ABNORMAL PASSIVE FORCES IN FROG TIBIALIS ANTERIOR MUSCLES

INTRODUCTION Typically in force-length relationships of skeletal muscle, the passive forces are much lower than the active forces. These results have been shown in both single fiber experiments and whole muscle experiments [1]. However, abnormally high passive forces were noticed in a pilot study done with the whole tibialis anterior (TA) in the frog, rana pipiens. Surprisingly, single fiber experiments of the same muscle and species have shown a standard force-length relationship with low passive force. Thus, the purpose of this study was to verify the observation of extremely high passive forces made previously and, if true, investigate the range of muscle lengths in which these high pasive forces occur. Possible functional reasons for this phenomenon will be discussed. METHODS The TA and sciatic nerve of three frogs were isolated with surgical techniques. The attached tarsal bone was clamped to a load cell and the knee was pinned. The muscle was then stretched to various lengths and isometrically contracted via nerve stimulation with a platinum hook. A Short rest was given between trials. Whole muscle biopsies were taken and prepared for analysis of sarcomere length within the normal range of motion. The range of muscle lengths in which the animal normally functions was analysed by configuring knee and ankle joint angles to values that were identified for swimming and jumping. [2]. RESULTS All subjects showed very large passive forces on the ascending limb of the force-length relationship (Figure 1). At long muscle lengths, passive forces were up to six times as high as the corresponding active forces. The functional range of muscle lengths for jumping and swimming is 87-95% and 91%-96%, respectivelyof the maximum in vivo muscle length. The shortest in vivo muscle length is about 77% of its maximal in vivo length. DISCUSSION AND CONCLUSIONS It is likely that the high passive forces observed in the frog TA come from the extracellular matrix, as earlier studies with single TA fibers show low passive force [3]. These high passive forces found for the entire muscle have implications for the function of the muscle during every day activities, such as swimming and jumping.   It has been demonstrated that frogs produce extremely high muscle power output during jumping [4]. They achieve these high powers by using a catapult mechanism through the plantaris tendon [5]. High passive forces in the TA would be a natural development to counterbalance plantaris forces as the TA is the antagonist muscle to the plantaris.   It is also possible that the high passive forces in the TA are used during frog swimming. Since frogs have large webbed feet, a high passive force in the TA allows frogs to use their feet like human swimmers use passive fins at to increase the speed and decrease the energy expenditure of swimming

Vertebral arteries do not experience tensile force during manual cervical spine manipulation applied to human cadavers

Background: The vertebral artery (VA) may be stretched and subsequently damaged during manual cervical spine manipulation. The objective of this study was to measure VA length changes that occur during cervical spine manipulation and to compare these to the VA failure length. Methods: Piezoelectric ultrasound crystals were implanted along the length of the VA (C1 to C7) and were used to measure length changes during cervical spine manipulation of seven un-embalmed, post-rigor human cadavers. Arteries were then excised, and elongation from arbitrary in-situ head/neck positions to first force (0.1 N) was measured. Following this, VA were stretched (8.33 mm/s) to mechanical failure. Failure was defined as the instance when VA elongation resulted in a decrease in force. Results: From arbitrary in-situ head/neck positions, the greatest average VA length change during spinal manipulation was [mean (range)] 5.1% (1.1 to 15.1%). From arbitrary in-situ head/neck positions, arteries were elongated on average 33.5% (4.6 to 84.6%) prior to first force occurrence and 51.3% (16.3 to 105.1%) to failure. Average failure forces were 3.4 N (1.4 to 9.7 N). Conclusions: Measured in arbitrary in-situ head/neck positions, VA were slack. It appears that this slack must be taken up prior to VA experiencing tensile force. During cervical spine manipulations (using cervical spine extension and rotation), arterial length changes remained below that slack length, suggesting that VA elongated but were not stretched during the manipulation. However, in order to answer the question if cervical spine manipulation is safe from a mechanical perspective, the testing performed here needs to be repeated using a defined in-situ head/neck position and take into consideration other structures (e.g. carotid arteries). Keywords: Spinal biomechanics; cerebrovascular accidents; spinal manipulation; stroke; vertebral artery dissection

DIET INDUCED OBESITY MAY AFFECT THE FORCE-VELOCITY RELATIONSHIP IN RAT SOLEUS

INTRODUCTION Obesity is associated with chronic, low-grade inflammation that has been shown to affect several musculoskeletal tissues [1].  Previously, we observed that diet induced obesity (DIO) using a high-fat, high sugar diet results in molecular and morphological alterations in muscles, including an increase in intramuscular fat. However, it remains unclear if these alterations affect muscle function, as few studies have characterized the functional properties of muscles in obese individuals [2]. Arguably, one of the most important functional properties of skeletal muscle is the force-velocity relationship (FVR).  When muscles are shortening at increasing velocities, force decays exponentially as the proportion of attached cross bridges and the average force per cross-bridge decreases. As fat infiltration reduces the contractile material and structure of obese muscles, the functional properties of muscles, specifically the FVR may be affected. Therefore, the purpose of this study was to characterize the FVR in rat soleus muscles obtained from obese and normal rats. We hypothesized that soleus muscles of obese rats would produce lower absolute and relative forces at any given shortening velocity compared with muscles from control animals. METHODS Outbred, individually housed male Sprague-Dawley rats, aged 10-12 weeks, were randomized to a high fat, high sugar diet (DIO, 40% fat 45% sucrose, n=9) or a standard chow diet (n=5, chow, 12% fat, 0% sucrose) for 12 weeks. Prior to surgery, animals were sedated, weighed, and body composition was quantified using dual energy X-ray absorptiometry. The right soleus muscle was exposed and a custom cuff-type electrode was implanted on the tibial nerve. The soleus tendon was isolated from the Achilles with the calcaneus attached and fixed to a motor along the muscle’s natural alignment. The muscle was then stretched to its optimum length and electrically stimulated at 35Hz at 2.5x the motor unit threshold [3]. An isometric force reading was acquired over 2.5 s stimulation. The soleus was then stretched past its optimum length and shortened at increasing velocities, a force reading was collected at optimal length using Windaq Software. Data were collected at 1000Hz. Forces during shortening were measured at the same length and time point as the isometric reference force. This process was repeated for shortening velocities increasing from 2 mm/s to 70 mm/s. Animals were sacrificed and soleus muscles were harvested and weighed. Data were processed using a custom Matlab ® zero-phase filter program. Instantaneous forces were normalized to the peak isometric force for each animal. Comparisons were made using a Student’s t-tests, α=0.05. RESULTS On average, DIO animals had higher body mass and body fat compared to the control rats (p<0.001). Soleus mass was similar between DIO animals and chow (p=0.321), as was peak active isometric force (DIO: 2.48±0.10 N, chow: 2.08±0.33 p=0.183). FVR relationships were statistically different at shortening velocities between 3 and 35 mm/s (DIO > Chow at each given velocity; p<0.05, Fig. 1). DISCUSSION The results opposed the expected outcomes of this study and, therefore, the hypothesis was not satisfied. These findings could be due to a higher proportion of fast twitch fibers in the DIO or longer fascicle lengths in the DIO rats, but these speculations remain to be tested. Another possible explanation involves a potential increase in sensitivity to stimulation in the DIO soleus muscle resulting in increased force. Future work will examine other structural levels and muscle contractile proteins to understand these preliminary findings

FUNCTIONAL EFFECTS OF DIET INDUCED OBESITY ON PERMEABLIZED RAT MUSCLE FIBRES

INTRODUCTION Muscle performance is determined by the metabolic, calcium handling, and sarcomeric characteristics of its constituent fibres. Diet induced obesity (DIO) may influence contractile performance in whole muscle, but little is known about the effects of DIO at the single fibre level. Particularly, how DIO might influence the contractile characteristics of single fibres free from the influence of metabolic or calcium handling properties is not established. There is some limited evidence to suggest that DIO may influence the sarcomeric proteins. For example, troponin T, an important regulatory protein found on the thin filament, exhibits a shift from the fast T3 isoform, to the slow T1 isoform, in mouse soleus muscle following high fat feeding, with no associated change in myosin heavy chain isoform [1]. These results suggest that DIO may cause fast fibres to express slow isoforms of sarcomeric proteins in postural muscles of mixed fibre-type. The purpose of this study was to assess the force-calcium and force-velocity relationships of skinned fast and slow fibres of vastus intermedius, a mixed fibre-type postural muscle [2], in chow-fed rats and a rat model of DIO. It was hypothesized that fast fibres from DIO rats would exhibit characteristics associated with a slower fibre phenotype, including increased calcium sensitivity and lower shortening velocities. METHODS Individually housed male Sprague-Dawley rats, aged 10-12 weeks, were randomized to undergo diet induced obesity (DIO) where they were fed a high fat, high sugar diet (n = 6) or a standard chow diet (n = 6) for 12 weeks. The caloric content of DIO diets was 40% fat and 45% sucrose, compared to chow diets which consisted of 12% fat and 0% sucrose. Both vastus intermedius muscles were collected from each rat. Similar to the mouse soleus, the fibre type distribution of rat vastus intermedius is approximately 50% type I and 50% type IIa fibres [2]. Muscles were chemically skinned in a glycerol-rigor solution for 2 weeks.  Two fast and two slow fibres per animal were then isolated and mounted in a model 802B skinned fibre test system (Aurora Scientific) at 2.4 µm sarcomere length for testing.  Preliminary fibre type assessment was made using a strontium sensitivity test [3].  The force-pCa relationship was assessed from pCa 7.2 to pCa 4.2. The force-velocity relationship was assessed by measuring the shortening velocity during isotonic contractions.  Maximal shortening velocity (Vmax) was assessed by a slack test protocol. Statistical differences were determined using Student’s t-test or a two-way factorial ANOVA and Newman-Keuls post-hoc analysis as appropriate, α = 0.05. RESULTS Dual-energy X-ray absorptiometry scans revealed DIO rats had significantly higher body mass, fat mass, and greater percent body fat than chow fed rats (all p<0.05), while lean mass was not significantly different between groups. DIO did not affect the force per cross-sectional area (CSA) of skinned fibres (Table 1). Fast DIO fibres had significantly lower maximum shortening velocities when compared to fast chow fibres (p<0.05; Table 1). No such differences were observed in slow fibres. Independent of fibre type, DIO fibres had significantly higher calcium sensitivity than chow fibres (p<0.01, Table 1).  While the Hill coefficient of the force pCa relationship was different between fast and slow chow fibres, no differences were seen in DIO fibres (p<0.05; Table 1).DISCUSSION AND CONCLUSIONS Consistent with a fast to slow phenotype transition, Vmax was lower in fast DIO fibres. However, DIO influenced the force-calcium relationship of both fast and slow fibres. Therefore, adaptations are not limited to fast fibres, but rather influence contractility on a larger scale. Whether this influence is global or localized to postural muscles remains to be determined. The specific isoforms of contractile proteins expressed in single fibres should be assessed following DIO

THE EFFECTS OF DIET INDUCED OBESITY ON THE FORCE-LENGTH RELATIONSHIP IN RAT SOLEUS

INTRODUCTION Obesity is associated with chronic inflammation, which has been shown to affect the integrity of musculoskeletal tissues [1]. Previous data from our group suggests that obesity can result in intramuscular fat deposition [1]. It is unclear if this structural alteration has functional consequences, as the implications of obesity on muscle mechanics are not well understood. Therefore, the purpose of this study was to quantify the active force produced by soleus muscles of obese and non-obese rats at a range of muscle lengths. As the inclusion of fat into the muscle fibers will leave less room for contractile proteins, we hypothesized that obese rats will produce lower forces normalized to muscle mass at every length than non-obese control rats.   METHODS Fourteen rats were randomly allocated to a 12-week diet: either an obesity-inducing high fat high sucrose diet (DIO, 40% fat, 45% sucrose, n=8) or a standard chow diet (chow, 12% fat 0% sucrose, n=6). Prior to surgery, body composition was evaluated using dual energy X-ray absorptiometry. Custom-made tibial nerve cuffs were surgically attached to the right tibial nerve of each animal. The soleus was exposed, mechanically isolated, and clamped to a force transducer. The muscle was then stretched to a predetermined length and electrically stimulated at 3 times the motor unit threshold (50Hz) and the force output was measured [3]. Force tracings were digitized using WINDAQ® software. Passive, active, and total forces produced by the soleus were normalized to the maximum in vivo length of each animal. Forces were averaged into 5% length intervals within each animal. Students t-tests or a two-way ANOVA were conducted between groups, and a Bonferroni correction was used as needed, α=0.05. RESULTS DIO rats had increased body mass (DIO 816.4 ± 30.1g, chow 645.0 ± 28.3g; p<0.05) and body fat (DIO 39.2 ± 1.3%, chow 21.8 ± 2.1%; p<0.05) compared to chow-fed rats. Soleus mass (DIO: 0.28 ± 0.01 g, chow: 0.26 ± 0.11 g, p=0.32), was similar between the two groups. Absolute peak isometric force was similar between the two groups (DIO: 2.58 ± 0.10 N, chow: 2.18 ± 0.34 N, p=0.23). Active isometric force normalized to soleus mass was significantly higher in DIO group rats at every muscle length (Figure 1, p<0.05). DISCUSSION AND CONCLUSIONS On average, DIO rats produced more active force at a given normalized length and soleus mass than chow rats, a finding that refutes our original hypothesis. Since optimal length occurs at the same relative muscle length for both groups, and since the decline in force from maximum is similar between groups, it appears that fascicle length, and an associated shift in the force-length relationship cannot explain our results. Results of differences in the force-velocity relationship (not shown here) suggest that the DIO rats may have a higher proportion of fast twitch fibres, but the relative force among slow and fast fibres is similar, and thus also should not affect these results. The results suggest that the force per cross-sectional area is higher in muscles from obese compared to lean rats, a finding that defies explanation at this time and needs thorough investigation in the future. Histology and tests looking at fibre and cell level muscle structures may provide more insight

Anterior cruciate ligament transection alters the n-3/n-6 fatty acid balance in the lapine infrapatellar fat pad

The infrapatellar fat pad (IFP) of the knee joint has received lots of attention recently due to its emerging role in the pathogenesis of osteoarthritis (OA), where it displays an inflammatory phenotype. The aim of the present study was to examine the infrapatellar fatty acid (FA) composition in a rabbit (Oryctolagus cuniculus) model of early OA created by anterior cruciate ligament transection (ACLT).Peer reviewe

The Eyes Have It: Sex and Sexual Orientation Differences in Pupil Dilation Patterns

Recent research suggests profound sex and sexual orientation differences in sexual response. These results, however, are based on measures of genital arousal, which have potential limitations such as volunteer bias and differential measures for the sexes. The present study introduces a measure less affected by these limitations. We assessed the pupil dilation of 325 men and women of various sexual orientations to male and female erotic stimuli. Results supported hypotheses. In general, self-reported sexual orientation corresponded with pupil dilation to men and women. Among men, substantial dilation to both sexes was most common in bisexual-identified men. In contrast, among women, substantial dilation to both sexes was most common in heterosexual-identified women. Possible reasons for these differences are discussed. Because the measure of pupil dilation is less invasive than previous measures of sexual response, it allows for studying diverse age and cultural populations, usually not included in sexuality research

The Effect of Vastus Medialis Transection on Patellofemoral Contact Pressure and Patellar Tracking

The purpose of this thesis was to examine the effects of imbalance between the forces exerted by vastus medialis (VM) and vastus lateralis on patellofemoral peak contact pressures, areas, shapes, and patellar tracking before and after the removal of VM. New Zealand White rabbits were used and patellofemoral contact mechanics were evaluated at 30°, 60° and 90° and patellar tracking was recorded from 30°-90° before and after VM transection. Following removal of VM, there were no changes in patellofemoral contact mechanics and patellar tracking. We conclude that VM weakness does not cause changes in rabbit patellofemoral contact mechanics. Since muscular alignment and knee joint geometry are similar in human and rabbits, we question the idea of VM weakness as a cause for patellar mal-tracking and patellofemoral joint pain

Effect of Vastus Medialis Loss on Rabbit Patellofemoral Joint Contact Pressure Distribution

Vastus medialis (VM) weakness is thought to alter patellar tracking, thereby changing the loading of the patellofemoral joint (PFJ), resulting in patellofemoral pain. However, it is challenging to measure VM force and weakness in human studies, nor is it possible to measure the associated mechanical changes in the PFJ. To obtain fundamental insight into VM weakness and its effects on PFJ mechanics, the authors determined PFJ loading in the presence of experimentally simulated VM weakness. Skeletally mature New Zealand White rabbits were used (n = 6), and the vastus lateralis, VM, and rectus femoris were stimulated individually through 3 custom-built nerve cuff electrodes. Muscle torque and PFJ pressure distribution were measured while activating all muscles simultaneously, or when the vastus lateralis and rectus femoris were activated, while VM was not, to simulate a quadriceps muscle strength imbalance. For a given muscular joint torque, peak pressures were greater and joint contact areas were smaller when simulating VM weakness compared with the condition where all muscles were activated simultaneously. The results in the rabbit model support that VM weakness results in altered PFJ loading, which may cause patellofemoral pain, often associated with a strength imbalance of the knee extensor muscle group.Canadian Institutes of Health Research (CIHR)Othe

Contribution of individual quadriceps muscles to knee joint mechanics

Many attempts have been made to determine the contribution of individual muscles in an agonistic group to the mechanics of joints. However, previous approaches had the limitations that muscles often could not be controlled in a precise manner, that individual muscles in an agonistic group could not be activated individually, and that individual muscle contributions could not be measured in an actively contracting agonistic group. Here, we introduce a surgical approach that allows for controlled activation of individual muscles of an agonistic group. The approach is illustrated for the vastus lateralis (VL), vastus medialis (VM) and rectus femoris (RF) of the rabbit quadriceps femoris group. We provide exemplar results for potential applications of the approach, such as measuring the pressure distribution in the patellofemoral joint, and the torque–angle relationship of VL, VM and RF when activated individually and when the three muscles are activated simultaneously.Natural Sciences and Engineering Research Council - Collaborative Research & Development GrantUniversity of Calgary - Research GrantOthe