Electrical Impedance Myography in Dogs With Degenerative Myelopathy.

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.

Electrical impedance myography in healthy dogs: Normative values, repeatability, and the impact of age.

Convenient tools to assess canine skeletal muscle health would be useful for a variety of applications, including standard veterinary assessments of dog fitness, as well as studies of muscle deterioration due to age or disease. One technology that can be applied conveniently to awake dogs with minimal restraint is electrical impedance myography (EIM). In EIM, a weak electrical current is applied via surface electrodes to a muscle of interest and consequent impedance characteristics of the muscle are obtained, providing insight into muscle condition and composition. In this study, we assessed a total of 73 dogs (42 males and 31 females), of varied neutering status and breed, ages 0.6 to 13.5 years. We identified age-dependent reference values for the 100 kHz phase value in three pelvic limb muscles, caudal sartorius, cranial tibial, and gastrocnemius. While phase values were generally higher in males than females, the difference did not reach significance. In general, values declined on average with age at about 0.5 degrees/year, but with the decline being most substantial in the oldest dogs. Limited reproducibility assessment of the technique suggested good repeatability with variation in values between measurements being under 5%. These results show that EIM has the potential for the assessment of canine muscle health and may find value in aging muscle research.

Brain diffusion tensor imaging in dogs with degenerative myelopathy.

BACKGROUND: Degenerative myelopathy (DM) in dogs shares similarities with superoxide dismutase 1-associated human amyotrophic lateral sclerosis (ALS). Brain microstructural lesions are quantified using diffusion tensor imaging (DTI) in ALS patients. OBJECTIVE: Characterize brain neurodegenerative changes in DM-affected dogs using DTI. ANIMALS: Sixteen DM-affected and 8 control dogs. METHODS: Prospective observational study. Brain DTI was performed at baseline and every 3 months on DM-affected dogs and compared to controls. Fractional anisotropy, mean diffusivity, axial diffusivity, and radial diffusivity were calculated on specified regions of interest. Gait scores (0, normal to 14, tetraplegia) were assigned at each scan. Diffusion tensor imaging values in DM-affected dogs were compared to controls, gait scores, and evaluated over time. RESULTS: Mean age was 5.7 years (SD 3.2) in controls and 9.7 years (SD 1.4) in DM-affected dogs. In DM-affected dogs, mean baseline gait score was 4 (SD 1), and mean score change from baseline to last scan was 4.82 (SD 2.67). Nine dogs had ≤3 scans; 7 had >3 scans. Accounting for age, no differences in DTI indices were identified for any brain or proximal spinal cord regions between DM-affected dogs and controls (P > .05). Diffusion tensor imaging values poorly correlated with gait scores (R2 < .2). No significant changes were identified in diffusion indices over time (P > .05). CONCLUSIONS AND CLINICAL IMPORTANCE: Diffusion tensor imaging indices did not differentiate DM-affected from control dogs, detect longitudinal changes, or differentiate disease severity. Findings do not yet support brain DTI as an imaging biomarker.

Intravitreal enzyme replacement inhibits progression of retinal degeneration in canine CLN2 neuronal ceroid lipofuscinosis.

CLN2 neuronal ceroid lipofuscinosis is a rare recessive hereditary retinal and neurodegenerative disease resulting from deleterious sequence variants in TPP1 that encodes the soluble lysosomal enzyme tripeptidyl peptidase-1 (TPP1). Children with this disorder develop normally, but starting at 2-4 years of age begin to exhibit neurological signs and visual deficits. Vision loss that progresses to blindness is associated with progressive retinal degeneration and impairment of retinal function. Similar progressive loss of retinal function and retinal degeneration occur in a dog CLN2 disease model with a TPP1 null sequence variant. Studies using the dog model were conducted to determine whether intravitreal injection of recombinant human TPP1 (rhTPP1) administered starting after onset of retinal functional impairment could slow or halt the progression of retinal functional decline and degeneration. TPP1-null dogs received intravitreal injections of rhTPP1 in one eye and vehicle in the other eye beginning at 23.5-25 weeks of age followed by second injections at 34-40 weeks in 3 out of 4 dogs. Ophthalmic exams, in vivo ocular imaging, and electroretinography (ERG) were repeated regularly to monitor retinal structure and function. Retinal histology was evaluated in eyes collected from these dogs when they were euthanized at end-stage neurological disease (40-45 weeks of age). Intravitreal rhTPP1 injections were effective in preserving retinal function (as measured with the electroretinogram) and retinal morphology for as long as 4 months after a single treatment. These findings indicate that intravitreal injection of rhTPP1 administered after partial loss of retinal function is an effective treatment for preserving retinal structure and function in canine CLN2 disease.

Isolation and preliminary characterization of amino acid substitution mutations that increase the activity of the osmoregulated ProP protein of Salmonella enterica serovar Typhimurium.

In Enterobacteriaceae, the ProP protein, which takes up proline and glycine betaine, is subject to a post-translational control mechanism that increases its activity at high osmolarity. In order to investigate the osmoregulatory mechanism of the Salmonella enterica ProP, we devised a positive selection for mutations that conferred increased activity on this protein at low osmolarity. The selection involved the isolation of mutations in a proline auxotroph that resulted in increased accumulation of proline via the ProP system in the presence of glycine betaine, which is a competitive inhibitor of proline uptake by this permease. This selection was performed by first-year undergraduates in two semesters of a research-based laboratory course. The students generated sixteen mutations resulting in six different single amino acids substitutions. They determined the effects of the mutations on the growth rates of the cells in media of high and low osmolarity in the presence of low concentrations of proline or glycine betaine. Furthermore, they identified the mutations by DNA sequencing and displayed the mutated amino acids on a putative three-dimensional structure of the protein. This analysis suggested that all six amino acid substitutions are residues in trans-membrane helices that have been proposed to contribute to the formation of the transport pore, and, thus, may affect the substrate binding site of the protein.