Publications

Neurogenic bladder (NB) affects people of all ages. Electric impedance myography (EIM) assesses localized muscle abnormalities. Here, we sought to investigate whether unique detrusor EIM signatures are present in NB due to spinal cord injury (SCI). Twenty-eight, 8-10 weeks old, C57BL/6J female mice were studied. Twenty underwent spinal cord transection; 8 served as controls. Cohorts were euthanized at 4 and 6 weeks after spinal cord transection. Each bladder was measured in-situ with EIM with applied frequencies of 1 kHz to 10 MHz, and then processed for molecular and histologic study. SCI mice had greater bladder-to-body weight ratio (p < 0.0001), greater collagen deposition (p = 0.009), and greater smooth-muscle-myosin-heavy-chain isoform A/B ratio (p < 0.0001). Compared with the control group, the SCI group was associated with lower phase, reactance, and resistance values (p < 0.01). Significant correlations (p < 0.001) between bladder-to-body weight ratios and EIM measurements were observed across the entire frequency spectrum. A severely hypertrophied phenotype was characterized by even greater bladder-to-body weight ratios and more depressed EIM values. Our study demonstrated distinct EIM alterations in the detrusor muscle of mice with NB due to SCI. With further refinement, EIM may offer a potential point-of-care tool for the assessment of NB and its response to treatment.

Skeletal muscle size and strength are important for overall health for astronauts. However, how male and female muscle may respond differently to micro- and partial-gravity environments is not fully understood. The purpose of this study was to determine how biological sex and sex steroid hormones influence the progression of muscle atrophy after long term exposure to micro and partial gravity environments in male and female rats. Male and female Fisher rats (n ​= ​120) underwent either castration/ovariectomy or sham surgeries. After two weeks recovery, animals were divided into microgravity (0g), partial-gravity (40% of weight bearing, 0.4g), or full weight bearing (1g) interventions for 28 days. Measurements of muscle size and strength were evaluated prior to and after interventions. At 0g, females lost more dorsiflexion strength, plantar flexion strength, and other metrics of muscle size compared to males; castration/ovariectomy did not influence these differences. Additionally, at 0.4g, females lost more dorsiflexion strength, plantar flexion strength, and other metrics of muscle strength compared to males; castration/ovariectomy did not influence these differences. Females have greater musculoskeletal aberrations during exposure to both microgravity and partial-gravity environments; these differences are not dependent on the presence of sex steroid hormones. Correspondingly, additional interventions may be necessary to mitigate musculoskeletal loss in female astronauts to protect occupational and overall health.

The Harvard Catalyst KL2/CMeRIT program is a 2-year mentored institutional career award that includes KL2 grants funded by National Institutes of Health (NIH) and CMeRIT grants funded by Harvard Catalyst nonfederal funds. The purpose of this study was to compare outcomes for early-stage investigators funded by the KL2/CMeRIT program to a group of applicants who were not chosen for support to assess the potential impact of the program on early career outcomes. Career data, including academic promotions, subsequent grant funding, and publication rates, from both successful and unsuccessful 2008-2018 KL2/CMeRIT applicants were compiled throughout the year 2020. Data were obtained directly through outreach to both groups and through assessment of online resources. The cohort comprised 487 individuals, 109 awardees, and 378 nonawardees. Awardees were more likely to be subsequently involved in clinical and translational research than nonawardees (92% vs 75%, p < 0.001). A higher proportion of awardees also had achieved academic promotion (81% vs 69%, p = 0.016) and subsequent NIH funding (72% vs 58%, p = 0.047), while there was no difference in publication rates (p = 0.555). Participants in the Harvard Catalyst KL2/CMeRIT program demonstrate greater early career success than nonparticipants though the nonparticipants also fared relatively well.

INTRODUCTION/AIMS: Needle impedance-electromyography (iEMG) is a diagnostic modality currently under development that combines intramuscular electrical impedance with concentric electromyography (EMG) in a single needle. We designed, manufactured, and tested a prototype iEMG needle in a cohort of wild-type (WT) and SOD1G93A amyotrophic lateral sclerosis (ALS) mice to assess its ability to record impedance and EMG data.

METHODS: A new six-electrode, 26-gauge, iEMG needle was designed, manufactured and tested. Quantitative impedance and qualitative "gestalt" EMG were performed sequentially on bilateral quadriceps of 16-wk-old SOD1G93A ALS (N = 6) and WT (N = 6) mice by connecting the needle first to an impedance analyzer (with the animal at rest) and then to a standard EMG system (with the animal fully under anesthesia to measure spontaneous activity and briefly during awakening to measure voluntary activity). The needle remained in the muscle throughout the measurement period.

RESULTS: EMG data were qualitatively similar to that observed with a commercially available concentric EMG needle; fibrillation potentials were observed in 84% of the ALS mice and none of the WT mice; motor unit potentials were also readily identified. Impedance data revealed significant differences in resistance, reactance, and phase values between the two groups, with ALS animals having reduced reactance and resistance values.

DISCUSSION: This work demonstrates the feasibility of a single iEMG needle conforming to standard dimensions of size and function. Further progress of iEMG technology for enhanced neuromuscular diagnosis and quantification of disease status is currently in development.

Electrical impedance methods, including electrical impedance myography, are increasingly being used as biomarkers of muscle health since they measure passive electrical properties of muscle that alter in disease. One disorder, Pompe Disease (Glycogen storage disease type II (GSDII)), remains relatively unstudied. This disease is marked by dramatic accumulation of intracellular myofiber glycogen. Here we assessed the electrical properties of skeletal muscle in a model of GSDII, the Pompe6neo/6neo (Pompe) mouse. Ex vivo impedance measurements of gastrocnemius (GA) were obtained using a dielectric measuring cell in 30-week-old female Pompe (N = 10) and WT (N = 10) mice. Longitudinal and transverse conductivity, σ, and the relative permittivity, εr, and Cole-Cole complex resistivity parameters at 0 Hz and infinite frequency, ρo and ρ∞, respectively, and the intracellular resistivity, ρintracellular were determined from the impedance data. Glycogen content (GC) was visualized histologically and quantified biochemically. At frequencies > 1 MHz, Pompe mice demonstrated significantly decreased longitudinal and transverse conductivity, increased Cole-Cole parameters, ρo and ρo-ρ∞, and decreased ρintracellular. Changes in longitudinal conductivity and ρintracellular correlated with increased GC in Pompe animals. Ex vivo high frequency impedance measures are sensitive to alterations in intracellular myofiber features considered characteristic of GSDII, making them potentially useful measures of disease status.

Application of minimally invasive methods to enable the measurement of tissue permittivity in the neuromuscular clinic remain elusive. This paper provides a theoretical and modeling study on the measurement of the permittivity of two-dimensional anisotropic tissues such as skeletal muscle with a multi-electrode cross-shaped needle. For this, we design a novel cross-shaped needle with multiple-electrodes and analyse apparent impedance corresponding to the measured impedance. In addition, we propose three methods of estimate anisotropic muscle permittivity. Compared to existing electrical impedance-based needle methods that we have developed, the new needle design and numerical methods associated enable estimating in vivo muscle permittivity values with only a single needle insertion. Being able to measure muscle permittivity directly with a single needle insertion could open up an entirely new area of research with direct clinical application, including using these values to assist in neuromuscular diagnosis and to assess subtle effects of therapeutic intervention on muscle health.

Canine degenerative myelopathy (DM) leads to disuse and neurogenic muscle atrophy. Currently there is a lack of non-invasive quantitative measures of muscle health in dogs with DM. Muscle pathology has been previously quantified in other disorders using the technique of electrical impedance myography (EIM) but it has not been reported for DM. The objective of this study was to compare EIM between DM-affected and similar aged healthy dogs as well as assess EIM changes over time in DM-affected dogs. Multifrequency EIM was performed on DM affected dogs at baseline and during disease progression and on age-matched healthy dogs. Muscles evaluated in the pelvic limbs included the craniotibialis, gastrocnemius, gracilis, sartorius, and biceps femoris. The 100 kHz phase angle was extracted from the full frequency set for analysis. Phase values were lower in DM dogs as compared to healthy controls. Specifically, phase of the gastrocnemius was lower on the left (θ = 7.69, 13.06; p =0.002) and right (θ= 6.11, 11.72; p = 0.001) in DM vs. control dogs, respectively. The mean phase value of all measured muscles was also lower on the left (θ = 9.24, 11.62; p = 0.012) and right (θ = 9.18, 11.72; p = 0.021). Other individual muscles measured did not reach statistical significance, although values were consistently lower in DM-affected dogs. With disease progression, downward trends in phase values were detected in DM-affected dogs when monitored serially over time. This study demonstrates that EIM 100 kHz phase values are sensitive to muscle pathology in DM and that phase values are decreased in dogs with DM. Measurements from the gastrocnemius muscle show the greatest differences from similar aged healthy dogs suggesting it may be the preferred muscle for future EIM studies.

INTRODUCTION/AIMS: We assessed the classification performance of machine learning (ML) using multifrequency electrical impedance myography (EIM) values to improve upon diagnostic outcomes as compared to those based on a single EIM value.

METHODS: EIM data was obtained from unilateral excised gastrocnemius in eighty diseased mice (26 D2-mdx, Duchenne muscular dystrophy model, 39 SOD1G93A ALS model, and 15 db/db, a model of obesity-induced muscle atrophy) and 33 wild-type (WT) animals. We assessed the classification performance of a ML random forest algorithm incorporating all the data (multifrequency resistance, reactance and phase values) comparing it to the 50 kHz phase value alone.

RESULTS: ML outperformed the 50 kHz analysis as based on receiver-operating characteristic curves and measurement of the area under the curve (AUC). For example, comparing all diseases together versus WT from the test set outputs, the AUC was 0.52 for 50 kHz phase, but was 0.94 for the ML model. Similarly, when comparing ALS versus WT, the AUCs were 0.79 for 50 kHz phase and 0.99 for ML.

DISCUSSION: Multifrequency EIM using ML improves upon classification compared to that achieved with a single-frequency value. ML approaches should be considered in all future basic and clinical diagnostic applications of EIM.

The purpose of this study was to explore the effect an individual's exercise training type will have on muscle fatigability during repetitive contractions induced by Neuromuscular Electrical Stimulation (NMES). Thirty-four subjects comprising of competitive athletes and controls were recruited into three cohorts: Endurance (runners/cyclists) n = 13; nine male, four female; 27 ± 8 years old, Explosive (Lifters/Sprinters) n = 11; nine male, two female; 30 ± 7 years old, and controls n = 10, six male, four female, 26 ± 4 years old. Subjects were placed in a custom-made leg extension rig, and received NMES against a fixed resistance (NMES-FR), to the Vastus Medialis muscle resulting in isometric leg extensions, at a duty cycle of 1 s on/3 s rest, for 20 min. The force of the isometric contractions was recorded using a Hogan MicroFet2 dynamometer, and three separate fatigue metrics were calculated to compare the different cohorts, sports within each cohort, and gender within each cohort. For every fatigue metric, the endurance group fatigued significantly less than both the explosive and control cohorts, with no difference observed between the explosive and the controls. Within each cohort, no significant difference was observed in any fatigue metric between sport or gender, but these comparisons lacked power. The results show that only high capacity endurance activity will have any effect on reducing one's fatigability during repetitive NMES. The implications of this conclusion can aid in the development of NMES regimens for use in healthy populations, such as athletic training or astronaut musculoskeletal countermeasures, as well as clinical applications when fatigue is to be minimized.

Electrical impedance myography (EIM) is a convenient technique that can be used in preclinical and clinical studies to assess muscle tissue health and disease. EIM is obtained by applying a low-intensity, directionally focused, electrical current to a muscle of interest across a range of frequencies (i.e., from 1 kHz to 10 MHz) and recording the resulting voltages. From these, several standard impedance components, including the reactance, resistance, and phase, are obtained. When performing ex vivo measurements on excised muscle, the inherent passive electrical properties of the tissue, namely the conductivity and relative permittivity, can also be calculated. EIM has been used extensively in animals and humans to diagnose and track muscle alterations in a variety of diseases, in relation to simple disuse atrophy, or as a measure of therapeutic intervention. Clinically, EIM offers the potential to track disease progression over time and to assess the impact of therapeutic interventions, thus offering the opportunity to shorten the clinical trial duration and reduce sample size requirements. Because it can be performed noninvasively or minimally invasively in living animal models as well as humans, EIM offers the potential to serve as a novel translational tool enabling both preclinical and clinical development. This article provides step-by-step instructions on how to perform in vivo and ex vivo EIM measurements in mice and rats, including approaches to adapt the techniques to specific conditions, such as for use in pups or obese animals.