Evaluation of the diagnostic value of transcranial electrical stimulation (TES) to assess neuronal functional integrity in horses.

Medical imaging allows for the visualization of spinal cord compression sites; however, it is impossible to assess the impact of visible stenotic sites on neuronal functioning, which is crucial information to formulate a correct prognosis and install targeted therapy. It is hypothesized that with the transcranial electrical stimulation (TES) technique, neurological impairment can be reliably diagnosed. Objective: To evaluate the ability of the TES technique to assess neuronal functional integrity in ataxic horses by recording TES-induced muscular evoked potentials (MEPs) in three different muscles and to structurally involve multiple ancillary diagnostic techniques, such as clinical neurological examination, plain radiography (RX) with ratio assessment, contrast myelography, and post-mortem gross and histopathological examination. Methods: Nine ataxic horses, showing combined fore and hindlimb ataxia (grades 2-4), were involved, together with 12 healthy horses. TES-induced MEPs were recorded bilaterally at the level of the trapezius (TR), the extensor carpi radialis (ECR), and tibialis cranialis (TC) muscles. Two Board-certified radiologists evaluated intra- and inter-sagittal diameter ratios on RX, reductions of dorsal contrast columns, and dural diameters (range skull-T1). Post-mortem gross pathological and segmental histopathological examination was also performed by a Board-certified pathologist. Results: TES-MEP latencies were significantly prolonged in both ECR and TC in all ataxic horses as opposed to the healthy horses. The TR showed a mixed pattern of normal and prolonged latency times. TES-MEP amplitudes were the least discriminative between healthy and ataxic horses. Youden's cutoff latencies for ataxic horses were 24.6 ms for the ECR and 45.5 ms for the TC (sensitivity and specificity of 100%). For healthy horses, maximum latency values were 22 and 37 ms, respectively. RX revealed spinal cord compression in 8 out of 9 involved ataxic horses with positive predictive values of 0-100%. All ataxic horses showed multi-segmental Wallerian degeneration. All pathological changes recorded in the white matter of the spinal cord were widely dispersed across all cervical segments, whereas gray matter damage was more localized at the specific segmental level. Conclusion: TES-MEP latencies are highly sensitive to detect impairment of spinal cord motor functions for mild-to-severe ataxia (grades 2-4).

Presumed residual thymic tissue is a common finding in thoracic computed tomography in adult dogs.

Residual thymic tissue is a common incidental finding in thoracic CT of human adults. To determine whether presumed residual thymic tissue is also a common incidental finding in adult dogs, a two part-study was performed. The first part was a prospective, descriptive design where CT examination was performed in six canine cadavers within 24 h after death and presumed residual thymic tissue was examined pathologically. The second part of the study was a retrospective, analytical design where medical records of our institution were searched for thoracic CT scans of adult dogs performed in the year 2020. Age, sex, breed, presence of presumed thymic tissue, location, shape, attenuation, homogeneity, and width of the tissue were recorded and comparisons were performed using these data. In 4 of 6 of the prospective cases, thymic tissue was present on histology and in 2 of 6 dogs the presence of thymic tissue could not be confirmed. For the retrospective study, in 161/169 (95.3%) cases with presumed residual thymic tissue were detected. Shape and size were highly variable with either homogeneous (46.6%), heterogeneous (42.9%), or mixed (10.6%) attenuation. Dogs with presumed residual thymic tissue were significantly younger (median: 9.1 years; range: 1.2-14.3 vs. median: 10.5 years; range: 9.4-12.3) as were dogs with homogeneous attenuation of the tissue (median: 8.1 years; range: 1.2-14.3 vs. median: 9.5 years; range: 4.0-14.3). In conclusion, results indicated that presumed residual thymic tissue is a common CT finding in adult dogs and can be considered incidental.

Histological and magnetic resonance imaging assessment of Liqoseal in a spinal in vivo pig model.

BACKGROUND: Liqoseal (Polyganics, B.V.) is a dural sealant patch for preventing postoperative cerebrospinal fluid (CSF) leakage. It has been extensively tested preclinically and CE (Conformité Européenne) approved for human use after a first cranial in-human study. However, the safety of Liqoseal for spinal application is still unknown. The aim of this study was to assess the safety of spinal Liqoseal application compared with cranial application using histology and magnetic resonance imaging characteristics. METHODS: Eight female Dutch Landrace pigs underwent laminectomy, durotomy with standard suturing and Liqoseal application. Three control animals underwent the same procedure without sealant application. The histological characteristics and imaging characteristics of animals with similar survival times were compared to data from a previous cranial porcine model. RESULTS: Similar foreign body reactions were observed in spinal and cranial dura. The foreign body reaction consisted of neutrophils and reactive fibroblasts in the first 3 days, changing to a chronic granulomatous inflammatory reaction with an increasing number of macrophages and lymphocytes and the formation of a fibroblast layer on the dura by day 7. Mean Liqoseal plus dura thickness reached a maximum of 1.2 mm (range 0.7-2.0 mm) at day 7. CONCLUSION: The spinal dural histological reaction to Liqoseal during the first 7 days was similar to the cranial dural reaction. Liqoseal did not swell significantly in both application areas over time. Given the current lack of a safe and effective dural sealant for spinal application, we propose that an in-human safety study of Liqoseal is the logical next step.

Trapezius Motor Evoked Potentials From Transcranial Electrical Stimulation and Transcranial Magnetic Stimulation: Reference Data, Characteristic Differences and Intradural Motor Velocities in Horses.

Reason for Performing Study: So far, only transcranial motor evoked potentials (MEP) of the extensor carpi radialis and tibialis cranialis have been documented for diagnostic evaluation in horses. These allow for differentiating whether lesions are located in either the thoraco-lumbar region or in the cervical myelum and/or brain. Transcranial trapezius MEPs further enable to distinguish between spinal and supraspinal located lesions. No normative data are available. It is unclear whether transcranial electrical stimulation (TES) and transcranial magnetic stimulation (TMS) are interchangeable modalities. Objectives: To provide normative data for trapezius MEP parameters in horses for TES and TMS and to discern direct and indirect conduction routes by neurophysiological models that use anatomical geometric characteristics to relate latency times with peripheral (PCV) and central conduction velocities (CCV). Methods: Transcranial electrical stimulation-induced trapezius MEPs were obtained from twelve horses. TES and TMS-MEPs (subgroup 5 horses) were compared intra-individually. Trapezius MEPs were measured bilaterally twice at 5 intensity steps. Motoneurons were localized using nerve conduction models of the cervical and spinal accessory nerves (SAN). Predicted CCVs were verified by multifidus MEP data from two horses referred for neurophysiological assessment. Results: Mean MEP latencies revealed for TES: 13.5 (11.1-16.0)ms and TMS: 19.7 (12-29.5)ms, comprising ∼100% direct routes and for TMS mixed direct/indirect routes of L:23/50; R:14/50. Left/right latency decreases over 10 > 50 V for TES were: -1.4/-1.8 ms and over 10 > 50% for TMS: -1.7/-3.5 ms. Direct route TMS-TES latency differences were 1.88-4.30 ms. 95% MEP amplitudes ranges for TES were: L:0.26-22 mV; R:0.5-15 mV and TMS: L:0.9 - 9.1 mV; R:1.1-7.9 mV. Conclusion: This is the first study to report normative data characterizing TES and TMS induced- trapezius MEPs in horses. The complex trapezius innervation leaves TES as the only reliable stimulation modality. Differences in latency times along the SAN route permit for estimation of the location of active motoneurons, which is of importance for clinical diagnostic purpose. SAN route lengths and latency times are governed by anatomical locations of motoneurons across C2-C5 segments. TES intensity-dependent reductions of trapezius MEP latencies are similar to limb muscles while MEP amplitudes between sides and between TES and TMS are not different. CCVs may reach 180 m/s.

Intervertebral disc degeneration in warmblood horses: Histological and biochemical characterization.

Gross morphology of healthy and degenerated intervertebral discs (IVDs) is largely similar in horses as in dogs and humans. For further comparison, the biochemical composition and the histological and biochemical changes with age and degeneration were analyzed in 41 warmblood horses. From 33 horses, 139 discs and 2 fetal vertebral columns were evaluated and scored histologically. From 13 horses, 73 IVDs were assessed for hydration, DNA, glycosaminoglycans, total collagen, hydroxyl-lysyl-pyridinoline, hydroxylysine, and advanced glycation end-product (AGE) content. From 7 horses, 20 discs were assessed for aggrecan, fibronectin, and collagen type 1 and 2 content. Histologically, tearing of the nucleus pulposus (NP) and cervical annulus fibrosus (AF), and total histological score (tearing and vascular proliferation of the AF, and chondroid metaplasia, chondrocyte-like cell proliferation, presence of notochordal cells, matrix staining, and tearing of the NP) correlated with gross degeneration. Notochordal cells were not seen in IVDs of horses. Age and gross degeneration were positively correlated with AGEs and a fibrotic phenotype, explaining gross degenerative changes. In contrast to dogs and humans, there was no consistent difference in glycosaminoglycan content and hydration between AF and NP, nor decrease of these variables with age or degeneration. Hydroxylysine decrease and collagen 1 and AGEs increase were most prominent in the NP, suggesting degeneration started in the AP. In caudal cervical NPs, AGE deposition was significantly increased in grossly normal IVDs and total collagen significantly increased with age, suggesting increased biomechanical stress and likelihood for spinal disease in this part of the vertebral column.

Safety and biodegradability of a synthetic dural sealant patch (Liqoseal) in a porcine cranial model.

Background: Liqoseal consists of a watertight layer of poly(ester)ether urethane and an adhesive layer containing polyethylene glycol-N-hydroxysuccinimide (PEG-NHS). It is designed to prevent cerebrospinal fluid (CSF) leakage after intradural surgery. This study assessed the safety and biodegradability of Liqoseal in a porcine craniotomy model. Methods: In 32 pigs a craniotomy plus durotomy was performed. In 15 pigs Liqoseal was implanted, in 11 control pigs no sealant was implanted and in 6 control pigs a control dural sealant (Duraseal or Tachosil) was implanted. The safety of Liqoseal was evaluated by clinical, MRI and histological assessment. The degradation of Liqoseal was histologically estimated. Results: Liqoseal, 2 mm thick before application, did not swell and significantly was at maximum mean thickness of 2.14 (±0.37) mm at one month. The foreign body reaction induced by Liqoseal, Duraseal and Tachosil were comparable. Liqoseal showed no adherence to the arachnoid layer and was completely resorbed between 6 and 12 months postoperatively. In one animal with Liqoseal, an epidural fluid collection containing CSF could not be excluded. Conclusion: Liqoseal seems to be safe for intracranial use and is biodegradable. The safety and performance in humans needs to be further assessed in clinical trials.

Cervical articular process joint osteochondrosis in Warmblood foals.

BACKGROUND: In Warmblood horses, degenerative joint disease is involved in cervical malformation and malarticulation (CVM). The degree of contribution of articular process joint (APJ) osteochondrosis (OC) is not clear. OBJECTIVES: (a) To explore the presence of predilection sites for APJ OC in cervical and cranial thoracic vertebral columns of Warmblood foals and (b) to examine the correlation of such a site with the predilection site of CVM. STUDY DESIGN: Case series. METHODS: Seven hundred APJ facets of C2 to T2 of 29 foals (11 months gestation to 12 months [median age 7 days; range 365 days; 95% confidence interval [95% CI] 2-47 days]) were examined for OC and prevalence between joints, and the predilection site for CVM and the cranial cervical vertebral column were evaluated. RESULTS: About 20.6% of facets revealed OC. There was no predilection site. Prevalence decreased with age up to 1 year (odds ratio [OR] 0.997; (95% CI 0.975-0.998)) but not up to 5 months. Severity increased with age in all age ranges (up to 1 year OR 1.023; 95% CI 1.005-1.049; >1-5 months, OR 1.203; 95% CI 1.014e+00-1.921; up to 1 month, OR 1.114; 95% CI 1.041-1.228). Highest prevalence was in cranial facets of the cervical and cervical-thoracic joints and in caudal facets of the thoracic joint up to 1 year and up to 1 month (OR 0.364; 95% CI 0.170-0.745, OR 0.434; 95% CI: 0.235-0.782, OR 7.665; 95% CI: 1.615-66.553 and OR 0.400; 95% CI 0.170-0.880, OR 0.351; 95% CI 0.172-0.700, OR 5.317; 95% CI 1.098-44.344 respectively). MAIN LIMITATIONS: Two-thirds of the foals were less than 1 month of age. CONCLUSIONS: Articular process joint OC in Warmblood foals is common and is not more prevalent at CVM predilection sites, suggesting that abnormalities of enchondral ossification may not be major contributors to CVM.

Histologic Comparison of the Dura Mater among Species.

The biocompatibility, biodegradation, feasibility, and efficacy of medical devices like dural sealants and substitutes are often evaluated in various animal models. However, none of these studies explain the rationale for choosing a particular species, and a systematic interspecies comparison of the dura is not available. We hypothesized that histologic characteristics of the dura would differ among species. We systematically investigated basic characteristics of the dura, including thickness, composition, and fibroblast orientation of the dura mater, in 34 samples representing 10 animal species and compared these features with human dura by using hematoxylin and eosin staining and light microscopy. Dura showed many similarities between species in terms of composition. In all species, dura consisted of at least one fibrovascular layer, which contained collagen, fibroblasts, and blood vessels, and a dural border cell layer beneath the fibrovascular layer. Differences between species included the number of fibrovascular layers, fibroblast orientation, and dural thickness. Human dura was the thickest (564 µm) followed by equine (313 µm), bovine (311 µm), and porcine (304 µm) dura. Given the results of this study and factors such as gross anatomy, feasibility, housing, and ethical considerations, we recommend the use of a porcine model for dural research, especially for in vivo studies.

State-of-the-Art Diagnostic Methods to Diagnose Equine Spinal Disorders, With Special Reference to Transcranial Magnetic Stimulation and Transcranial Electrical Stimulation.

Spinal cord disorders are a common problem in equine medicine. However, finding the site of the lesion is challenging for veterinarians because of a lack of sensitive diagnostic methods that can assess neuronal functional integrity in horses. Although medical imaging is frequently applied to help diagnose corticospinal disorders, this approach does not reveal functional information. For the latter, transcranial magnetic stimulation (TMS) and more recently transcranial electrical stimulation (TES) can be useful. These are brain stimulation techniques that create either magnetic or electrical fields passing through the motor cortex, inducing muscular responses, which can be recorded either intramuscularly or extramuscularly by needle or surface electrodes. This permits the evaluation of the functional integrity of the spinal motor tracts and the nerve conduction pathways. The interest in TES in human medicine emerged these last years because unlike TMS, TES tends to bypass the motor cortex of the brain and predominantly relies on direct activation of corticospinal and extrapyramidal axons. Results from human medicine have indicated that TMS and TES recordings are mildly if not at all affected by sedation. Therefore, this technique can be reliably used in human patients under either sedation or full anesthesia to assess functional integrity of the corticospinal and adjunct motor tracts. This opens important new avenues in equine medicine.

Equine cervical intervertebral disc degeneration is associated with location and MRI features.

Morphology of the equine cervical intervertebral disc is different from that in humans and small companion animals and published imaging data are scarcely available. The objectives of this exploratory, methods comparison study were (a) to describe MRI features of macroscopically nondegenerated and degenerated intervertebral discs (b) to test associations between spinal location and macroscopic degeneration or MRI-detected annular protrusion and between MRI-detected annular protrusion and macroscopic degeneration, and (c) to define MRI sequences for characterizing equine cervical intervertebral disc degeneration. Ex vivo MRI of intervertebral discs was performed in 11 horses with clinical signs related to the cervical region prior to macroscopic assessment. Mixed-effect logistic regression modeling included spinal location, MRI-detected annular protrusion, and presence of macroscopic degeneration with "horse" as random effect. Odds ratio and 95% confidence interval were determined. Reduced signal intensity in proton density turbo SE represented intervertebral disc degeneration. Signal voids due to presence of gas and/or hemorrhage were seen in gradient echo sequences. Presence of macroscopic intervertebral disc degeneration was significantly associated with spinal location with odds being higher in the caudal (C5 to T1) versus cranial (C2 to C5) part of the cervical vertebral column. Intervertebral discs with MRI-detected annular protrusion grades 2-4 did have higher odds than with grade 1 to have macroscopic degeneration. It was concluded that MRI findings corresponded well with gross macroscopic data. Magnetic resonance imaging of the equine cervical intervertebral disc seems to be a promising technique, but its potential clinical value for live horses needs to be explored further in a larger and more diverse population of horses.

Magnetic resonance imaging appearance of the brain and cervical spinal cord in an edema disease affected pig.

A 7-week-old male pig was presented with signs of a central nervous system disorder. An MRI of the head and cervical spine was performed immediately after euthanasia. The MRI revealed multifocal bilaterally symmetric T2-weighted hyperintense lesions in the brain and spinal cord, likely due to a toxic metabolic process. Histopathological examination supported the MRI findings and confirmed the diagnosis of edema disease due to Shiga-like toxin produced by Escherichia coli. This is the first case published of the MRI findings in an edema disease affected pig.

Incomplete endothelialization of an intravascular implant and fatal late-onset bacterial ductal arteritis in a dog with occluded patent ductus arteriosus.

An 18-month-old male Akita Inu dog developed fever and lameness 8 months after successful transcatheter closure of a patent ductus arteriosus with an Amplatz Canine Duct Occluder (ACDO). Corynebacterium species were cultured from 3 blood samples. Echocardiography showed a vegetative process on the aortic valves. The dog died spontaneously 3 days after development of the initial signs. Necropsy confirmed the presence of bacterial ductal arteritis and myocarditis, and revealed an incomplete endothelialization of the intraductal metal implant. The reason for the lack of (neo)endothelialization of the ACDO remains unknown. We conclude that late-onset bacterial device-related ductal arteritis can develop in dogs where the implant is incompletely covered by a protective endothelial layer.

Intervertebral Disc Degeneration in Warmblood Horses: Morphology, Grading, and Distribution of Lesions.

Equine intervertebral disc degeneration is thought to be rare and of limited clinical relevance, although research is lacking. To objectively assess pathological changes of the equine intervertebral disc and their clinical relevance, description of the normal morphology and a practical, biologically credible grading scheme are needed. The objectives of this study are to describe the gross and histological appearance of the equine intervertebral discs and to propose a grading scheme for macroscopic degeneration. Spinal units from 33 warmblood horses were grossly analyzed and scored. Of the 286 intervertebral discs analyzed, 107 (37%) were assigned grade 1 and grade 2 (considered normal) and were analyzed histologically. A nucleus pulposus and an annulus fibrosus could be identified macroscopically and histologically. Histologically, the nucleus pulposus was composed of a cartilaginous matrix and the annulus fibrosus of parallel collagenous bands. A transition zone was also histologically visible. Intra- and inter-observer reliability scores were high for all observers. Higher grades were associated with greater age. Gross changes associated with equine intervertebral disc degeneration (grades 3-5)-that is, yellow discoloration, cleft formation (tearing), and changes in consistency of the nucleus pulposus-were largely similar to those in humans and dogs and were most prevalent in the caudal cervical spine. Equine intervertebral disc degeneration was not associated with osteophyte formation. Changes of the vertebral bone were most common in the thoracolumbar spine but were not correlated with higher grades of intervertebral disc degeneration. Thus, changes of the vertebral bone should be excluded from grading for equine intervertebral disc degeneration.

Usefulness of Sealants for Dural Closure: Evaluation in an In Vitro Model.

BACKGROUND: Cerebrospinal fluid (CSF) leakage occurs in 4% to 32% of cranial surgeries and is associated with significant patient burden and expense. The use of sealant as an adjunct to primary dural closure is assumed to help prevent CSF leakage. OBJECTIVE: To examine the utility of different sealants for dural closure using an in Vitro model. METHODS: We evaluated 9 commonly used dural sealants, including Tachosil (Takeda Inc, Osaka, Japan), Adherus (Hyperbranch Inc, Durham, North Carolina), Duraform (Codman, Raynham, Massachusetts), Tissudura (Baxter, Deerfield, Illinois), Hemopatch (Baxter), TissuePatchDural (Tissuemed, Leeds, United Kingdom), Tisseel (Baxter), Duragen Secure (Integra, Plainsboro, New Jersey), and Duraseal, (Integra). Sealants were tested in 2 novel in Vitro setups using fresh porcine dura: the first tested the acute burst pressure of a sealed 3-mm gap, while the second examined resistance to a pressure wave mimicking intracranial pressure for 72 h. RESULTS: Adherus showed the highest mean burst pressure (87 ± 47 mmHg) followed by Tachosil (71 ± 16 mmHg) and Duraseal (51 ± 42 mmHg); these were the only 3 sealants showing burst pressures above normal physiological intracranial pressure. In the 72-h setup, only Adherus and Duraseal maintained appropriate sealing for the duration of the experiment. Tachosil released from the dura after 1.4 h (95% confidence interval, -1.8-4.7). CONCLUSION: Given the high cost of sealants and the results of this study, we advocate a critical attitude toward sealant application as an adjunct to classic dural closure.

Effects of long-term use of the preferential COX-2 inhibitor meloxicam on growing pigs.

Meloxicam, a preferential COX-2 inhibitor, is a commonly used NSAID in pigs. Besides having potential side effects on the gastrointestinal tract, this type of drug might potentially affect osteogenesis and chondrogenesis, processes relevant to growing pigs. Therefore, the effects of long-term meloxicam treatment on growing pigs were studied. Twelve piglets (n=6 receiving daily meloxicam 0.4 mg/kg orally from 48 until 110 days of age; n=6 receiving only applesauce (vehicle control)) were subjected to visual and objective gait analysis by pressure plate measurements at several time points. Following euthanasia a complete postmortem examination was performed and samples of the talus and distal tibia, including the distal physis, were collected. Trabecular bone microarchitecture was analysed by microCT scanning, bone stiffness by compression testing and growth plate morphology using light microscopy. Animals were not lame and gait patterns did not differ between the groups. Pathological examination revealed no lesions compatible with known side effects of NSAIDs. Trabecular bone microarchitecture and growth plate morphology did not differ between the two groups. The findings of this in vivo study reduce concerns regarding the long-term use of meloxicam in young, growing piglets.

EX VIVO COMPUTED TOMOGRAPHIC EVALUATION OF MORPHOLOGY VARIATIONS IN EQUINE CERVICAL VERTEBRAE.

Diagnostic imaging is one of the pillars in the clinical workup of horses with clinical signs of cervical spinal disease. An improved awareness of morphologic variations in equine cervical vertebrae would be helpful for interpreting findings. The aim of this anatomic study was to describe CT variations in left-right symmetry and morphology of the cervical and cervicothoracic vertebrae in a sample of horses. Postmortem CT examinations of the cervical spine for horses without congenital growth disorders were prospectively and retrospectively recruited. A total of 78 horses (27 foals, 51 mature horses) were evaluated. Twenty-six horses (33.3%) had homologous changes in which a transposition of the caudal part of the transverse process (caudal ventral tubercle) of C6 toward the ventral aspect of the transverse process of C7 was present (n = 10 bilateral, n = 12 unilateral left-sided, n = 4 unilateral right-sided). There was one horse with occipito-atlantal malformation, two horses with rudimentary first ribs bilaterally, and one horse with bilateral transverse processes at Th1, representing homeotic (transitional) vertebral changes. Chi-square tests identified no significant differences in the number of conformational variations between the group of mature horses with or without clinical signs (P = 0.81) or between the group of mature horses and the group of foals (P = 0.72). Findings indicated that, in this sample of horses, the most frequently identified variations were homologous variations (transposition of the caudal part of the transverse process of C6-C7) in the caudal equine cervical vertebral column. Homeotic (transitional) variations at the cervicothoracic vertebral column were less common.

Left ventricular cardiac myxoma and sudden death in a dog.

BACKGROUND: Myxoma is a very rare benign cardiac tumor in dogs. This is the first description of a cardiac myxoma originating from the left ventricular outflow tract, presumably causing sudden death. CASE PRESENTATION: A previously healthy 12-year-old male West Highland white terrier was found dead during its 1-week stay in a kennel. The dog was known to have a cardiac murmur. On necropsy, a pedunculated neoplasia was found attached to the interventricular aspect of the left ventricular outflow tract, resulting in almost complete obstruction of the aorta. As this was the only abnormality identified, the tumor was considered as the cause of sudden death. Histopathologic findings were compatible with a myxoma. CONCLUSIONS: Benign intraluminal tumors of the heart are very rare in dogs, but may have fatal consequences. Echocardiography could have revealed the cause of the cardiac murmur of this previously asymptomatic dog. Surgical removal could have been possible, as the tumor was pedunculated.