Modeling thick filament activation suggests a molecular basis for force depression.

Multiscale models aiming to connect muscle's molecular and cellular function have been difficult to develop, in part due to a lack of self-consistent multiscale data. To address this gap, we measured the force response from single, skinned rabbit psoas muscle fibers to ramp shortenings and step stretches performed on the plateau region of the force-length relationship. We isolated myosin from the same muscles and, under similar conditions, performed single-molecule and ensemble measurements of myosin's ATP-dependent interaction with actin using laser trapping and in vitro motility assays. We fit the fiber data by developing a partial differential equation model that includes thick filament activation, whereby an increase in force on the thick filament pulls myosin out of an inhibited state. The model also includes a series elastic element and a parallel elastic element. This parallel elastic element models a titin-actin interaction proposed to account for the increase in isometric force after stretch (residual force enhancement). By optimizing the model fit to a subset of our fiber measurements, we specified seven unknown parameters. The model then successfully predicted the remainder of our fiber measurements and also our molecular measurements from the laser trap and in vitro motility. The success of the model suggests that our multiscale data are self-consistent and can serve as a testbed for other multiscale models. Moreover, the model captures the decrease in isometric force observed in our muscle fibers after active shortening (force depression), suggesting a molecular mechanism for force depression, whereby a parallel elastic element combines with thick filament activation to decrease the number of cycling cross-bridges.

Modeling Thick Filament Activation Suggests a Molecular Basis for Force Depression.

Multiscale models aiming to connect muscle's molecular and cellular function have been difficult to develop, in part, due to a lack of self-consistent multiscale data. To address this gap, we measured the force response from single skinned rabbit psoas muscle fibers to ramp shortenings and step stretches performed on the plateau region of the force-length relationship. We isolated myosin from the same muscles and, under similar conditions, performed single molecule and ensemble measurements of myosin's ATP-dependent interaction with actin using laser trapping and in vitro motility assays. We fit the fiber data by developing a partial differential equation model that includes thick filament activation, whereby an increase in force on the thick filament pulls myosin out of an inhibited state. The model also includes a series elastic element and a parallel elastic element. This parallel elastic element models a titin-actin interaction proposed to account for the increase in isometric force following stretch (residual force enhancement). By optimizing the model fit to a subset of our fiber measurements, we specified seven unknown parameters. The model then successfully predicted the remainder of our fiber measurements and also our molecular measurements from the laser trap and in vitro motility. The success of the model suggests that our multiscale data are self-consistent and can serve as a testbed for other multiscale models. Moreover, the model captures the decrease in isometric force observed in our muscle fibers after active shortening (force depression), suggesting a molecular mechanism for force depression, whereby a parallel elastic element combines with thick filament activation to decrease the number of cycling cross-bridges.

Tibiofemoral loss of contact area but no changes in peak pressures after meniscectomy in a Lapine in vivo quadriceps force transfer model.

PURPOSE: The menisci are thought to modulate load transfer and to absorb shocks in the knee joint. No study has experimentally measured the meniscal functions in the intact, in vivo joint loaded by physiologically relevant muscular contractions. METHODS: Right knee joints of seven New Zealand white rabbits were loaded using isometric contractions of the quadriceps femoris muscles controlled by femoral nerve stimulation. Isometric knee extensor torques at the maximal and two submaximal force levels were performed at knee angles of 70°, 90°, 110°, and 130°. Patellofemoral and tibiofemoral contact areas and pressure distributions were measured using Fuji Presensor film inserted above and below the menisci and also with the menisci removed. RESULTS: Meniscectomy was associated with a decrease in tibiofemoral contact area ranging from 30 to 70% and a corresponding increase in average contact pressures. Contact areas measured below the menisci were consistently larger than those measured on top of the menisci. Contact areas in the patellofemoral joint (PFJ), and peak pressures in tibiofemoral and PFJs, were not affected by meniscectomy. Contact areas and peak pressures in all joints depended crucially on knee joint angle and quadriceps force: The more flexed the knee joint was, the larger were the contact areas and the higher were the peak pressures. CONCLUSIONS: In agreement with the literature, removal of the menisci was associated with significant decreases in tibiofemoral contact area and corresponding increases in average contact pressures, but surprisingly, peak pressures remained unaffected, indicating that the function of the menisci is to distribute loads across a greater contact area.

Internal Carotid Artery Strains During High-Speed, Low-Amplitude Spinal Manipulations of the Neck.

OBJECTIVE: The primary objective of this study was to quantify the strains applied to the internal carotid artery (ICA) during neck spinal manipulative treatments and range of motion (ROM)/diagnostic testing of the head and neck. METHODS: Strains of the ICA (n = 12) were measured in 6 fresh, unembalmed cadaveric specimens using sonomicrometry. Peak and average strains of the ICA obtained during cervical spinal manipulations given by experienced doctors of chiropractic were compared with the corresponding strains obtained during ROM and diagnostic testing of the head and neck. RESULTS: Peak and average strains of the ICA for cervical spinal manipulative treatments were significantly smaller (P < .001) than the corresponding strains obtained for the ROM and diagnostic testing. All strains during ROM and treatment testing were dramatically smaller than the initial failure strains of the ICA. CONCLUSIONS: This study showed that maximal ICA strains imparted by cervical spinal manipulative treatments were well within the normal ROM. Chiropractic manipulation of the neck did not cause strains to the ICA in excess of those experienced during normal everyday movements. Therefore, cervical spinal manipulative therapy as performed by the trained clinicians in this study, did not appear to place undue strain on the ICA and thus does not seem to be a factor in ICA injuries.

Titin force is enhanced in actively stretched skeletal muscle.

The sliding filament theory of muscle contraction is widely accepted as the means by which muscles generate force during activation. Within the constraints of this theory, isometric, steady-state force produced during muscle activation is proportional to the amount of filament overlap. Previous studies from our laboratory demonstrated enhanced titin-based force in myofibrils that were actively stretched to lengths which exceeded filament overlap. This observation cannot be explained by the sliding filament theory. The aim of the present study was to further investigate the enhanced state of titin during active stretch. Specifically, we confirm that this enhanced state of force is observed in a mouse model and quantify the contribution of calcium to this force. Titin-based force was increased by up to four times that of passive force during active stretch of isolated myofibrils. Enhanced titin-based force has now been demonstrated in two distinct animal models, suggesting that modulation of titin-based force during active stretch is an inherent property of skeletal muscle. Our results also demonstrated that 15% of the enhanced state of titin can be attributed to direct calcium effects on the protein, presumably a stiffening of the protein upon calcium binding to the E-rich region of the PEVK segment and selected Ig domain segments. We suggest that the remaining unexplained 85% of this extra force results from titin binding to the thin filament. With this enhanced force confirmed in the mouse model, future studies will aim to elucidate the proposed titin-thin filament interaction in actively stretched sarcomeres.

Alterations in patellofemoral kinematics following vastus medialis transection in the anterior cruciate ligament deficient rabbit knee.

BACKGROUND: Anterior cruciate ligament deficiency and quadriceps muscle weakness are considered to be important risk factors for aberrant patellar tracking and subsequent patellofemoral osteoarthritis. However, data from in vivo experiments looking at dynamic patellar joint kinematics and muscle force are scarce. Therefore, the purpose of this study was to evaluate the effects of anterior cruciate ligament transection and loss of vastus medialis force on patellar tracking in the rabbit knee in vivo. METHODS: Eight skeletally mature New Zealand White Rabbits, weighing 6.0kg (0.6kg standard deviation) were used. The experimental trials consisted of active, concentric and eccentric movements of the knee joint. Measurements were performed with the intact, the anterior cruciate ligament deficient, and the vastus medialis transected knee. Patellofemoral kinematics (shift, rotation) were quantified from high speed video. FINDINGS: Following anterior cruciate ligament transection, patellar tracking occurred more laterally, and caused a significant lateral rotation of the patella. The addition of vastus medialis transection did not alter patellar tracking or rotation significantly for any of the force-matched experimental conditions. INTERPRETATION: The loss of the anterior cruciate ligament results in lateral patellar shift and rotation while the loss of vastus medialis muscle force does not affect patellar tracking or rotation in the anterior cruciate ligament deficient knee. We suggest that the current results should be considered carefully in future interpretations of knee extensor imbalance. More research is needed to describe the contribution of vastus medialis muscle strength to medial patellofemoral stability and confirm these results in the human knee.

Dynamic in-vivo force transfer in the lapine knee loaded by quadriceps muscle contraction.

BACKGROUND: The rabbit knee is a frequently used model for experimental osteoarthritis (OA). Despite the acknowledged importance of joint loading in the onset and progression of OA, the load transfer in the three compartments of the intact rabbit knee remains unknown. Therefore, this study was aimed at determining load transfer in the three compartments for isometric, concentric, and eccentric knee extensor contractions. METHODS: Maximal and sub-maximal isometric, concentric, and eccentric knee extensor contractions were produced by electrical stimulation of the femoral nerve in 13 rabbits. Knee extensor forces were measured using a custom-built servomotor. Contact areas and pressure distributions were measured in the patello-femoral, and the medial and lateral tibio-femoral joints using Fuji Presensor film. FINDINGS: Contact areas and peak pressures increased with increasing quadriceps forces for all compartments. Maximal knee extensor forces, joint moments, and contact pressures reached values of 504 N, 5.5 Nm and 60 MPa, respectively. Force transfer in the patello-femoral joint was about twice that observed in the individual tibio-femoral joints. During isometric contractions, force transfer was higher in the medial compared to the lateral tibio-femoral joint, while this trend was reversed for dynamic contractions. INTERPRETATION: The results of this study suggest that the increasing muscular forces are transferred through an increased contact area, thereby limiting the increase in average contact pressure. These results may be used as reference data for contact pressures in the intact rabbit knee and may form the foundation for studies using the lapine knee as an experimental model of osteoarthritis.

Are titin properties reflected in single myofibrils?

Titin is a structural protein in muscle that spans the half sarcomere from Z-band to M-line. Although there are selected studies on titin's mechanical properties from tests on isolated molecules or titin fragments, little is known about its behavior within the structural confines of a sarcomere. Here, we tested the hypothesis that titin properties might be reflected well in single myofibrils. Single myofibrils from rabbit psoas were prepared for measurement of passive stretch-shortening cycles at lengths where passive titin forces occur. Three repeat stretch-shortening cycles with magnitudes between 1.0 and 3.0µm/sarcomere were performed at a speed of 0.1µm/s·sarcomere and repeated after a ten minute rest at zero force. These tests were performed in a relaxation solution (passive) and an activation solution (active) where cross-bridge attachment was inhibited with 2,3 butanedionemonoxime. Myofibrils behaved viscoelastically producing an increased efficiency with repeat stretch-shortening cycles, but a decreased efficiency with increasing stretch magnitudes. Furthermore, we observed a first distinct inflection point in the force-elongation curve at an average sarcomere length of 3.5µm that was associated with an average force of 68±5nN/mm. This inflection point was thought to reflect the onset of Ig domain unfolding and was missing after a ten minute rest at zero force, suggesting a lack of spontaneous Ig domain refolding. These passive myofibrillar properties observed here are consistent with those observed in isolated titin molecules, suggesting that the mechanics of titin are well preserved in isolated myofibrils, and thus, can be studied readily in myofibrils, rather than in the extremely difficult and labile single titin preparations.

The three filament model of skeletal muscle stability and force production.

Ever since the 1950s, muscle force regulation has been associated with the cross-bridge interactions between the two contractile filaments, actin and myosin. This gave rise to what is referred to as the "two-filament sarcomere model". This model does not predict eccentric muscle contractions well, produces instability of myosin alignment and force production on the descending limb of the force-length relationship, and cannot account for the vastly decreased ATP requirements of actively stretched muscles. Over the past decade, we and others, identified that a third myofilament, titin, plays an important role in stabilizing the sarcomere and the myosin filament. Here, we demonstrate additionally how titin is an active participant in muscle force regulation by changing its stiffness in an activation/force dependent manner and by binding to actin, thereby adjusting its free spring length. Therefore, we propose that skeletal muscle force regulation is based on a three filament model that includes titin, rather than a two filament model consisting only of actin and myosin filaments.

The force-length relationship of the cat soleus muscle.

According to the force-length relationship, cat soleus optimal sarcomere length should lie between 2.3-2.5µm. Rack and Westbury (1969) found optimal sarcomere length around 2.8- 3.0µm. The purpose of this study was to repeat their study to check for these discrepancies between the expected and the measured optimal length. The soleus muscle of both hindlimbs of three cats was supra-maximally stimulated. Isometric forces were measured for lengths ranging from -20 to +20mm relative to the optimal length. Mean sarcomere lengths were obtained by laser diffraction. Fibre length was obtained post-mortem by video analysis and in situ with sonomicrometry crystals. Sarcomere number was determined and in situ sarcomere lengths were calculated. The sarcomere force-length relationship showed an ascending and descending part with a plateau between 2.0-2.4µm. Peak forces were obtained at smaller average sarcomere lengths than reported by Rack and Westbury and closer to the optimal sarcomere length based on sliding filament considerations.

A comparison of stage of presentation for pancreatic and colorectal cancer in Pennsylvania 2000-2005.

BACKGROUND: The goal of this study was to examine how rurality, socioeconomic status (SES) and access to medical care are related to the stage at presentation of patients with colorectal (CRC) and pancreatic cancer (PC) in Pennsylvania. MATERIALS AND METHODS: Incident CRC and PC cases were identified from the Pennsylvania Department of Health. Demographic, SES, and access variables were collected at the county level. RESULTS: Increased urbanization, younger age, and male gender were shown to be significantly related to later stage at diagnosis for PC. Age and education level were significant predictors of the rate of PC, while age, education level, insurance status, rurality, and the ratio of oncologists to primary care physicians were significant predictors of the rate of CRC. CONCLUSION: Based on county-level data, urban residence, younger age, and male gender were shown to be predictors of later stage at diagnosis for PC. These findings should help guide further research into factors that may be important predictors of later stage of diagnosis.

Does residual force enhancement increase with increasing stretch magnitudes?

It is generally accepted that force enhancement in skeletal muscles increases with increasing stretch magnitudes. However, this property has not been tested across supra-physiological stretch magnitudes and different muscle lengths, thus it is not known whether this is a generic property of skeletal muscle, or merely a property that holds for small stretch magnitudes within the physiological range. Six cat soleus muscles were actively stretched with magnitudes varying from 3 to 24 mm at three different parts of the force-length relationship to test the hypothesis that force enhancement increases with increasing stretch magnitude, independent of muscle length. Residual force enhancement increased consistently with stretch amplitudes on the descending limb of the force-length relationship up to a threshold value, after which it reached a plateau. Force enhancement did not increase with stretch amplitude on the ascending limb of the force-length relationship. Passive force enhancement was observed for all test conditions, and paralleled the behavior of the residual force enhancement. Force enhancement increased with stretch magnitude when stretching occurred at lengths where there was natural passive force within the muscle. These results suggest that force enhancement does not increase unconditionally with increasing stretch magnitude, as is generally accepted, and that increasing force enhancement with stretch appears to be tightly linked to that part of the force-length relationship where there is naturally occurring passive force.

Lymphoepithelioma-like carcinoma of the ureter discovered intraoperatively during a hysterectomy.

We present a patient with a T2NXMX lymphoepithelioma-like carcinoma (LELC) of the lower third of her left ureter discovered incidentally during removal of a large uterine mass. This case of LELC is unique for its presentation in the context of fibroid mass and its distinct (incidental) manner of discovery. To our knowledge, this will be the sixth case report to describe LELC of the ureter. A review of available literature and summary of upper tract cases are provided.

Premature deactivation of soleus during the propulsive phase of cat jumping.

It has been shown that cat soleus (SOL) forces remain nearly constant despite increases in electromyography (EMG) activity for increasing speeds of locomotion, while medial gastrocnemius (MG) forces and EMG activity increase in parallel. Furthermore, during jumping, average cat SOL forces decrease, while average EMG activity increases dramatically compared with walking conditions. Finally, during rapid paw-shake movements, SOL forces and EMG activities are nearly zero. Based on these results, we hypothesized that the SOL is deactivated, despite ankle extensor requirements, if the contractile conditions limit SOL force potential severely. The purposes of this study were to (i) investigate SOL EMG activity and force as a function of its contractile conditions during jumping, (ii) test whether SOL EMG activity is associated with SOL contractile conditions, and (iii) determine the functional implications of SOL EMG activity during jumping. It was found that the SOL was prematurely deactivated in two distinct phases during the propulsive phase of jumping, in which shortening speeds approached or even exceeded the maximal speed of muscle shortening. We concluded that the SOL was prematurely deactivated to save energy because its mechanical work output approached zero, and speculated that the first phase of deactivation might be caused by a decrease in group Ia firing associated with active shortening and the second by a pre-programmed response inherent to the central pattern generator.

History-dependence of isometric muscle force: effect of prior stretch or shortening amplitude.

It is well-recognised that steady-state isometric muscle force is decreased following active shortening (force depression, FD) and increased following active stretch (force enhancement, FE). It has also been demonstrated that passive muscle force is increased following active stretch (passive FE). Several studies have reported that FD increases with shortening amplitude and that FE and passive FE increase with stretch amplitude. Here, we investigate whether these trends continue with further increases in shortening or stretch amplitude. Experiments were performed using in situ cat soleus muscles (n=8 for FD; n=7 for FE and passive FE). FD, FE and passive FE were measured after shortening or stretch contractions that covered as wide a range of amplitudes as practically possible without damaging the muscles. FD increased approximately linearly with shortening amplitude, over the full range of amplitudes investigated. This is consistent with the hypothesis that FD arises from a stress-induced inhibition of crossbridges. FE increased with stretch amplitude only up to a point, and then levelled off. Passive FE, and the transient increase in force at the end of stretch, showed relationships to stretch amplitude that were qualitatively very similar to the relationship for FE, increasing only until the same critical stretch amplitude had been reached. We conclude that FE and passive FE do not increase with stretch amplitude under all circumstances. This finding has important consequences for determining the mechanisms underlying FE and passive FE because any mechanism that is proposed to explain them must be able to predict it.

Control of ground reaction forces by hindlimb muscles during cat locomotion.

It has been proposed that biarticular muscles are primarily responsible for the control of the direction of external forces, as their activation is closely related and highly sensitive to the direction of external forces. This functional role for biarticular muscles has been supported qualitatively by experimental evidence, but has never been tested quantitatively for lack of a mathematical/mechanical formulation of this theory and the difficulty of measuring individual muscle forces during voluntary movements. The purposes of this study were: (1) to define rules for muscular coordination based on the control of external forces; (2) to develop a model of the cat hindlimb that allows for the calculation of the magnitude and direction of the ground reaction forces (GRFs) produced by individual hindlimb muscles; and (3) to test if the coordination of mono- and biarticular cat hindlimb muscles is related to the control of the resultant GRF. We measured the GRF, hindlimb kinematics, selected muscle forces and activations during cat locomotion. Then, the measured muscle forces were used as input to the hindlimb model to compute the muscle-induced GRF. We assume that if activation (and possibly force) increased as the muscle-induced component of GRF approximated the resultant GRF, then that muscle was used by the central nervous system (CNS) to help control the direction of the external GRF. During cat walking, medial gastrocnemius (MG) and plantaris (PL) forces increased with increasing proximity to the GRF, while soleus (SOL) forces and vastus lateralis (VL) activations did not. SOL and VL activation were most strongly related to the vertical and parallel (braking/accelerating) component of the GRF, respectively. We concluded from these results that MG and PL are primarily responsible for the control of the direction of the GRF, while SOL primarily functions as an anti-gravity muscle, and VL as an acceleration/deceleration muscle.

Multi-functionality of the cat medical gastrocnemius during locomotion.

The functional role of biarticular muscles was investigated based on direct force measurement in the cat medial gastrocnemius (MG) and analysis of hindlimb kinematics and kinetics for the stance phase of level, uphill, and downhill walking. Four primary functional roles of biarticular muscles have been proposed in the past. These functional roles have typically been discussed independently of each other, and biarticular muscles have rarely been assigned more than one functional roles for different phases of the work cycle. The purpose of this study was to elucidate the functional role of the biarticular cat MG during locomotion. It was found that MG forces were primarily associated with the moment requirements at the ankle for most of the stance phase, but also helped to satisfy the moments at the knee in the initial phase of stance. In the second half of stance, MG transferred mechanical energy from the knee to the ankle from the knee to the ankle, while simultaneously producing a substantial amount of mechanical work. Based on these results, we hypothesize that MG's primary function is that of an ankle extensor. However, because of the coupling of the ankle extensor moment with a knee flexor moment in the initial, and a knee extensor moment in the final phase of stance, MG satisfies two joint moments in early stance, and transfers mechanical energy from the knee to the ankle in late stance. We conclude that cat MG has multiple functional roles during the stance phase of locomotion, and speculate that such multi-functionality also exists in other bi- and multi-articular muscles.

Coordination of medial gastrocnemius and soleus forces during cat locomotion.

We studied force-sharing behavior between the cat medial gastrocnemius (MG) and soleus (SOL) muscles by direct measurement of the muscle forces and electromyographic activities (EMGs), muscle lengths, speeds of contraction, joint kinematics and kinetics, for a variety of locomotor conditions. Previous studies suggested that the modulation of MG force and activation is associated with movement demands, while SOL force and activation remain nearly constant. However, no systematic, quantitative analysis has been done to evaluate the degree of (possible) modulation of SOL force and activation across a range of vastly different locomotor conditions. In the present study, we investigated the effects of speed and intensity of locomotion on the modulation of SOL force and EMG activity, based on quantitative, statistical analyses. We also investigated the hypothesis that MG forces are primarily associated with MG activation for changing movement demands, while SOL forces are primarily associated with the contractile conditions, rather than activation. Seven cats were trained to walk, trot and gallop at different speeds on a motor-driven treadmill, and to walk up and down different slopes on a walkway. Statistical analysis suggested that SOL activation (EMG activity) significantly increased with increasing speeds and intensities of locomotion, while SOL forces remained constant in these situations. MG forces and EMG activities, however, both increased with increasing speeds and intensities of locomotion. We conclude from these results that SOL is not maximally activated at slow walking, as suggested in the literature, and that its force remains nearly constant for a range of locomotor conditions despite changes in EMG activity. Therefore, SOL forces appear to be affected substantially by the changing contractile conditions associated with changing movement demands. In contrast, MG peak forces correlated well with EMG activities, suggesting that MG forces are primarily associated with activation while its contractile conditions play a minor role for the movement conditions tested here.