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Quantitative biomechanical characterization of pelvic supportive structures and functions in vivo is thought to provide insight into the pathophysiology of pelvic floor disorders including pelvic organ prolapse (POP). An innovative approach - vaginal tactile imaging - allows biomechanical mapping of the female pelvic floor to quantify tissue elasticity, pelvic support, and pelvic muscle functions. The objective of this study is to explore an extended set of 52 biomechanical parameters to characterize pelvic floor changes with age, parity, and subject weight for normal pelvic floor conditions. 42 subjects with normal pelvic conditions (no POP, no stress urinary incontinence) were included in the data analysis from an observational, case-controlled study. The Vaginal Tactile Imager (VTI) was used with an analytical software package to automatically calculate 52 biomechanical parameters for 8 VTI test procedures (probe insertion, elevation, rotation, Val-salva maneuver, voluntary muscle contractions in 2 planes, relaxation, and reflex contraction). The ranges, mean values, and standard deviations for all 52 VTI parameters were established. 12 VTI parameters were identified as statistically sen-sitive (p < 0.05; t-test) to the subject age; 9 parameters were identified as statistically sensitive (p < 0.05; t-test) to the subject parity; no sensitivity was found to subject weight. Among the 12 parameters sensitive to women’s age, 6 parameters show changes (decrease) in tissue elasticity and 6 parameters show weakness in pelvic muscle functions with age. Among the 9 parameters sensitive to parity, 5 parameters show changes (decrease) in tissue elasticity and 4 parameters show weakness in pelvic muscle functions after giving birth. The biomechanical mapping of the female pelvic floor with the VTI provides a unique set of parameters characterizing pelvic changes with age and parity. These objectively measurable biomechanical transformations of pelvic tissues, support structures, and functions may be used in future research and practical applications.

An amendment to this paper has been published and can be accessed via the original article.

The objective of this study was to evaluate if the Bone UltraSonic Scanner (BUSS) can detect osteoporosis in postmenopausal women. BUSS is an axial transmission multi-frequency ultrasonometer for acquisition of wave propagation profiles along the proximal anterior tibia. We derived 10 diagnostically significant BUSS parameters that were then compared with the DXA spine T-score, which was used in this study as the “gold standard” for the assessment of osteoporosis (T-score

We have previously hypothesized that the dissipation of mechanical energy of external impact is a fundamental function of skeletal muscle in addition to its primary function to convert chemical energy into mechanical energy. In this paper, a mathematical justification of this hypothesis is presented. First, a simple mechanical model, in which the muscle is considered as a simple Hookean spring, is considered. This analysis serves as an introduction to the consideration of a biomechanical model taking into account the molecular mechanism of muscle contraction, kinetics of myosin bridges, sarcomere dynamics, and tension of muscle fibers. It is shown that a muscle behaves like a nonlinear and adaptive spring tempering the force of impact and increasing the duration of the collision. The temporal profiles of muscle reaction to the impact as functions of the levels of muscle contraction, durations of the impact front, and the time constants of myosin bridges closing, are obtained. The absorption of mechanical shock energy is achieved due to the increased viscoelasticity of the contracting skeletal muscle. Controlling the contraction level allows for the optimization of the stiffness and viscosity of the muscle necessary for the protection of the joints and bones.

We develop a theory of the elasticity moduli and dissipative properties of a composite material: a phantom simulating muscle tissue anisotropy. The model used in the experiments was made of a waterlike polymer with embedded elastic filaments imitating muscle fiber. In contrast to the earlier developed phenomenological theory of the anisotropic properties of muscle tissue, here we obtain the relationship of the moduli with characteristic sizes and moduli making up the composite. We introduce the effective elasticity moduli and viscosity tensor components, which depend on stretching of the fibers. We measure the propagation velocity of shear waves and the shear viscosity of the model for regulated tension. Waves were excited by pulsed radiation pressure generated by modulated focused ultrasound. We show that with increased stretching of fibers imitating muscle contraction, an increase in both elasticity and viscosity takes place, and this effect depends on the wave propagation direction. The results of theoretical and experimental studies support our hypothesis on the protective function of stretched skeletal muscle, which protects bones and joints from trauma.

An acoustical handheld hydration monitor (HM) for assessing the water balance of the human body was developed. Dehydration is a critical public health problem. Many elderly over age of 65 are particularly vulnerable as are infants and young children. Given that dehydration is both preventable and reversible, the need for an easy-to-perform method for the detection of water imbalance is of the utmost clinical importance. The HM is based on an experimental fact that ultrasound velocity in muscle is a linear function of water content and can be referenced to the hydration status of the body. Studies on the validity of HM for the assessment of whole-body hydration status were conducted in the Appalachian State University, USA, on healthy young adults and on elderly subjects residing at an assisted living facility. The HM was able to track changes in total body water during periods of acute dehydration and rehydration in athletes and day-to-day and diurnal variability of hydration in elderly. Results of human studies indicate that HM has a potential to become an efficient tool for detecting abnormal changes in the body hydration status.

Atrial fibrillation is the most common cardiac arrhythmia. It is being effectively treated using the radiofrequency ablation (RFA) procedure, which destroys culprit tissue and creates scars that prevent the spread of abnormal electrical activity. Long-term success of RFA could be improved further if ablation lesions can be directly visualized during the surgery. We have shown that autofluorescence-based hyperspectral imaging (aHSI) can help to identify lesions based on spectral unmixing. We show that use of [Formula: see text]-means clustering, an unsupervised learning method, is capable of detecting RFA lesions without a priori knowledge of the lesions’ spectral characteristics. We also show that the number of spectral bands required for successful lesion identification can be significantly reduced, enabling the use of increased spectral bandwidth. Together, these findings can help with clinical implementation of a percutaneous aHSI catheter, since by reducing the number of spectral bands one can reduce hypercube acquisition and processing times, and by increasing the spectral width of individual bands one can collect more photons. The latter is of critical importance in low-light applications such as intracardiac aHSI. The ultimate goal of our studies is to help improve clinical outcomes for atrial fibrillation patients.

Rapidly improving protocols for the derivation of autologous cells from stem cell sources is a welcome development. However, there are many circumstances when off-the-shelf universally immunocompatible cells may be needed. Embryonic stem cells (ESCs) provide a unique opportunity to modify the original source of differentiated cells to minimize their rejection by nonautologous hosts.Immune rejection of nonautologous human embryonic stem cell (hESC) derivatives can be reduced by downregulating human leukocyte antigen (HLA) class I molecules, without affecting the ability of these cells to differentiate into specific lineages.Beta-2-microglobulin (B2M) expression was decreased by lentiviral transduction using human anti-HLA class I light-chain B2M short hairpin RNA. mRNA levels of B2M were decreased by 90% in a RUES2-modified hESC line, as determined by quantitative real time-polymerase chain reaction analysis. The transduced cells were selected under puromycin pressure and maintained in an undifferentiated state. The latter was confirmed by Oct4 and Nanog expression, and by the formation of characteristic round-shaped colonies. B2M downregulation led to diminished HLA-I expression on the cell surface, as determined by flow cytometry. When used as target cells in a mixed lymphocyte reaction assay, transduced hESCs and their differentiated derivatives did not stimulate allogeneic T-cell proliferation. Using a cardiac differentiation protocol, transduced hESCs formed a confluent layer of cardiac myocytes and maintained a low level of B2M expression. Transduced hESCs were also successfully differentiated into a hepatic lineage, validating their capacity to differentiate into multiple lineages.HLA-I depletion does not preclude hESC differentiation into cardiac or hepatic lineages. This methodology can be used to engineer tissue from nonautologous hESC sources with improved immunocompatibility.

Background To perform a colonoscopy, the endoscopist maneuvers the colonoscope through a series of loops by applying force to the insertion tube. Colonoscopy insertion techniques are operator dependent but have never been comprehensively quantified. Objective To determine whether the Colonoscopy Force Monitor (CFM), a device that continually measures force applied to the insertion tube, can identify different force application patterns among experienced endoscopists. Design Observational study of 6 experienced endoscopists performing routine diagnostic and therapeutic colonoscopy in 30 patients. Setting Outpatient ambulatory endoscopy center. Patients Adult male and female patients between 30 and 75 years of age undergoing routine colonoscopy. Interventions CFM monitoring of force applied to the colonoscope insertion tube during colonoscopy. Main Outcome Measurements Maximum and mean linear and torque force, time derivative of force, combined linear and torque vector force, and total manipulation time. Results The CFM demonstrates differences among endoscopists for maximum and average push/pull and mean torque forces, time derivatives of force, combined push/torque force vector, and total manipulation time. Endoscopists could be grouped by force application patterns. Limitations Only experienced endoscopists using conscious sedation in the patients were studied. Sample size was 30 patients. Conclusions This study demonstrates that CFM allows continuous force monitoring, characterization, and display of similarities and differences in endoscopic technique. CFM has the potential to facilitate training by enabling trainees to assess, compare, and quantify their techniques and progress.

BACKGROUND: Treatment of cardiac arrhythmias often involves ablating viable muscle tissue within or near islands of scarred myocardium. Yet, today there are limited means by which the boundaries of such scars can be visualized during surgery and distinguished from the sites of acute injury caused by radiofrequency (RF) ablation. OBJECTIVE: We sought to explore a hyperspectral imaging (HSI) methodology to delineate and distinguish scar tissue from tissue injury caused by RF ablation. METHODS: RF ablation of the ventricular surface of thoracotomized live rats was followed by a two-month animal recovery period. During a second surgery, new RF lesions were placed next to the scarred tissue from the previous ablation procedure. Myocardial infarction model was used as an alternative way to create scar tissue. RESULTS: Excitation-emission matrices acquired from the sites of RF lesions, scar region, and the surrounding unablated tissue revealed multiple spectral changes. These findings justified HSI of heart surface using illumination with 365nm ultraviolet light while acquiring spectral images from visible range. Autofluorescence-based HSI enabled to distinguish sites of RF lesions from scar or unablated myocardium in open chest rats. A pilot version of a percutaneous HSI catheter was used to demonstrate the feasibility of RF lesion visualization in atrial tissue of live pigs. CONCLUSIONS: Hyperspectral imaging based on changes in tissue autofluorescence is a highly effective tool for revealing, in vivo and with high spatial resolution, surface boundaries of a myocardial scar and discriminating it from areas of acute necrosis caused by RF ablation.

Ultrasound velocity in soft biological tissues is defined by the molecular composition of the tissues [1]. Water is the main molecular component of soft tissues and muscle water content is typically over 70%. Ultrasound velocity in muscle is a linear function of the water content [2]. Since the muscle is the main water reservoir of human body, measurements of ultrasound velocity in muscle may serve as a means for assessment of whole-body hydration status, which is an important physiological characteristic. A compact handheld device for measuring ultrasound velocity in muscle, named “hydration monitor” (HM), was developed by Artann Laboratories to assess the water balance in muscle. The objective of this study is to test whether the HM design provides the required accuracy of ultrasound velocity measurement to estimate muscle water content variations with sensitivity on the order of 1%.

The purpose of this study was to evaluate structural and optical properties of atrial tissue from common animal models and to compare it with human atria. We aimed to do this in a format that will be useful for development of better ablation tools and/or new means for visualizing atrial lesions. Human atrial tissue from clinically relevant age group was compared and contrasted with atrial tissue of large animal models commonly available for research purposes. These included pigs, sheep, dogs and cows. The presented data include area measurements of smooth atrial surface available for ablation and estimates of thickness of collagen and muscle for five different species. We also described methods to quantify presence of collagen and overall thickness of atrial wall. Provided information enables placement of atrial lesions to locations with clinically relevant atrial wall thickness and macroscopic structure ultimately helping investigators to develop better ablation and imaging tools. It also highlights the impact of collagen thickness on optical measurements and lesion visualization.

Mechanical imaging yields tissue elasticity map and provides quantitative characterization of a detected pathology. The changes in the surface stress patterns as a function of applied load provide information about the elastic composition and geometry of the underlying tissue structures. The objective of this study is the clinical evaluation of breast mechanical imager for breast lesion characterization and differentiation between benign and malignant lesions. The breast mechanical imager includes a probe with pressure sensor array, an electronic unit providing data acquisition from the pressure sensors and communication with a touch-screen laptop computer. We have developed an examination procedure and algorithms to provide assessment of breast lesion features such as hardness related parameters, mobility, and shape. A statistical Bayesian classifier was constructed to distinguish between benign and malignant lesions by utilizing all the listed features as the input. Clinical results for 179 cases, collected at four different clinical sites, have demonstrated that the breast mechanical imager provides a reliable image formation of breast tissue abnormalities and calculation of lesion features. Malignant breast lesions (histologically confirmed) demonstrated increased hardness and strain hardening as well as decreased mobility and longer boundary length in comparison with benign lesions. Statistical analysis of differentiation capability for 147 benign and 32 malignant lesions revealed an average sensitivity of 91.4% and specificity of 86.8% with a standard deviation of +/-6.1%. The area under the receiver operating characteristic curve characterizing benign and malignant lesion discrimination is 86.1% with the confidence interval ranging from 80.3 to 90.9%, with a significance level of P = 0.0001 (area = 50%). The multisite clinical study demonstrated the capability of mechanical imaging for characterization and differentiation of benign and malignant breast lesions. We hypothesize that the breast mechanical imager has the potential to be used as a cost effective device for cancer diagnostics that could reduce the benign biopsy rate, serve as an adjunct to mammography and to be utilized as a screening device for breast cancer detection.

The paper presents the theory of shear wave propagation in a “soft solid” material possessing anisotropy of elastic and dissipative properties. The theory is developed mainly for understanding the nature of the low-frequency acoustic characteristics of skeletal muscles, which carry important diagnostic information on the functional state of muscles and their pathologies. It is shown that the shear elasticity of muscles is determined by two independent moduli. The dissipative properties are determined by the fourth-rank viscosity tensor, which also has two independent components. The propagation velocity and attenuation of shear waves in muscle depend on the relative orientation of three vectors: the wave vector, the polarization vector, and the direction of muscle fiber. For one of the many experiments where attention was distinctly focused on the vector character of the wave process, it was possible to make a comparison with the theory, estimate the elasticity moduli, and obtain agreement with the angular dependence of the wave propagation velocity predicted by the theory.

We hypothesize that dissipation of mechanical energy of external impact to absorb mechanical shock is a fundamental function of skeletal muscle in addition to its primary function to convert chemical energy into mechanical energy. In physical systems, the common mechanism for absorbing mechanical shock is achieved with the use of both elastic and viscous elements and we hypothesize that the viscosity of the skeletal muscle is a variable parameter which can be voluntarily controlled by changing the tension of the contracting muscle. We further hypothesize that an ability of muscle to absorb shock has been an important factor in biological evolution, allowing the life to move from the ocean to land, from hydrodynamic to aerodynamic environment with dramatically different loading conditions for musculoskeletal system. The ability of muscle to redistribute the energy of mechanical shock in time and space and unload skeletal joints is of key importance in physical activities. We developed a mathematical model explaining the absorption of mechanical shock energy due to the increased viscosity of contracting skeletal muscles. The developed model, based on the classical theory of sliding filaments, demonstrates that the increased muscle viscosity is a result of the time delay (or phase shift) between the mechanical impact and the attachment/detachment of myosin heads to binding sites on the actin filaments. The increase in the contracted muscle’s viscosity is time dependent. Since the forward and backward rate constants for binding the myosin heads to the actin filaments are on the order of 100 s 1 , the viscosity of the contracted muscle starts to significantly increase with an impact time greater than 0.01 s. The impact time is one of the key parameters in generating destructive stress in the colliding objects. In order to successfully dampen a short high power impact, muscles must first slow it down to engage the molecular mechanism of muscle viscosity. Muscle carries out two functions, acting first as a nonlinear spring to slow down impact and second as a viscous damper to absorb the impact. Exploring the ability of muscle to absorb mechanical shock may shed light to many problems of medical biomechanics and sports medicine. Currently there are no clinical devices for real-time quantitative assessment of viscoelastic properties of contracting muscles in vivo. Such assessment may be important for diagnosis and monitoring of treatment of various muscle disorders such as muscle dystrophy, motor neuron diseases, inflammatory and metabolic myopathies and many more.

Background: Bisphenol A (BPA) is used to produce polycarbonate plastics and epoxy resins that are widely used in everyday products, such as food and beverage containers, toys, and medical devices. Human biomonitoring studies have suggested that a large proportion of the population may be exposed to BPA. Recent epidemiological studies have reported correlations between increased urinary BPA concentrations and cardiovascular disease, yet the direct effects of BPA on the heart are unknown. Objectives: The goal of our study was to measure the effect of BPA (0.1–100 μM) on cardiac impulse propagation ex vivo using excised whole hearts from adult female rats. Methods: We measured atrial and ventricular activation times during sinus and paced rhythms using epicardial electrodes and optical mapping of transmembrane potential in excised rat hearts exposed to BPA via perfusate media. Atrioventricular activation intervals and epicardial conduction velocities were computed using recorded activation times. Results: Cardiac BPA exposure resulted in prolonged PR segment and decreased epicardial conduction velocity (0.1–100 μM BPA), prolonged action potential duration (1–100 μM BPA), and delayed atrioventricular conduction (10–100 μM BPA). These effects were observed after acute exposure (≤ 15 min), underscoring the potential detrimental effects of continuous BPA exposure. The highest BPA concentration used (100 μM) resulted in prolonged QRS intervals and dropped ventricular beats, and eventually resulted in complete heart block. Conclusions: Our results show that acute BPA exposure slowed electrical conduction in excised hearts from female rats. These findings emphasize the importance of examining BPA’s effect on heart electrophysiology and determining whether chronic in vivo exposure can cause or exacerbate conduction abnormalities in patients with preexisting heart conditions and in other high-risk populations. Citation: Posnack NG, Jaimes R III, Asfour H, Swift LM, Wengrowski AM, Sarvazyan N, Kay MW. 2014. Bisphenol A exposure and cardiac electrical conduction in excised rat hearts. Environ Health Perspect 122:384–390; http://dx.doi.org/10.1289/ehp.1206157

This article considers the use of autologous stem cell-derived cardiomyocytes as a novel means to aid venous return. The approach consists of creating external cuffs of engineered heart tissue around vein segments with incompetent or poorly competent valves. The engineered heart tissue cuff prevents distention of the impaired vein segments and aids unidirectional flow by its rhythmic contractions. There appear to be no fundamental limitations to this approach as feasibility of all of the individual components has already been shown. Here, we underline the clinical need for novel ways to treat chronic deep venous insufficiency, review previous research that enabled this approach, consider potential designs of engineered heart tissue cuffs, and outline its advantages and future challenges.

The present study examines the association of the changes in ultrasound velocity measured at 1 MHz using 1.5 micros duration tone burst in the human soleus muscle in vivo with several pathologies including patients with chronic renal failure (CRF) and disorders of the cardiovascular system. Total 127 subjects were investigated, with approximately equal number of male and female subjects uniformly distributed by age, from 15 to 70 years old. Since molecular composition of the tissue is thought to have greater effect on the bulk ultrasound velocity, potential contribution of both water and fat, two main variable components of a muscle, were taken into account. Observed negative correlation of ultrasound velocity with the body mass index was considered a result of an elevated fat content. Based on the obtained data, presence of leg edemas results in a measurably lower ultrasound velocity in the soleus muscle. Unless patients had visibly detected leg edema, no difference between healthy individuals, patients with chronic heart failure, or CRF was found. Despite relatively high individual variations in velocity, ranging from 1530 to 1615 m/s, a statistically significant gender correlated difference between average values of the velocity was observed. No dependence of velocity on subject age was detected. An indirect confirmation of the muscle fluid homeostasis was revealed in patients with CRF undergoing hemodialysis procedure. After hemodialysis, a significantly smaller increase (0.3% in average) of ultrasound velocity in the soleus muscle was observed than otherwise could be expected if a uniform relative loss of total body fluids was assumed (1-1.3%). In general, the study findings set a premise for using ultrasound velocity as a potential quantitative parameter for edema assessment.

Tissue water content is an important diagnostic parameter that can be used for estimation of water loss in muscles such as common dehydration during high endurance exercises. It could be also applied for evaluation of the increased fluids content in the tissue caused by the variety of pathological conditions or edemas. Ultrasonic method for tissue water content monitoring is based on the premise that the speed of a bulk or compression sound wave is determined mainly by the molecular content of the tissue. Most soft tissues, including muscles that consist of about 70-80% water, exhibit shift of the ultrasound velocity associated with the change in their water content. In the present paper, we tested the feasibility of assessing changes in tissue water content by measurements of ultrasound velocity in ex vivo animal muscle tissues. An increase in the ultrasound velocity correlated with the volumetric water loss in the tissue was observed when other tissue components (proteins, fat) remained constant. Possibility to assess muscle dehydration with 1% accuracy was confirmed in model dehydration experiments, where ultrasound velocity slope of about 3 m/s per 1% of water loss was revealed at measurement error less than 2 m/s. Hence, the ultrasonic approach can provide basis for a convenient, lightweight system in sports medicine for monitoring total body hydration during long-term endurance exercise in hot conditions, as well as for edemas monitoring and other medical applications.

Time-reversal acoustics is an effective way of focusing ultrasound deep inside heterogeneous media such as biological tissues. Convection-enhanced delivery is a method of delivering drugs into the brain by infusing them directly into the brain interstitium. These two technologies are combined in a focusing system that uses a “smart needle” to simultaneously infuse fluid into the brain and provide the necessary feedback for focusing ultrasound using time-reversal acoustics. The effects of time-reversal acoustics-focused ultrasound on the spatial distribution of infused low- and high-molecular weight tracer molecules are examined in live, anesthetized rats. Results show that exposing the rat brain to focused ultrasound significantly increases the penetration of infused compounds into the brain. The addition of stabilized microbubbles enhances the effect of ultrasound exposure.

Up until about two decades ago acoustic imaging and ultrasound imaging were synonymous. The term “ultrasonography,” or its abbreviated version “sonography” meant an imaging modality based on the use of ultrasonic compressional bulk waves. Since the 1990s numerous acoustic imaging modalities started to emerge based on the use of a different mode of acoustic wave: shear waves. It was demonstrated that imaging with these waves can provide very useful and very different information about the biological tissue being examined. We will discuss physical basis for the differences between these two basic modes of acoustic waves used in medical imaging and analyze the advantages associated with shear acoustic imaging. A comprehensive analysis of the range of acoustic wavelengths, velocities, and frequencies that have been used in different imaging applications will be presented. We will discuss the potential for future shear wave imaging applications.

Radiofrequency ablation (RFA) is a widely used treatment for atrial fibrillation, the most common cardiac arrhythmia. Here, we explore autofluorescence hyperspectral imaging (aHSI) as a method to visualize RFA lesions and interlesional gaps in the highly collagenous left atrium. RFA lesions made on the endocardial surface of freshly excised porcine left atrial tissue were illuminated by UV light (365 nm), and hyperspectral datacubes were acquired over the visible range (420–720 nm). Linear unmixing was used to delineate RFA lesions from surrounding tissue, and lesion diameters derived from unmixed component images were quantitatively compared to gross pathology. RFA caused two consistent changes in the autofluorescence emission profile: a decrease at wavelengths below 490 nm (ascribed to a loss of endogenous NADH) and an increase at wavelengths above 490 nm (ascribed to increased scattering). These spectral changes enabled high resolution, in situ delineation of RFA lesion boundaries without the need for additional staining or exogenous markers. Our results confirm the feasibility of using aHSI to visualize RFA lesions at clinically relevant locations. If integrated into a percutaneous visualization catheter, aHSI would enable widefield optical surgical guidance during RFA procedures and could improve patient outcome by reducing atrial fibrillation recurrence.

Persistent atrial fibrillation is commonly treated using an endoscopic catheter to eliminate anomalous sources of cardiac activity via thermal ablation. However, the procedure lacks real-time feedback. Newly developed radiofrequency ablation (RFA) catheters include a fiberoptic bundle through which visual information of tissue conditions may be collected offering an opportunity to reveal subtle differences in tissue physiology. Currently little is known about the spectral changes caused by RFA. We hypothesized that by comparing spectral changes in various areas in the heart before and after RFA, optical signatures can be used to distinguish healthy cardiac tissue from thermally ablated tissue. Excitation emission matrices (EEM) were acquired from excised porcine hearts (300-600nm). Distinct EEMs were collected from the endocardium of the left atria, ventricle, and aorta. Additionally, the fluorescence and reflectance profiles of each tissue were altered by thermal ablation. In the ventricular muscle, a reduction in the NADH fluorescence peak (360/460nm excitation/emission maxima) was most prominent. While in the aorta, collagen and elastin fluorescence peaks fused and broadened upon ablation. Changes in atrial tissue included a drop in NADH fluorescence and an overall increase in reflectance. The latter is likely caused by thermal coagulation of heme-containing proteins such as myoglobin and a weaker absorption within the Soret band. We concluded that optical signals revealed by EEMs offer quantitative information that can be used to develop diagnostic catheters, including hyperspectral imaging protocols to discern spectral changes elicited by RFA treatment.

New diagnostic catheters can be developed by delivering and acquiring light through a small fiberoptic bundle. This can provide useful real time feedback guidance to observe tissue damage caused by thermal injury used to treat cardiac fibrillation. Yet, little is known about the exact spectral changes caused by radiofrequency ablation (RFA) in different types of cardiac tissue. We hypothesized that the most sensitive optical ranges for characterizing thermal injury can be revealed by comparing spectral information from different areas of the heart before and after RF ablation. Freshly excised porcine hearts were used to acquire and analyze excitation emission matrices (EEMs, 300-600nm) from ventricular muscle, endocardium of the left atria, and aorta. Each type of tissue exhibited distinct EEMs that underwent highly reproducible changes in fluorescence absorption and reflectance upon RF ablation. Specifically, RFA resulted in a reduction of the NADH fluorescence peak in ventricular muscle EEMs (360/460nm excitation/emission maxima). It also led to a broadening and fusing of collagen and elastin fluorescence peaks in the aorta and it had a combined effect on left atrial tissue. RFA led to an increase in specular and diffuse reflectance (seen as higher amplitude and width of the EEM diagonal line) in all three tissue types. Thermal coagulation of heme-containing proteins, including different forms of myoglobin, led to a weaker absorption in the Soret band range (410-430nm). The latter was particularly noticeable in ventricular tissue but was also significant in the left atrial tissue. EEMs can provide a wealth of quantitative information, including types of optical measurements and specific wavelength ranges that can help to develop new diagnostic tools, including hyperspectral imaging techniques, to probe for spectral changes during RF ablation of different parts of the heart.

Toward the end of the 20th century, a new technology for measuring and visualizing soft tissue viscoelastic characteristics evolved. This modality is called elastography, or elasticity imaging. Tactile imaging, a form of elastography, translates the sense of touch into a digital image and allows assessment of tissue elasticity and muscle strength. The female pelvic organs likely have varying viscoelastic characteristics across physiological states, healthy and diseased conditions. This makes the pelvic floor tissues amenable to evaluation by tactile imaging techniques, opening up new possibilities for the quantitative biomechanical assessment and monitoring of pelvic floor conditions. The vaginal tactile imager offers an opportunity to assess the vaginal support along the entire length of the anterior, posterior, and lateral vaginal walls at rest, with manually applied deflection pressures and with voluntary, involuntary muscle contraction, involuntary relaxation, and Valsalva maneuver. This allows a large body of measurements to evaluate individual variations in support defects as well as identify specific potential markers to measure tissue properties and muscle function in patients with pelvic organ prolapse and stress urinary incontinence. The proposed approach may help further differentiate the types of pelvic floor conditions, their underlying severity, and understand how to tailor treatments for the individual patient by the most effective manner.

Blebbistatin is a recently discovered myosin II inhibitor. It is rapidly becoming a compound of choice to reduce motion artifacts during cardiac optical mapping, as well as to study cell motility and cell invasion. Although blebbistatin has a number of advantages over other electromechanical uncouplers, many of its properties have yet to be addressed. Here we describe several methodological issues associated with the use of blebbistatin, including its spectral properties, reversibility, and its effect on tissue metabolic state. We show that if precautions are not taken, perfusion with blebbistatin may result in blebbistatin precipitate that accumulates in the vasculature. Although such precipitate is fluorescent, it is not detectable within wavelength bands that are typically used for transmembrane voltage fluorescence imaging (i.e., emission wavelengths >600 nm). Therefore, blockage of the microcirculation by blebbistatin may cause data misinterpretation in studies that use voltage-sensitive dyes. Blebbistatin may also impact imaging of green fluorophores due to the spectral shift it causes in endogenous tissue fluorescence. 3D excitation–emission matrices of blebbistatin in precipitate form and in various solutions (DMSO, water, and 1 % aqueous albumin) revealed significant changes in the fluorescence of this molecule in different environments. Finally, we examined the reversibility of blebbistatin’s uncoupling effect on cardiac contraction. Our findings provide important new information about the properties of this myosin II inhibitor, which will aid in the proper design and interpretation of studies that use this compound.

Transplantation studies in mice and rats have shown that human embryonic-stem-cell-derived cardiomyocytes (hESC-CMs) can improve the function of infarcted hearts, but two critical issues related to their electrophysiological behaviour in vivo remain unresolved. First, the risk of arrhythmias following hESC-CM transplantation in injured hearts has not been determined. Second, the electromechanical integration of hESC-CMs in injured hearts has not been demonstrated, so it is unclear whether these cells improve contractile function directly through addition of new force-generating units. Here we use a guinea-pig model to show that hESC-CM grafts in injured hearts protect against arrhythmias and can contract synchronously with host muscle. Injured hearts with hESC-CM grafts show improved mechanical function and a significantly reduced incidence of both spontaneous and induced ventricular tachycardia. To assess the activity of hESC-CM grafts in vivo, we transplanted hESC-CMs expressing the genetically encoded calcium sensor, GCaMP3 (refs 4, 5). By correlating the GCaMP3 fluorescent signal with the host ECG, we found that grafts in uninjured hearts have consistent 1:1 host–graft coupling. Grafts in injured hearts are more heterogeneous and typically include both coupled and uncoupled regions. Thus, human myocardial grafts meet physiological criteria for true heart regeneration, providing support for the continued development of hESC-based cardiac therapies for both mechanical and electrical repair.