volume changes in 10 healthy subjects undergoing graded lower-body negative pressure (-10 to -50 mmHg) and 8 subjects undergoing incremental head-up tilt (HUT; 20 degrees , 40 degrees , and 70 degrees ) on separate days. We continuously measured blood pressure (BP), heart rate, and regional blood volumes and blood flows in the thoracic, splanchnic, pelvic, and leg segments by impedance plethysmography and calculated regional arterial resistances. Neither LBNP nor HUT altered systolic BP, whereas pulse pressure decreased significantly. Blood flow decreased in all segments, whereas peripheral resistances uniformly and significantly increased with both HUT and LBNP. Thoracic volume decreased while pelvic and leg volumes increased with HUT and LBNP. However, splanchnic volume changes were directionally opposite with stepwise decreases in splanchnic volume with LBNP and stepwise increases in splanchnic volume during HUT. Splanchnic emptying in LBNP models regional vascular changes during hemorrhage. Splanchnic filling may limit the ability of the splanchnic bed to respond to thoracic hypovolemia during upright posture.     

Imaging photoplethysmography and videocapillaroscopy enable noninvasive study of zebrafish cardiovascular system functioning

We report on the noninvasive method for in vivo study of fish's cardiovascular system, that is, the heart and the structure of vessels that carry blood throughout the body. The proposed approach is based on combined photoplethysmographic and videocapillaroscopic microscopic imaging and enables noncontact two‐dimensional mapping of blood volume changes. We demonstrate that the obtained data allows precise measurements of heartbeat, blood flow velocity and other important parameters (see Videos S1 and S2 ). To validate the developed image processing technique, we have carried out multiple experiments on zebrafish—a well‐proven informative model organism widely used to understand cardiac development. The proposed approach may be effective for the study of cardiovascular system formation and functioning as well as the impact of various influencing factors on them.     

A deep‐learning‐based approach for noise reduction in high‐speed optical coherence Doppler tomography

Optical coherence Doppler tomography (ODT) increasingly attracts attention because of its unprecedented advantages with respect to high contrast, capillary‐level resolution and flow speed quantification. However, the trade‐off between the signal‐to‐noise ratio of ODT images and A‐scan sampling density significantly slows down the imaging speed, constraining its clinical applications. To accelerate ODT imaging, a deep‐learning‐based approach is proposed to suppress the overwhelming phase noise from low‐sampling density. To handle the issue of limited paired training datasets, a generative adversarial network is performed to implicitly learn the distribution underlying Doppler phase noise and to generate the synthetic data. Then a 3D based convolutional neural network is trained and applied for the image denoising. We demonstrate this approach outperforms traditional denoise methods in noise reduction and image details preservation, enabling high speed ODT imaging with low A‐scan sampling density.     

Mechanical aspects of heartbeat reversal in pupae of Manduca sexta

Pulsations of the dorsal vessel were investigated with new optocardiographic techniques based on the transmission and reflection of pulse-light through optic fibers. This noninvasive technique enabled simultaneous, in vivo multisensor recordings of the heartbeat without touching the pupal integument. There was a very regular heartbeat reversal with 3 distinctive phases: (a) a backward-oriented (retrograde) cardiac pulsation; (b) a forward-oriented (anterograde) pulsation with faster frequency; and (c) shorter or longer periods of temporary cardiac standstill that usually occurred after the termination of the anterograde phase. Occasionally, there were localized series of systolic cardiac contractions during the retrograde phase. Simultaneous recordings from the base and the tail of the abdomen revealed a reciprocal, "mirror image-like", quantitative relationship. The most intensive anterograde hemolymph flow occurred at the base while the most intensive retrograde flow occurred at the tail of the abdomen. The bi-directional switchovers of heartbeat (reversal) were occasionally associated with modifications during each of the unidirectional cardiac phases. Anterograde peristalsis showed a 2-fold higher frequency of pulsation in the thoracic aorta in comparison with the posterior parts of the heart. Thus, in addition to the "odd" peristaltic waves originating at the tail, there were intercallated "even" peristaltic waves originating in the middle of the abdomen. Both of them propagated hemolymph through the thoracic aorta into the head; the first waves took the hemolymph in from the distal end, while the second sucked it from the middle of the abdomen. The use of multiple optocardiographic sensors also enabled detection of cardiac pulsations on the opposite, ventral side of the body, within the ventral perineural sinus. The ventral side of the head showed only the presence of an anterograde pulse, whereas the ventral side of the tail exhibited a strong reciprocal retrograde phase and a very weak anterograde phase. These results explain why the existence of a periodic heartbeat reversal should be essential for circulatory functions at both extremities of the cylindrical insect body. In diapausing pupae, regular cycles of heartbeat reversal were substituted by prolonged periods of anterograde pulsation during the entire duration of bursts of CO2 release (average duration of the burst was 18-20 min, periodicity 5 to 18 h). The physiological nature of such feed-back correlation between heartbeat and metabolic CO2 production is not yet clear, because the anterograde heartbeat could be also induced by a number of nonspecific factors unrelated to CO2 (mechanical irritation, injury, injections, elevated temperature). During the postdiapause, developing pharate-adult stage, the correlation between CO2 and anterograde heartbeat completely disappeared. It has been concluded that regulation of insect heartbeat represents a highly coordinated, myogenic stereotype with inherent rhythmicity, which can be modified by a number of external and internal factors.     

Recognizing brain activities by functional near-infrared spectroscope signal analysis

Background

Noninvasive recording of movements caused by the heartbeat and the blood circulation is known as ballistocardiography. Several studies have shown the capability of a force plate to detect cardiac activity in the human body. The aim of this paper is to present a new method based on differential geometry of curves to handle multivariate time series obtained by ballistocardiographic force plate measurements.

Results

We show that the recoils of the body caused by cardiac motion and blood circulation provide a noninvasive method of displaying the motions of the heart muscle and the propagation of the pulse wave along the aorta and its branches. The results are compared with the data obtained invasively during a cardiac catheterization. We show that the described noninvasive method is able to determine the moment of a particular heart movement or the time when the pulse wave reaches certain morphological structure.

Conclusions

Monitoring of heart movements and pulse wave propagation may be used e.g. to estimate the aortic pulse wave velocity, which is widely accepted as an index of aortic stiffness with the application of predicting risk of heart disease in individuals. More extended analysis of the method is however needed to assess its possible clinical application.     

Brain‐derived neurotrophic factor genotype is associated with brain gray and white matter tissue volumes recovery in abstinent alcohol‐dependent individuals

Neuroimaging studies have linked the methionine (Met) allele of the brain‐derived neurotrophic factor (BDNF) gene to abnormal regional brain volumes in several psychiatric and neurodegenerative diseases. However, no neuroimaging studies assessed the effects of this allele on brain morphology in alcohol use disorders and its demonstrated change during abstinence from alcohol. Here we assessed the effects of the BDNF Val66Met (rs6265) polymorphism on regional brain tissue volumes and their recovery during short‐term abstinence in treatment‐seeking alcohol‐dependent individuals. 3D T1 weighted magnetic resonance images from 62 individuals were acquired at 1.5 T at one week of abstinence from alcohol; 41 of the participants were rescanned at 5 weeks of abstinence. The images were segmented into gray matter (GM), white matter (WM) and cerebrospinal fluid and parcellated into regional volumes. The BDNF genotype was determined from blood samples using the TaqMan technique. Alcohol‐dependent Val (Valine)/Met heterozygotes and Val homozygotes had similar regional brain volumes at either time point. However, Val homozygotes had significant GM volume increases, while Val/Met heterozygotes increased predominantly in WM volumes over the scan interval. Longitudinal increases in GM but not WM volumes were related to improvements in neurocognitive measures during abstinence. The findings suggest that functionally significant brain tissue volume recovery during abstinence from alcohol is influenced by BDNF genotype.     

Large‐scale ciliary reversal mediates capture of individual algal prey by Müller's larva

We documented capture of microalgal prey by several species of wild‐caught Müller's larvae of polyclad flatworms. To our knowledge, this is the first direct observation of feeding mechanism in this classical larval type. High‐speed video recordings showed that virtually all captures were mediated by large‐scale transient ciliary reversal over one or more portions of the main ciliary band corresponding to individual lobes or tentacles. Local ciliary beat reversals altered near‐field flow to suck parcels of food‐containing water mouthward. Many capture episodes entailed sufficient coordinated flow disruption that these compact‐bodied larvae tumbled dramatically. Similar behaviors were recorded in at least four distinct species, one of which corresponds to the ascidian‐eating polyclad Pseudoceros canadensis.