Distal planar rotary scanner for endoscopic optical coherence tomography.

Optical coherence tomography (OCT) is becoming a more common endoscopic imaging modality for detecting and treating disease given its high resolution and image quality. To use OCT for 3-dimensional imaging of small lumen, embedding an optical scanner at the distal end of an endoscopic probe for circumferential scanning the probing light is a promising way to implement high-quality imaging unachievable with the conventional method of revolving an entire probe. To this end, the present work proposes a hollow and planar micro rotary actuator for its use as an endoscopic distal scanner. A miniaturized design of this ferrofluid-assisted electromagnetic actuator is prototyped to act as a full 360° optical scanner, which is integrated at the tip of a fiber-optic probe together with a gradient-index lens for use with OCT. The scanner is revealed to achieve a notably improved dynamic performance that shows a maximum speed of 6500 rpm, representing 325% of the same reported with the preceding design, while staying below the thermal limit for safe in-vivo use. The scanner is demonstrated to perform real-time OCT using human fingers as live tissue samples for the imaging tests. The acquired images display no shadows from the electrical wires to the scanner, given its hollow architecture that allows the probing light to pass through the actuator body, as well as the quality high enough to differentiate the dermis from the epidermis while resolving individual sweat glands, proving the effectiveness of the prototyped scanner design for endoscopic OCT application.

Intrarenal Pressure Measured Using a Novel Flexible Ureteroscope with Pressure Sensing Capabilities: A Study of the Effects of Ureteral Access Sheath, Irrigation, and Working Channel Accessories.

Introduction: Intrarenal pressure is emerging as an important monitoring parameter during flexible ureteroscopy (fURS). We measure how intraoperative conditions affect intrarenal pressure using a novel pressure sensing ureteroscope. Methods: The LithoVue Elite (LVE) single-use digital flexible ureteroscope (Boston Scientific) is the first commercial ureteroscope that senses pressure at its tip. LVE was used in a porcine model to measure intrarenal pressure with and without a ureteral access sheath (UAS) with various sizes and placement locations, irrigation methods, and working channel accessories. LVE pressure accuracy was measured in a bench model. This abstract shows the least-square means from multiway analysis of variances used for analysis. Results: Intrarenal pressures were the highest without a UAS (64 mm Hg), followed by the 11/13 UAS (51 mm Hg) and the 12/14 and 13/15, which were not statistically different (39-40 mm Hg). The pressures were highest with UASs placed at the ureteropelvic junction (61 mm Hg), and lowest if placed in the renal pelvis (24 mm Hg). Irrigation methods showed the highest pressures with syringe (57 mm Hg), while irrigation bags (pressurized at 150-300 mm Hg) produced 43 to 46 mm Hg and 25 mm Hg when applied with 80 cm of gravity. Placing a 200 µm laser fiber reduced pressures from 44 to 41 mm Hg. Finally, the bench model showed that LVE was 96% accurate up to 300 mm Hg. Conclusion: Intrarenal pressure significantly varied based on UAS sizes, placement, and irrigation methods. Accordingly, fURS with LVE is poised to be an invaluable tool for clinical decision-making and future studies of intrarenal pressure.

Switch mode capacitive pressure sensors.

Switch mode capacitive pressure sensors are proposed as a new class of microfabricated devices that transform pressure into a mechanically switching capacitance to form an analog-to-digital signal with zero power, high sensitivity, and a high signal-to-noise ratio. A pressure-sensitive gold membrane suspended over a capacitive cavity makes ohmic contact with patterned gold leads on the substrate, closing circuits to fixed on-chip capacitors outside the cavity and leading to significant step responses. This function is achieved by allocating the switch leads on the part of the counter electrode area, while the remaining area is used for touch mode analog capacitive sensing. The sensor microchip is prototyped through a novel design approach to surface micromachining that integrates micro-Tesla valves for vacuum sealing the sensor cavity, showing an unprecedented response to applied pressure. For a gauge pressure range of 0-120 mmHg, the sensor exhibits an increase of 13.21 pF with resultant switch events, each of which ranges from 2.53-3.96 pF every 12-38 mmHg, in addition to the touch mode linear capacitive increase between switches. The equivalent sensitivity is 80-240 fF/mmHg, which is 11-600× more than commercial and reported touch mode sensors operating in similar pressure ranges. The sensor is further demonstrated for wireless pressure tracking by creating a resonant tank with the sensor, showing a 32.5-101.6 kHz/mmHg sensitivity with frequency jumps led by the switch events. The developed sensor, with its promising performance, offers new application opportunities in a variety of device areas, including health care, robotics, industrial control, and environmental monitoring.

Nonlinear Actomyosin Elasticity in Muscle?

Cyclic interactions between myosin II motor domains and actin filaments that are powered by turnover of ATP underlie muscle contraction and have key roles in motility of nonmuscle cells. The elastic characteristics of actin-myosin cross-bridges are central in the force-generating process, and disturbances in these properties may lead to disease. Although the prevailing paradigm is that the cross-bridge elasticity is linear (Hookean), recent single-molecule studies suggest otherwise. Despite convincing evidence for substantial nonlinearity of the cross-bridge elasticity in the single-molecule work, this finding has had limited influence on muscle physiology and physiology of other ordered cellular actin-myosin ensembles. Here, we use a biophysical modeling approach to close the gap between single molecules and physiology. The model is used for analysis of available experimental results in the light of possible nonlinearity of the cross-bridge elasticity. We consider results obtained both under rigor conditions (in the absence of ATP) and during active muscle contraction. Our results suggest that a wide range of experimental findings from mechanical experiments on muscle cells are consistent with nonlinear actin-myosin elasticity similar to that previously found in single molecules. Indeed, the introduction of nonlinear cross-bridge elasticity into the model improves the reproduction of key experimental results and eliminates the need for force dependence of the ATP-induced detachment rate, consistent with observations in other single-molecule studies. The findings have significant implications for the understanding of key features of actin-myosin-based production of force and motion in living cells, particularly in muscle, and for the interpretation of experimental results that rely on stiffness measurements on cells or myofibrils.

Sarcomere Stiffness during Stretching and Shortening of Rigor Skeletal Myofibrils.

In this study, we measured the stiffness of skeletal muscle myofibrils in rigor. Using a custom-built atomic force microscope, myofibrils were first placed in a rigor state then stretched and shortened at different displacements (0.1-0.3 µm per sarcomere) and nominal speeds (0.4 and 0.8 µm/s). During stretching, the myofibril stiffness was independent of both displacement and speed (average of 987 nN/µm). During shortening, the myofibril stiffness was independent of displacement, but dependent on speed (1234 nN/µm at 0.4 µm/s; 1106 nN/µm at 0.8 µm/s). Furthermore, the myofibril stiffness during shortening was greater than that during stretching and the difference depended on speed (31% at 0.4 µm/s; 8% at 0.8 µm/s). The results suggest that the myofibrils exhibit nonlinear viscoelastic properties that may be derived from myofibril filaments, similar to what has been observed in muscle fibers.

Residual force enhancement is regulated by titin in skeletal and cardiac myofibrils.

KEY POINTS: When a skeletal muscle is stretched while it contracts, the muscle produces a relatively higher force than the force from an isometric contraction at the same length: a phenomenon referred to as residual force enhancement. Residual force enhancement is puzzling because it cannot be directly explained by the classical force-length relationship and the sliding filament theory of contraction, the main paradigms in the muscle field. We used custom-built instruments to measure residual force enhancement in skeletal myofibrils, and, for the first time, in cardiac myofibrils. Our data report that residual force enhancement is present in skeletal muscles, but not cardiac muscles, and is regulated by the different isoforms of the titin protein filaments. ABSTRACT: When a skeletal muscle contracts isometrically, the muscle produces a force that is relative to the final isometric sarcomere length (SL). However, when the same final SL is reached by stretching the muscle while it contracts, the muscle produces a relatively higher force: a phenomenon commonly referred to as residual force enhancement. In this study, we investigated residual force enhancement in rabbit skeletal psoas myofibrils and, for the first time, cardiac papillary myofibrils. A custom-built atomic force microscope was used in experiments that stretched myofibrils before and after inhibiting myosin and actin interactions to determine whether the different cardiac and skeletal titin isoforms regulate residual force enhancement. At SLs ranging from 2.24 to 3.13 µm, the skeletal myofibrils enhanced the force by an average of 9.0%, and by 29.5% after hindering myosin and actin interactions. At SLs ranging from 1.80 to 2.29 µm, the cardiac myofibrils did not enhance the force before or after hindering myosin and actin interactions. We conclude that residual force enhancement is present only in skeletal muscles and is dependent on the titin isoforms.

Ingestible gastrointestinal sampling devices: state-of-the-art and future directions.

Despite the significant contribution of gastrointestinal diseases to the global disease burden and the increasing recognition of the role played by the intestinal microbiota in human health and disease states, conventional methods of exploring and collecting samples from the gastrointestinal tract remain invasive, resource intensive, and often unable to capture all the information contained in these heterogeneous samples. A new class of gastrointestinal sampling capsules is emerging in the literature, which contains the components required for an autonomous intra-luminal device and preserves the spatial and temporal information of the gastrointestinal samples. In this paper, we identify the primary design requirements for gastrointestinal sampling capsules, and we review the state-of-the-art for different components and functionalities. We also suggest two design concepts, and we highlight future directions for this class of biomedical devices.

Forces measured with micro-fabricated cantilevers during actomyosin interactions produced by filaments containing different myosin isoforms and loop 1 structures.

BACKGROUND: There is evidence that the actin-activated ATP kinetics and the mechanical work produced by muscle myosin molecules are regulated by two surface loops, located near the ATP binding pocket (loop 1), and in a region that interfaces with actin (loop 2). These loops regulate force and velocity of contraction, and have been investigated mostly in single molecules. There is a lack of information of the work produced by myosin molecules ordered in filaments and working cooperatively, which is the actual muscle environment. METHODS: We use micro-fabricated cantilevers to measure forces produced by myosin filaments isolated from mollusk muscles, skeletal muscles, and smooth muscles containing variations in the structure of loop 1 (tonic and phasic myosins). We complemented the experiments with in-vitro assays to measure the velocity of actin motility. RESULTS: Smooth muscle myosin filaments produced more force than skeletal and mollusk myosin filaments when normalized per filament overlap. Skeletal muscle myosin propelled actin filaments in a higher sliding velocity than smooth muscle myosin. The values for force and velocity were consistent with previous studies using myosin molecules, and suggest a close correlation with the myosin isoform and structure of surface loop 1. GENERAL SIGNIFICANCE: The technique using micro-fabricated cantilevers to measure force of filaments allows for the investigation of the relation between myosin structure and contractility, allowing experiments to be conducted with an array of different myosin isoforms. Using the technique we observed that the work produced by myosin molecules is regulated by amino-acid sequences aligned in specific loops.

Forces measured with micro-fabricated cantilevers during actomyosin interactions produced by filaments containing different myosin isoforms and loop 1 structures.

BACKGROUND: There is evidence that the actin-activated ATP kinetics and the mechanical work produced by muscle myosin molecules are regulated by two surface loops, located near the ATP binding pocket (loop 1), and in a region that interfaces with actin (loop 2). These loops regulate force and velocity of contraction, and have been investigated mostly in single molecules. There is a lack of information of the work produced by myosin molecules ordered in filaments and working cooperatively, which is the actual muscle environment. METHODS: We use micro-fabricated cantilevers to measure forces produced by myosin filaments isolated from mollusk muscles, skeletal muscles, and smooth muscles containing variations in the structure of loop 1 (tonic and phasic myosins). We complemented the experiments with in-vitro assays to measure the velocity of actin motility. RESULTS: Smooth muscle myosin filaments produced more force than skeletal and mollusk myosin filaments when normalized per filament overlap. Skeletal muscle myosin propelled actin filaments in a higher sliding velocity than smooth muscle myosin. The values for force and velocity were consistent with previous studies using myosin molecules, and suggest a close correlation with the myosin isoform and structure of surface loop 1. GENERAL SIGNIFICANCE: The technique using micro-fabricated cantilevers to measure force of filaments allows for the investigation of the relation between myosin structure and contractility, allowing experiments to be conducted with an array of different myosin isoforms. Using the technique we observed that the work produced by myosin molecules is regulated by amino-acid sequences aligned in specific loops.