Michael A. Petrie

A minimal dose of electrically induced muscle activity regulates distinct gene signaling pathways in humans with spinal cord injury.

Paralysis after a spinal cord injury (SCI) induces physiological adaptations that compromise the musculoskeletal and metabolic systems. Unlike non-SCI individuals, people with spinal cord injury experience minimal muscle activity which compromises optimal glucose utilization and metabolic control. Acute or chronic muscle activity, induced through electrical stimulation, may regulate key genes that enhance oxidative metabolism in paralyzed muscle. We investigated the short and long term effects of electrically induced exercise on mRNA expression of human paralyzed muscle. We developed an exercise dose that activated the muscle for only 0.6% of the day. The short term effects were assessed 3 hours after a single dose of exercise, while the long term effects were assessed after training 5 days per week for at least one year (adherence 81%). We found a single dose of exercise regulated 117 biological pathways as compared to 35 pathways after one year of training. A single dose of electrical stimulation increased the mRNA expression of transcriptional, translational, and enzyme regulators of metabolism important to shift muscle toward an oxidative phenotype (PGC-1α, NR4A3, IFRD1, ABRA, PDK4). However, chronic training increased the mRNA expression of specific metabolic pathway genes (BRP44, BRP44L, SDHB, ACADVL), mitochondrial fission and fusion genes (MFF, MFN1, MFN2), and slow muscle fiber genes (MYH6, MYH7, MYL3, MYL2). These findings support that a dose of electrical stimulation (∼10 minutes/day) regulates metabolic gene signaling pathways in human paralyzed muscle. Regulating these pathways early after SCI may contribute to reducing diabetes in people with longstanding paralysis from SCI.

Low-frequency stimulation regulates metabolic gene expression in paralyzed muscle

The altered metabolic state after a spinal cord injury compromises systemic glucose regulation. Skeletal muscle atrophies and transforms into fast, glycolytic, and insulin-resistant tissue. Osteoporosis is common after spinal cord injury and limits the ability to exercise paralyzed muscle. We used a novel approach to study the acute effect of two frequencies of stimulation (20 and 5 Hz) on muscle fatigue and gene regulation in people with chronic paralysis. Twelve subjects with chronic (>1 yr) and motor complete spinal cord injury (ASIA A) participated in the study. We assessed the twitch force before and after a single session of electrical stimulation (5 or 20 Hz). We controlled the total number of pulses delivered for each protocol (10,000 pulses). Three hours after the completion of the electrical stimulation (5 or 20 Hz), we sampled the vastus lateralis muscle and examined genes involved with metabolic transcription, glycolysis, oxidative phosphorylation, and mitochondria remodeling. We discovered that the 5-Hz stimulation session induced a similar amount of fatigue and a five- to sixfold increase (P < 0.05) in key metabolic transcription factors, including PGC-1α, NR4A3, and ABRA as the 20-Hz session. Neither session showed a robust regulation of genes for glycolysis, oxidative phosphorylation, or mitochondria remodeling. We conclude that a low-force and low-frequency stimulation session is effective at inducing fatigue and regulating key metabolic transcription factors in human paralyzed muscle. This strategy may be an acceptable intervention to improve systemic metabolism in people with chronic paralysis.

Low force contractions induce fatigue consistent with muscle mRNA expression in people with spinal cord injury

Spinal cord injury (SCI) is associated with muscle atrophy, transformation of muscle fibers to a fast fatigable phenotype, metabolic inflexibility (diabetes), and neurogenic osteoporosis. Electrical stimulation of paralyzed muscle may mitigate muscle metabolic abnormalities after SCI, but there is a risk for a fracture to the osteoporotic skeletal system. The goal of this study was to determine if low force stimulation (3 Hz) causes fatigue of chronically paralyzed muscle consistent with selected muscle gene expression profiles. We tested 29 subjects, nine with a SCI and 20 without and SCI, during low force fatigue protocol. Three SCI and three non‐SCI subjects were muscle biopsied for gene and protein expression analysis. The fatigue index (FI) was 0.21 ± 0.27 and 0.91 ± 0.01 for the SCI and non‐SCI groups, respectively, supporting that the low force protocol physiologically fatigued the chronically paralyzed muscle. The post fatigue potentiation index (PI) for the SCI group was increased to 1.60 ± 0.06 (P < 0.001), while the non‐SCI group was 1.26 ± 0.02 supporting that calcium handling was compromised with the low force stimulation. The mRNA expression from genes that regulate atrophy and fast properties (MSTN, ANKRD1, MYH8, and MYCBP2) was up regulated, while genes that regulate oxidative and slow muscle properties (MYL3, SDHB, PDK2, and RyR1) were repressed in the chronic SCI muscle. MSTN, ANKRD1, MYH8, MYCBP2 gene expression was also repressed 3 h after the low force stimulation protocol. Taken together, these findings support that a low force single twitch activation protocol induces paralyzed muscle fatigue and subsequent gene regulation. These findings suggest that training with a low force protocol may elicit skeletal muscle adaptations in people with SCI.

Distinct skeletal muscle gene regulation from active contraction, passive vibration, and whole body heat stress in humans

Skeletal muscle exercise regulates several important metabolic genes in humans. We know little about the effects of environmental stress (heat) and mechanical stress (vibration) on skeletal muscle. Passive mechanical stress or systemic heat stress are often used in combination with many active exercise programs. We designed a method to deliver a vibration stress and systemic heat stress to compare the effects with active skeletal muscle contraction. Purpose: The purpose of this study is to examine whether active mechanical stress (muscle contraction), passive mechanical stress (vibration), or systemic whole body heat stress regulates key gene signatures associated with muscle metabolism, hypertrophy/atrophy, and inflammation/repair. Methods: Eleven subjects, six able-bodied and five with chronic spinal cord injury (SCI) participated in the study. The six able-bodied subjects sat in a heat stress chamber for 30 minutes. Five subjects with SCI received a single dose of limb-segment vibration or a dose of repetitive electrically induced muscle contractions. Three hours after the completion of each stress, we performed a muscle biopsy (vastus lateralis or soleus) to analyze mRNA gene expression. Results: We discovered repetitive active muscle contractions up regulated metabolic transcription factors NR4A3 (12.45 fold), PGC-1α (5.46 fold), and ABRA (5.98 fold); and repressed MSTN (0.56 fold). Heat stress repressed PGC-1α (0.74 fold change; p < 0.05); while vibration induced FOXK2 (2.36 fold change; p < 0.05). Vibration similarly caused a down regulation of MSTN (0.74 fold change; p < 0.05), but to a lesser extent than active muscle contraction. Vibration induced FOXK2 (p < 0.05) while heat stress repressed PGC-1α (0.74 fold) and ANKRD1 genes (0.51 fold; p < 0.05). Conclusion: These findings support a distinct gene regulation in response to heat stress, vibration, and muscle contractions. Understanding these responses may assist in developing regenerative rehabilitation interventions to improve muscle cell development, growth, and repair.

Effects of brain derived neurotrophic factor Val66Met polymorphism in patients with cervical spondylotic myelopathy.

Cervical spondylotic myelopathy (CSM) is the leading cause of spinal cord related disability in the elderly. It results from degenerative narrowing of the spinal canal, which causes spinal cord compression. This leads to gait instability, loss of dexterity, weakness, numbness and urinary dysfunction. There has been indirect data that implicates a genetic component to CSM. Such a finding may contribute to the variety in presentation and outcome in this patient population. The Val66Met polymorphism, a mutation in the brain derived neurotrophic factor (BDNF) gene, has been implicated in a number of brain and psychological conditions, and here we investigate its role in CSM. Ten subjects diagnosed with CSM were enrolled in this prospective study. Baseline clinical evaluation using the modified Japanese Orthopaedic Association (mJOA) scale, Nurick and 36-Item Short Form Health Survey (SF-36) were collected. Each subject underwent objective testing with gait kinematics, as well as hand functioning using the Purdue Peg Board. Blood samples were analyzed for the BDNF Val66Met mutation. The prevalence of the Val66Met mutation in this study was 60% amongst CSM patients compared to 32% in the general population. Individuals with abnormal Met allele had worse baseline mJOA and Nurick scores. Moreover, baseline gait kinematics and hand functioning testing were worse compared to their wild type counterpart. BDNF Val66Met mutation has a higher prevalence in CSM compared to the general population. Those with BDNF mutation have a worse clinical presentation compared to the wild type counterpart. These findings suggest implication of the BDNF mutation in the development and severity of CSM.

Vibration training after chronic spinal cord injury: Evidence for persistent segmental plasticity

H-reflex paired-pulse depression is gradually lost within the first year post-SCI, a process believed to reflect reorganization of segmental interneurons after the loss of normal descending (cortical) inhibition. This reorganization co-varies in time with the development of involuntary spasms and spasticity. The purpose of this study is to determine whether long-term vibration training may initiate the return of H-reflex paired-pulse depression in individuals with chronic, complete SCI. Five men with SCI received twice-weekly vibration training (30Hz, 0.6g) to one lower limb while seated in a wheelchair. The contra-lateral limb served as a within-subject control. Paired-pulse H-reflexes were obtained before, during, and after a session of vibration. Untrained limb H-reflex depression values were comparable to chronic SCI values from previous reports. In contrast, the trained limbs of all 5 participants showed depression values that were within the range of previously-reported Acute SCI and Non-SCI H-reflex depression. The average difference between limbs was 34.98% (p=0.016). This evidence for the return of H-reflex depression suggests that even for people with long-standing SCI, plasticity persists in segmental reflex pathways. The spinal networks involved with the clinical manifestation of spasticity may thus retain adaptive plasticity after long-term SCI. The results of this study indicate that vibration training may hold promise as an anti-spasticity rehabilitation intervention.

Effect of Surgery on Gait and Sensory Motor Performance in Patients With Cervical Spondylotic Myelopathy.

BACKGROUND Cervical spondylotic myelopathy (CSM) is a common disease of aging that leads to gait instability resulting from loss of leg sensory and motor functions. The results of surgical intervention have been studied using a variety of methods, but no test has been reported that objectively measures integrative leg motor sensory functions in CSM patients. OBJECTIVE To determine the feasibility of using a novel single leg squat (SLS) test to measure integrative motor sensory functions in patients with CSM before and after surgery. METHODS Fifteen patients with CSM were enrolled in this prospective study. Clinical data and scores from standard outcomes questionnaires were obtained before and after surgery. Patients also participated in experimental test protocols consisting of standard kinematic gait testing, the Purdue pegboard test, and the novel SLS test. RESULTS The SLS test protocol was well tolerated by CSM patients and generated objective performance data over short test periods. In patients who participated in postoperative testing, the group measures of mean SLS errors decreased following surgery. Gait velocity measures followed a similar pattern of group improvement postoperatively. Practical barriers to implementing this extensive battery of tests resulted in subject attrition over time. Compared with kinematic gait testing, the SLS protocol required less space and could be effectively implemented more efficiently. CONCLUSIONS The SLS test provides a practical means of obtaining objective measures of leg motor sensory functions in patients with CSM. Additional testing with a larger cohort of patients is required to use SLS data to rigorously examine group treatment effects. ABBREVIATIONS BW, body weightCSM, cervical spondylotic myelopathymJOA, modified Japanese Orthopedic AssociationSLS, single leg squat.

Mechanical properties of the haptic signals indicative of a breast cancer tumor

A clinical breast exam (CBE), in which a nurse or doctor palpates a patients breast tissue searching for hard lumps, is a recommended annual breast cancer diagnostic for women over 40. Recent advances in training technology have suggested that the sensitivity of this exam can be improved and that clinicians are interested in improving training approaches. Currently researchers have an incomplete understanding of which features of the force pattern an observer must perceive in order to recognize the presence of a tumor. This limitation in our knowledge limits our ability to build more effective simulators that emphasize the specific characteristics of the signal that clinicians must perceive. Three experiments were performed using tissue analogs made from silicone rubber embedded with hard spheres. The size of the balls, their depth and the stiffness of the silicone were varied. The force exerted on a probe indented at regular intervals along the samples revealed the general force profile that a clinician may experience on his or her finger pad when searching for a tumor. Several measures of the force profiles were compared to the salience of the ball within the silicone: the ratio of the response force directly above the sphere to the force response far from the sphere, the rate of change of the force response near the sphere, and difference between the force above and to the side divided by the force to the side. These measures correlated with salience when the ball size and ball depth was varied, but not when the silicone stiffness varied. The results suggest that the relationship between the force profile and the salience of the stimulus is more complex that expected. Once this relationship is more fully understood, new training tools and procedures can be developed to trains clinicians and improve the sensitivity of clinical breast exams.

Whole body heat exposure modulates acute glucose metabolism

Abstract Aim: Exercise modulates glucose tolerance and homeostasis in both healthy and diabetic individuals. Heat stress is a fundamental element of exercise. The acute glycemic response and alterations in glucose clearance following whole body passive heat stress in the absence of muscle activity has yet to be examined in humans. Knowledge of this relationship may prove useful, particularly in populations with compromised glucoregulation from reduced activity. Purpose: To determine insulin/glucose levels before and after an acute bout of heat stress in healthy, lean individuals and examine the effects of whole body heat stress (WBHS) and exercise on acute glucose tolerance in an expanded cohort. Methods: Ten subjects (24.1 ± 0.7 years) participated in a randomized control/WBHS session (up to 30 minutes at 73 °C) with fasting glucose (FG) and insulin drawn at baseline, immediately after and 30 minutes post heat stress. In the follow-up experiment, 20 anthropometrically diverse subjects (24.6 ± 2.1 years) underwent an oral glucose tolerance test (OGTT) under the conditions above. Results: FG levels rose 10% immediately following heat stress (8.6 (±5.6) mg/dl, p < .01) and returned to near baseline levels 30 minutes following WBHS. Insulin release showed its greatest increase at 30 minutes post WBHS (2.7 ± 3.5) uU/ml p < .05). WBHS resulted in a decrease in glucose uptake [AUC increased 8.2% (1430.6 ± 1957.03) mg/dl (p = .005)], particularly in nonlean individuals. Conclusion: WBHS modulates physiologic markers of metabolism. An acute bout of WBHS increases glucose and insulin levels in healthy individual and decreases glucose uptake in response to a glucose challenge, particularly those who are non-lean.

Speed, resistance, and unexpected accelerations modulate feed forward and feedback control during a novel weight bearing task

We developed a method to investigate feed-forward and feedback movement control during a weight bearing visuomotor knee tracking task. We hypothesized that a systematic increase in speed and resistance would show a linear decrease in movement accuracy, while unexpected perturbations would induce a velocity-dependent decrease in movement accuracy. We determined the effects of manipulating the speed, resistance, and unexpected events on error during a functional weight bearing task. Our long term objective is to benchmark neuromuscular control performance across various groups based on age, injury, disease, rehabilitation status, and/or training. Twenty-six healthy adults between the ages of 19-45 participated in this study. The study involved a single session using a custom designed apparatus to perform a single limb weight bearing task under nine testing conditions: three movement speeds (0.2, 0.4, and 0.6Hz) in combination with three levels of brake resistance (5%, 10%, and 15% of individuals body weight). Individuals were to perform the task according to a target with a fixed trajectory across all speeds, corresponding to a∼0 (extension) to 30° (flexion) of knee motion. An increase in error occurred with speed (p<0.0001, effect size (eta2): η2=0.50) and resistance (p<0.0001, η2=0.01). Likewise, during unexpected perturbations, the ratio of perturbed/non-perturbed error increased with each increment in velocity (p<0.0014, η2=0.08), and resistance (p<0.0001, η2=0.11). The hierarchical framework of these measurements offers a standardized functional weight bearing strategy to assess impaired neuro-muscular control and/or test the efficacy of therapeutic rehabilitation interventions designed to influence neuromuscular control of the knee.