Passive Tension Research Articles

Changes in Muscle Stress and Sarcomere Adaptation in Mice Following Ischemic Stroke.

While abnormal muscle tone has been observed in people with stroke, how these changes in muscle tension affect sarcomere morphology remains unclear. The purpose of this study was to examine time-course changes in passive muscle fiber tension and sarcomeric adaptation to these changes post-ischemic stroke in a mouse model by using a novel in-vivo force microscope. Twenty-one mice were evenly divided into three groups based on the time point of testing: 3 days (D3), 10 days (D10), and 20 days (D20) following right middle cerebral artery ligation. At each testing time, the muscle length, width, and estimated volume of the isolated soleus muscle were recorded, subsequently followed by in-vivo muscle tension and sarcomere length measurement. The mass of the soleus muscle was measured at the end of testing to calculate muscle density. Two-way ANOVA with repeated measures was used to examine the differences in each of the dependent variable among the three time-point groups and between the two legs. The passive muscle stress of the impaired limbs in the D3 group (27.65 ± 8.37 kPa) was significantly lower than the less involved limbs (42.03 ± 18.61 kPa; p = 0.05) and the impaired limbs of the D10 (48.92 ± 14.73; p = 0.03) and D20 (53.28 ± 20.54 kPa; p = 0.01) groups. The soleus muscle density of the impaired limbs in the D3 group (0.69 ± 0.12 g/cm3) was significantly lower than the less involved limbs (0.80 ± 0.09 g/cm3; p = 0.04) and the impaired limbs of the D10 (0.87 ± 0.12 g/cm3; p = 0.02) and D20 (1.00 ± 0.14 g/cm3; p < 0.01) groups. The D3 group had a shorter sarcomere length (2.55 ± 0.26 μm) than the D10 (2.83 ± 0.20 μm; p = 0.03) and D20 group (2.81 ± 0.15 μm; p = 0.04). These results suggest that, while ischemic stroke may cause considerable changes in muscle tension and stress, sarcomere additions under increased mechanical loadings may be absent or disrupted post-stroke, which may contribute to muscle spasticity and/or joint contracture commonly observed in patients following stroke.

C-type natriuretic peptide moderates titin-based cardiomyocyte stiffness.

Heart failure is often accompanied by titin-dependent myocardial stiffness. Phosphorylation of titin by cGMP-dependent protein kinase I (PKGI) increases cardiomyocyte distensibility. The upstream pathways stimulating PKGI-mediated titin phosphorylation are unclear. We studied whether C-type natriuretic peptide (CNP), via its guanylyl cyclase-B (GC-B) receptor and cGMP/PKGI signaling, modulates titin-based ventricular compliance. To dissect GC-B–mediated effects of endogenous CNP in cardiomyocytes, we generated mice with cardiomyocyte-restricted GC-B deletion (CM GC-B–KO mice). The impact on heart morphology and function, myocyte passive tension, and titin isoform expression and phosphorylation was studied at baseline and after increased afterload induced by transverse aortic constriction (TAC). Pressure overload increased left ventricular endothelial CNP expression, with an early peak after 3 days. Concomitantly, titin phosphorylation at Ser4080, the site phosphorylated by PKGI, was augmented. Notably, in CM GC-B–KO mice this titin response was abolished. TAC-induced hypertrophy and fibrosis were not different between genotypes. However, the KO mice presented mild systolic and diastolic dysfunction together with myocyte stiffness, which were not observed in control littermates. In vitro, recombinant PKGI rescued reduced titin-Ser4080 phosphorylation and reverted passive stiffness of GC-B–deficient cardiomyocytes. CNP-induced activation of GC-B/cGMP/PKGI signaling in cardiomyocytes provides a protecting regulatory circuit preventing titin-based myocyte stiffening during early phases of pressure overload.

Abstract 13402: FHL-1 Contributes to and Colocalizes With Titin in Cardiac Hypertrophy

Increased myocardial stiffness is a hallmark of impaired diastole in heart failure (HF). Hypo-phosphorylation of the N2B unique sequence (N2Bus) of titin (TTN), a giant myofilament protein, increases passive tension leading to diastolic dysfunction in HF. Enhancing the altered N2Bus phosphorylation improves cardiac stiffness and function. FHL-1, an interacting protein potentially modulating N2Bus phosphorylation is increased in HF. FHL-1 knockout mice display blunted cardiac hypertrophy and improved diastolic compliance in response to pressure-overload by transverse aortic constriction, TAC. FHL-1 also regulates skeletal muscle hypertrophy. We hypothesize that FHL-1 contributes to cardiac hypertrophy and colocalizes/interacts with TTN in heart failure. In house IHC data showed site-specific N2Bus hypo-phosphorylation at S4099, S4010 and S4185 in human HCM and/or DCM tissue samples. Longitudinal in-vivo studies showed that FHL-1 and cardiac hypertrophy markers genes were increased in left ventricles (LV) of TAC mice using RNA-seq. A persistently enhanced FHL-1 protein expression by immunoblotting and mass spectrometry strongly correlated with LV hypertrophy at 1, 4, and 6 weeks post-TAC. In addition, LV hypertrophy correlated negatively with function (fractional shortening). Increases in FHL-1 and hypertrophy markers mRNA levels were confirmed by RT-qPCR in neonatal rat ventricular myocytes (NRVM) under phenylephrine (PE) and endothelin-1 (ET-1) induced hypertrophy. Under similar conditions, FHL-1 protein levels were increased by immunofluorescence (IF) in cytoplasmic, perinuclear and nuclear regions of NRVM. Colocalization of phospho-TTN and FHL-1 was observed in NRVM and was enhanced under PE- and ET-1 induced hypertrophy. IF studies in human skeletal myotubes showed that FHL-1 expression was increased during myoblast differentiation and IGF1-E3R induced hypertrophy. Preliminary data using Microscale Thermophoresis showed binding affinity between N2Bus and FHL-1 proteins. Our studies show that FHL-1 contributes to hypertrophy in addition to N2B hypo-phosphorylation status contributing to diastolic dysfunction in HF. Targeting FHL-1 and TTN can be a potential strategy to improve diastolic compliance in HF.

Os Acromiale in Reverse Total Shoulder Arthroplasty: A Cohort Study

Background:Increased passive deltoid tension after reverse total shoulder arthroplasty (RTSA) potentially leads to displacement or tilting of a preexisting os acromiale.Purpose:To analyze patients with an os acromiale who underwent RTSA and compare their outcomes and complications with a matched control group without an os acromiale.Study Design:Cohort study; Level of evidence, 3.Methods:In this study, 45 shoulders in 42 patients with an os acromiale (cases) were matched to 133 patients without os acromiale (controls) who underwent RTSA between 2005 and 2016. The mean follow-up was 52 ± 32 months. Matching criteria included sex, type of surgery, duration of follow-up, and age. The Constant score (CS), Subjective Shoulder Value (SSV), and radiological outcomes were assessed postoperatively at 1-year, 2-year, and final follow-up visits.Results:The mean CS, SSV, and range of motion improved from preoperative levels to the final follow-up in both groups (P < .01). Patients with an os acromiale had a relative CS of 70 ± 23 versus 76 ± 21 points (P = .15) and an SSV of 70 ± 30 versus 73 ± 24 (P = .52) compared with controls at the final follow-up visit. Patients with an os acromiale had significantly decreased active flexion of 104° ± 33° versus 114° ± 33° (P = .03) at 1 year and active abduction of 103° ± 37° versus 121° ± 38° at 2 years postoperatively (P = .02). A postoperatively painful os acromiale was found in 12 cases (27%) and spontaneously resolved in 8 cases after a mean of 33 months (range, 12-47 months; P = .04).Conclusion:RTSA reliably restores patient satisfaction despite the presence of an os acromiale, with a slightly impaired range of motion. Postoperative local tenderness at the os acromiale can be expected in 1 out of 4 patients, but this resolves spontaneously over time in the majority of patients.

In Vivo Pressurization of the Zebrafish Embryonic Heart as a Tool to Characterize Tissue Properties During Development.

Cardiac morphogenesis requires an intricate orchestration of mechanical stress to sculpt the heart as it transitions from a straight tube to a multichambered adult heart. Mechanical properties are fundamental to this process, involved in a complex interplay with function, morphology, and mechanotransduction. In the current work, we propose a pressurization technique applied to the zebrafish atrium to quantify mechanical properties of the myocardium under passive tension. By further measuring deformation, we obtain a pressure-stretch relationship that is used to identify constitutive models of the zebrafish embryonic cardiac tissue. Two-dimensional results are compared with a three-dimensional finite element analysis based on reconstructed embryonic heart geometry. Through these steps, we found that the myocardium of zebrafish results in a stiffness on the order of 10kPa immediately after the looping stage of development. This work enables the ability to determine how these properties change under normal and pathological heart development.

Effective stretching positions for the posterior shoulder capsule as determined by shear wave elastography

Stretching is often used to prevent and treat posterior shoulder capsule tightness; however, the most effective stretching positions are not clearly defined. The purpose of this study was to identify the stretching positions that specifically applied the greatest passive tension on the posterior shoulder capsule by evaluating the elastic characteristics of posterior capsules and muscles in various stretching positions using ultrasound shear wave elastography (SWE). We evaluated 9 fresh-frozen shoulders (mean age 86.6 ± 7.7 years) without osteoarthritis or rotator cuff tears. All posterior shoulder tissues were preserved intact. Shear moduli of the middle and inferior posterior shoulder capsules and the posterior shoulder muscles were evaluated using SWE. We obtained shear modulus measurements in 9 stretching positions using a combination of glenohumeral elevation planes and angles (frontal, sagittal, scapular; -30°, 0°, 30°, 60°, respectively). A 4-Nm torque for shoulder internal rotation or horizontal adduction was applied in each position. We also measured shear moduli in the resting position (0° elevation with neutral shoulder internal/external rotation). We compared the shear moduli of all stretching and resting positions using 1-way repeated measures analysis of variance (P < .05). In addition, we compared the shear modulus in 2 positions (ie, resting and each stretching) among tissues (ie, capsules and muscles) with repeated measures using 2-way analysis of variance (P < .05). Shear modulus values for the middle posterior capsules in "internal rotation at 30° in scapular plane elevation" (28.7 ± 14.3 kPa, P = .01) and in "horizontal adduction at 60° of elevation" (31.1 ± 13.1 kPa, P < .001) were significantly higher than that of the resting position (11.0 ± 7.3 kPa). The shear modulus value for the inferior posterior capsule in "internal rotation at 30° of flexion" was significantly higher than that of the resting position (39.0 ± 17.3 vs. 15.4 ± 13.9 kPa, respectively; P = .004). Additionally, the shear modulus values for the posterior capsules in "internal rotation at 30° in scapular plane elevation and flexion" were significantly higher than that of the posterior shoulder muscles. Effective middle posterior shoulder capsule stretching positions were shoulder "internal rotation at 30° of scapular plane elevation" and "horizontal adduction at 60° of elevation." Shoulder "internal rotation at 30° of flexion" was the most effective position for the inferior posterior shoulder capsule. Stretching in these positions could relieve posterior shoulder capsule tightness and contribute to the prevention and treatment of throwing injuries of the shoulder.

Myosin heavy chains in extraocular muscle fibres: Distribution, regulation and function.

This review examines kinetic properties and distribution of the 11 isoforms of myosin heavy chain (MyHC) expressed in extraocular muscle (EOM) fibre types and the regulation and function of these MyHCs. Although recruitment and discharge characteristics of ocular motoneurons during fixation and eye movements are well documented, work directly linking these properties with motor unit contractile speed and MyHC composition is lacking. Recruitment of motor units according to Henneman's size principle has some support in EOMs but needs consolidation. Both neurogenic and myogenic mechanisms regulate MyHC expression as in other muscle allotypes. Developmentally, multiply-innervated (MIFs) and singly-innervated fibres (SIFs) are derived presumably from distinct myoblast lineages, ending up expressing MyHCs in the slow and fast ends of the kinetic spectrum respectively. They modulate the synaptic inputs of their motoneurons through different retrogradely transported neurotrophins, thereby specifying their tonic and phasic impulse patterns. Immunohistochemical analyses of EOMs regenerating in situ and in limb muscle beds suggest that the very impulse patterns driving various ocular movements equip effectors with appropriate MyHC compositions and speeds to accomplish their tasks. These experiments also suggest that satellite cells of SIFs and MIFs are distinct lineages expressing different MyHCs during regeneration. MyHC compositions and functional characteristics of orbital fibres show longitudinal variations that facilitate linear ocular rotation during saccades. Palisade endings on global MIFs are postulated to respond to active and passive tensions by triggering axon reflexes that play important roles during fixation, saccades and vergence. How EOMs implement Listings law during ocular rotation is discussed.

Modifications of Titin Contribute to the Progression of Cardiomyopathy and Represent a Therapeutic Target for Treatment of Heart Failure

Titin is the largest human protein and an essential component of the cardiac sarcomere. With multiple immunoglobulin(Ig)-like domains that serve as molecular springs, titin contributes significantly to the passive tension, systolic function, and diastolic function of the heart. Mutations leading to early termination of titin are the most common genetic cause of dilated cardiomyopathy. Modifications of titin, which change protein length, and relative stiffness affect resting tension of the ventricle and are associated with acquired forms of heart failure. Transcriptional and post-translational changes that increase titin’s length and extensibility, making the sarcomere longer and softer, are associated with systolic dysfunction and left ventricular dilation. Modifications of titin that decrease its length and extensibility, making the sarcomere shorter and stiffer, are associated with diastolic dysfunction in animal models. There has been significant progress in understanding the mechanisms by which titin is modified. As molecular pathways that modify titin’s mechanical properties are elucidated, they represent therapeutic targets for treatment of both systolic and diastolic dysfunction. In this article, we review titin’s contribution to normal cardiac physiology, the pathophysiology of titin truncation variations leading to dilated cardiomyopathy, and transcriptional and post-translational modifications of titin. Emphasis is on how modification of titin can be utilized as a therapeutic target for treatment of heart failure.

Abstract MP105: Python Post-prandial Cardiac Adaptation is Supported by Enhanced Metabolism, Reduced Sarcomere Passive Tension, and Epigenetic Remodeling

Pythons are infrequent feeders that can ingest meals equal to their own body mass. The extreme metabolic response required to digest such large meals is associated with a dramatic increase in the mass of most organs, including the heart. Recently, we have been able to assess functional effects of feeding using isolated python cardiomyocytes and myofibrils, advancing our understanding of extreme cardiac adaptation in python ( Python regius ). Twenty-four hours after feeding, python cardiomyocytes showed prolonged Ca 2+ transients, increased maximal tension and Ca 2+ sensitivity of myofibrils as compared to fasted pythons. Post-prandial positive inotropy was accompanied by enhanced metabolic output via increased mitochondrial ATP production rate and by AMP-dependent kinase (AMPK) activation and phosphofructokinase-2 reduction, suggesting a key role for fatty acid, but not glucose, metabolism after feeding. In addition, 24h post-fed hearts had significantly reduced tissue stiffness and myofibril passive tension. Finally, chromatin condensation was reduced about 30% after feeding in python cardiomyocytes and confirmed by increased histone acetylation, indicating a predominant role for epigenetics in post-prandial adaptation. These results suggest that feeding promotes positive cardiac inotropy in python via a number of coordinated mechanisms to enhance energy production, increase myofibril and tissue compliance, and increase chromatin accessibility. As heart failure is commonly characterized by depressed contractility, compromised energetics, and increased tissue stiffness, assessing post-prandial adaptation in python hearts provides us with powerful insights that could inform the development of therapeutics for human heart diseases.

Pathophysiology of exercise-induced muscle damage and its structural, functional, metabolic, and clinical consequences

Extreme or unaccustomed eccentric exercise can cause exercise-induced muscle damage, characterized by structural changes involving sarcomere, cytoskeletal, and membrane damage, with an increased permeability of sarcolemma for proteins. From a functional point of view, disrupted force transmission, altered calcium homeostasis, disruption of excitation-contraction coupling, as well as metabolic changes bring about loss of strength. Importantly, the trauma also invokes an inflammatory response and clinically presents itself by swelling, decreased range of motion, increased passive tension, soreness, and a transient decrease in insulin sensitivity. While being damaging and influencing heavily the ability to perform repeated bouts of exercise, changes produced by exercise-induced muscle damage seem to play a crucial role in myofibrillar adaptation. Additionally, eccentric exercise yields greater hypertrophy than isometric or concentric contractions and requires less in terms of metabolic energy and cardiovascular stress, making it especially suitable for the elderly and people with chronic diseases. This review focuses on our current knowledge of the mechanisms underlying exercise-induced muscle damage, their dependence on genetic background, as well as their consequences at the structural, functional, metabolic, and clinical level. A comprehensive understanding of these is a prerequisite for proper inclusion of eccentric training in health promotion, rehabilitation, and performance enhancement.

Novel compact device for clinically measuring extraocular muscle (EOM) tension

Accurate knowledge of extraocular muscle (EOM) tension is important for the diagnosis of and surgical planning for strabismus, such as choosing which eye to operate or determining the amount of muscle displacement. Previous evaluations of passive EOM tension have relied extensively on the experience and skill of ophthalmic surgeons, who generally perform such evaluations by gripping the eyeball and then pushing and pulling it. This methodology, named the forced duction test, has the significant limitation that the tension is felt subjectively via the forceps, with the results therefore not being quantifiable. Previous quantitative analyses have utilized several different types of equipment with implanted force transducers or have involved connecting the muscle tendon to a strain gauge. However, the associated equipment setups and recording systems are highly complex and rarely used outside research settings. This situation prompted the present study to develop a novel compact, quantifiable and clinically applicable device for measuring the passive tension in human EOMs for use in clinical practice. The device employs locking forceps and a tilting sensor to rule out effects of the gripping force and to compensate for changes in the force due to tilting, which improves the measurement accuracy. The performance of the device was investigated in 60 eyes of 30 consecutive anaesthetized patients immediately prior to ophthalmic surgery. The results showed that the measured EOM tension in each rectus muscles agreed with previous findings: 48.3 ± 14.5 g (0.82 ± 0.28 g/deg, mean ± SD) for the lateral rectus, 45.6 ± 13.2 g (0.82 ± 0.23 g/deg) for the medial rectus, 48.6 ± 14.7 g (0.71 ± 0.21 g/deg) for the inferior rectus and 53.4 ± 13.7 g (0.77 ± 0.25 g/deg) for the superior rectus.

Alterations of Extracellular Matrix Mechanical Properties Contribute to Age-Related Functional Impairment of Human Skeletal Muscles.

Aging of human skeletal muscles is associated with increased passive stiffness, but it is still debated whether muscle fibers or extracellular matrix (ECM) are the determinants of such change. To answer this question, we compared the passive stress generated by elongation of fibers alone and arranged in small bundles in young healthy (Y: 21 years) and elderly (E: 67 years) subjects. The physiological range of sarcomere length (SL) 2.5–3.3 μm was explored. The area of ECM between muscle fibers was determined on transversal sections with picrosirius red, a staining specific for collagen fibers. The passive tension of fiber bundles was significantly higher in E compared to Y at all SL. However, the resistance to elongation of fibers alone was not different between the two groups, while the ECM contribution was significantly increased in E compared to Y. The proportion of muscle area occupied by ECM increased from 3.3% in Y to 8.2% in E. When the contribution of ECM to bundle tension was normalized to the fraction of area occupied by ECM, the difference disappeared. We conclude that, in human skeletal muscles, the age-related reduced compliance is due to an increased stiffness of ECM, mainly caused by collagen accumulation.

Deleting Titin's C-Terminal PEVK Exons Increases Passive Stiffness, Alters Splicing, and Induces Cross-Sectional and Longitudinal Hypertrophy in Skeletal Muscle.

The Proline, Glutamate, Valine and Lysine-rich (PEVK) region of titin constitutes an entropic spring that provides passive tension to striated muscle. To study the functional and structural repercussions of a small reduction in the size of the PEVK region, we investigated skeletal muscles of a mouse with the constitutively expressed C-terminal PEVK exons 219–225 deleted, the TtnΔ219–225 model (MGI: TtnTM 2.1Mgot). Based on this deletion, passive tension in skeletal muscle was predicted to be increased by ∼17% (sarcomere length 3.0 μm). In contrast, measured passive tension (sarcomere length 3.0 μm) in both soleus and EDL muscles was increased 53 ± 11% and 62 ± 4%, respectively. This unexpected increase was due to changes in titin, not to alterations in the extracellular matrix, and is likely caused by co-expression of two titin isoforms in TtnΔ219–225 muscles: a larger isoform that represents the TtnΔ219–225 N2A titin and a smaller isoform, referred to as N2A2. N2A2 represents a splicing adaption with reduced expression of spring element exons, as determined by titin exon microarray analysis. Maximal tetanic tension was increased in TtnΔ219–225 soleus muscle (WT 240 ± 9; TtnΔ219–225 276 ± 17 mN/mm2), but was reduced in EDL muscle (WT 315 ± 9; TtnΔ219–225 280 ± 14 mN/mm2). The changes in active tension coincided with a switch toward slow fiber types and, unexpectedly, faster kinetics of tension generation and relaxation. Functional overload (FO; ablation) and hindlimb suspension (HS; unloading) experiments were also conducted. TtnΔ219–225 mice showed increases in both longitudinal hypertrophy (increased number of sarcomeres in series) and cross-sectional hypertrophy (increased number of sarcomeres in parallel) in response to FO and attenuated cross-sectional atrophy in response to HS. In summary, slow- and fast-twitch muscles in a mouse model devoid of titin’s PEVK exons 219–225 have high passive tension, due in part to alterations elsewhere in splicing of titin’s spring region, increased kinetics of tension generation and relaxation, and altered trophic responses to both functional overload and unloading. This implicates titin’s C-terminal PEVK region in regulating passive and active muscle mechanics and muscle plasticity.

Effect of Parenting Training on Neurobehavioral Development of Infants

BackgroundEarly interventions have been believed to have a positive influence on the neurodevelopment of infants. Our Child Health Center has carried out parenting training for parents of infants for several years to promote the neurobehavioral development of infants at an early stage.Material/MethodsWe enrolled 117 families with term infants age 0–3 months who had completed a parenting training class at the Child Health Center of the Department of Pediatrics, the Third Xiangya Hospital. Parenting training included 4 parts: nursing, intelligence, social contact, and physical ability. A nurse practitioner demonstrated procedures to parents, who then performed them at home for 1 month. The Neonatal Behavioral Neurological Assessment (NBNA) was used to evaluate infants before and 1 month after parenting training.ResultsIn the comparative analysis before and after parenting training, there was a significant increase in the NBNA scores. For the infants whose parents received parenting training, the NBNA scores in total score (33.74±0.19 before parenting training vs. 36.69±0.20 after 1 month), neonatal behavioral capacity (10.19±0.14 before parenting training vs. 11.26±0.10 after 1 month), passive muscle tension (7.28±0.07 before parenting training vs. 7.82±0.04 after 1 month), and initiative muscle tension (4.29±0.08 before the parenting training vs. 5.61±0.13 after 1 month) were significantly higher one month before (P<0.01).ConclusionsTerm infant neurobehavior was associated with participation in parenting training, suggesting that these practices of parenting training support better early neurobehavioral development of infants.

Effects of fasting on isolated murine skeletal muscle contractile function during acute hypoxia.

Stored muscle carbohydrate supply and energetic efficiency constrain muscle functional capacity during exercise and are influenced by common physiological variables (e.g. age, diet, and physical activity level). Whether these constraints affect overall functional capacity or the timing of muscle energetic failure during acute hypoxia is not known. We interrogated skeletal muscle contractile properties in two anatomically distinct rodent hindlimb muscles that have well characterized differences in energetic efficiency (locomotory- extensor digitorum longus (EDL) and postural- soleus muscles) following a 24 hour fasting period that resulted in substantially reduced muscle carbohydrate supply. 180 mins of acute hypoxia resulted in complete energetic failure in all muscles tested, indicated by: loss of force production, substantial reductions in total adenosine nucleotide pool intermediates, and increased adenosine nucleotide degradation product-inosine monophosphate (IMP). These changes occurred in the absence of apparent myofiber structural damage assessed histologically by both transverse section and whole mount. Fasting and the associated reduction of the available intracellular carbohydrate pool (~50% decrease in skeletal muscle) did not significantly alter the timing to muscle functional impairment or affect the overall force/work capacities of either muscle type. Fasting resulted in greater passive tension development in both muscle types, which may have implications for the design of pre-clinical studies involving optimal timing of reperfusion or administration of precision therapeutics.

Carotid Artery Reactivity in Young, Adult, and Aged APOE3 and APOE4 Mice

IntroductionAlzheimer’s disease (AD) is the most common form of dementia and is the 6th leading cause of death in the US. Growing evidence supports a link between AD and cardiovascular disease (CVD). Interestingly, variant isoforms of apolipoprotein E (APOE) are risk factors for both AD and CVD. The most common APOE isoform, APOE3, has a neutral risk for developing AD, whereas APOE4 increases AD risk. In order to study APOE‐mediated processes in large arteries, carotid arteries (CAs) were isolated from mice expressing human‐ ApoE targeted replacement of APOE3 (B6.129P2‐Apoetm2(APOE*3)Mae N8) and APOE4 (B6.129P2‐Apoetm3(APOE*4)Mae N8)(Taconic Labs). Our objective was to determine whether allelic expression alters CA vascular function by assessing mechanical characteristics and vascular reactivity in the left main CA isolated from young (Y, 3mo), adult (A, 12mo) and aged (Aged, 18–22mo), homozygous APOE3 and APOE4, male and female mice. We hypothesize that APOE4 expression alters CA vascular reactivity, along with vascular structure compared to CAs isolated from age matched APOE3 mice.MethodsCAs were rapidly isolated from age matched male and female, APOE3 and APOE4 mice, cleaned of connective tissue, cut into 8mm segments, and cannulated in an arteriograph chamber to assess vascular reactivity via myogenic tone (MT) and phenylephrine (PE)‐induced constriction in the absence and presence of the non‐specific NOS inhibitor, L‐NAME. In addition, passive wall tension, arterial wall thickness, vascular distensibility, and arterial stress/strain were measured at intraluminal pressures ranging from 10mm Hg to 140mm Hg for assessment of age‐related changes in arterial structure/function.ResultsCAs from APOE3 and APOE4 mice did not exhibit MT in the absence or presence of L‐NAME. PE‐induced constriction in CA from APOE3 mice tended to decrease with age, while PE‐induced constriction in CA from A APOE4 mice were significantly greater than PE‐induced constriction in CAs from Y (P&lt;0.05) or Aged (P&lt;0.05) mice. In the presence of L‐NAME, PE‐induced constrictions were significantly increased in all mice at all ages (P&lt;0.05). Wall thickness was similar in Y, A and Aged APOE3 and APOE 4 mice. Finally, CA from Y APOE3 mice exhibited a rightward shift in stress vs strain, compared to CAs from A and Aged mice. In contrast, CAs from Y and A, APOE4 mice exhibited similar stress vs strain values. Stress vs strain values in Aged APOE4 CAs were significantly shifted to the left of CA from A APOE4 mice suggesting greater arterial stiffening in Aged APOE4 mice at physiologically relevant intraluminal pressures (P&lt;0.05).Summary/ConclusionsThese data suggest that CA vascular structure/function in APOE3 mice exhibit “normal” aging characteristics, whereas CAs from APOE4 mice exhibit “delayed” age related changes in arterial stiffening, which may contribute to elevated cerebral blood flow in Y human and mice carrying the APOE4 allele vs aged matched APOE3.Support or Funding InformationFundingABRC/ADHS18‐205211 (DME, JVE, CJ, TBJ), Biomedical Sciences Program (LJS, RGP, DME), Biomedical Sciences Start‐up Funds (DME)

The relationships between passive tension of quadriceps muscle and knee flexion in female patients with knee osteoarthritis

Purpose: Reduction in knee flexion excursion has been reported during the early stance phase of gait in patients with knee osteoarthritis (KOA). Quadriceps femoris muscle is the major muscle group that functions eccentrically in controlling the knee flexion motion during the early loading phase. Since force generated by muscle is influenced by both motor unit activation and muscle stiffness, we were interested to explore possible associations between knee flexion motion and passive property of the quadriceps femoris muscle.

Non-linear Scaling of Passive Mechanical Properties in Fibers, Bundles, Fascicles and Whole Rabbit Muscles.

Defining variations in skeletal muscle passive mechanical properties at different size scales ranging from single muscle fibers to whole muscles is required in order to understand passive muscle function. It is also of interest from a muscle structural point-of-view to identify the source(s) of passive tension that function at each scale. Thus, we measured passive mechanical properties of single fibers, fiber bundles, fascicles, and whole muscles in three architecturally diverse muscles from New Zealand White rabbits (n = 6) subjected to linear deformation. Passive modulus was quantified at sarcomere lengths across the muscle’s anatomical range. Titin molecular mass and collagen content were also quantified at each size scale, and whole muscle architectural properties were measured. Passive modulus increased non-linearly from fiber to whole muscle for all three muscles emphasizing extracellular sources of passive tension (p < 0.001), and was different among muscles (p < 0.001), with significant muscle by size-scale interaction, indicating quantitatively different scaling for each muscle (p < 0.001). These findings provide insight into the structural basis of passive tension and suggest that the extracellular matrix (ECM) is the dominant contributor to whole muscle and fascicle passive tension. They also demonstrate that caution should be used when inferring whole muscle properties from reduced muscle size preparations such as muscle biopsies.

Intact myocardial preparations reveal intrinsic transmural heterogeneity in cardiac mechanics

Determining transmural mechanical properties in the heart provides a foundation to understand physiological and pathophysiological cardiac mechanics. Although work on mechanical characterisation has begun in isolated cells and permeabilised samples, the mechanical profile of living individual cardiac layers has not been examined. Myocardial slices are 300 μm-thin sections of heart tissue with preserved cellular stoichiometry, extracellular matrix, and structural architecture. This allows for cardiac mechanics assays in the context of an intact in vitro organotypic preparation. In slices obtained from the subendocardium, midmyocardium and subepicardium of rats, a distinct pattern in transmural contractility is found that is different from that observed in other models. Slices from the epicardium and midmyocardium had a higher active tension and passive tension than the endocardium upon stretch. Differences in total myocyte area coverage, and aspect ratio between layers underlined the functional readouts, while no differences were found in total sarcomeric protein and phosphoprotein between layers. Such intrinsic heterogeneity may orchestrate the normal pumping of the heart in the presence of transmural strain and sarcomere length gradients in the in vivo heart.

Determining the Optimal Normalization Factor of Different Target Arteries for ex vivo Vascular Function Experiments: A New Standardized Procedure

The standardization of resistance vessel preparation is crucial to compare physiologic vascular reactivity under different experimental conditions. Here, we describe a generalizable experimental setup for ex vivo vascular function experiments and their mathematical basis. Porcine basilar arteries and chicken common carotid arteries were isolated post mortem via standardized surgical approaches. The inner circumference of these vessels with a passive wall tension corresponding to 100 mm Hg (IC₁₀₀) as well as the circumference at which the active force production of the vessel is maximal (IC₁) were determined systematically. The IC₁/IC₁₀₀ ratio (also referred to as factor k), a value that is believed to be constant for a defined vessel type in one species, was calculated by a novel mathematical approach. Here, we present an easy-to-use toolbox for the systematic and computer-based calculation of factor k and simplified optimal pre-stretching of any vascular segments for wire myography experiments.