Neutrophil extracellular traps in CSF and serum of dogs with steroid-responsive meningitis-arteritis.

In steroid-responsive meningitis-arteritis (SRMA), inflammatory dysregulation is driven by neutrophilic granulocytes resulting in purulent leptomeningitis. Neutrophils can generate neutrophil extracellular traps (NET). Uncontrolled NET-formation or impaired NET-clearance evidently cause tissue and organ damage resulting in immune-mediated diseases. The aim of the study was to verify that NET-formation is detectable in ex vivo samples of acute diseased dogs with SRMA by visualizing and measuring NET-markers in serum and cerebrospinal fluid (CSF) samples. CSF-samples of dogs with acute SRMA (n = 5) and in remission (n = 4) were examined using immunofluorescence (IF)-staining of DNA-histone-1-complexes, myeloperoxidase and citrullinated Histone H3 (H3Cit). Immunogold-labeling of H3Cit and neutrophil elastase followed by transmission electron microscopy (TEM) were used to determine ultrastructural NET-formation in the CSF of one exemplary dog. H3Cit-levels and DNase-activity were measured in CSF and serum samples using an H3Cit-ELISA and a DNase-activity-assay, respectively in patients with the following diseases: acute SRMA (n = 34), SRMA in remission (n = 4), bacterial encephalitis (n = 3), meningioma with neutrophilic inflammation (n = 4), healthy dogs (n = 6). NET-formation was detectable with IF-staining in n = 3/5 CSF samples of dogs with acute SRMA but were not detectable during remission. Vesicular NET-formation was detectable in one exemplary dog using TEM. DNase-activity was significantly reduced in dogs suffering from acute SRMA compared to healthy control group (p < 0.0001). There were no statistical differences of H3Cit levels in CSF or serum samples of acute diseased dogs compared to dogs under treatment, dogs suffering from meningioma or bacterial encephalitis or the healthy control group. Our findings demonstrate that NET-formation and insufficient NET-clearance possibly drive the immunologic dysregulation and complement the pathogenesis of SRMA. The detection of NETs in SRMA offers many possibilities to explore the aetiopathogenetic influence of this defence mechanism of the innate immune system in infectious and non-infectious canine neuropathies.

SLC19A3 Loss-of-Function Variant in Yorkshire Terriers with Leigh-Like Subacute Necrotizing Encephalopathy.

Sporadic occurrence of juvenile-onset necrotizing encephalopathy (SNE) has been previously reported in Yorkshire terriers. However, so far, no causative genetic variant has been found for this breed-specific form of suspected mitochondrial encephalomyopathy. Affected dogs showed gait abnormalities, central visual defects, and/or seizures. Histopathological analysis revealed the presence of major characteristics of human Leigh syndrome and SNE in Alaskan huskies. The aim of this study was to characterize the genetic etiology of SNE-affected purebred Yorkshire terriers. After SNP genotyping and subsequent homozygosity mapping, we identified a single loss-of-function variant by whole-genome sequencing in the canine SLC19A3 gene situated in a 1.7 Mb region of homozygosity on chromosome 25. All ten cases were homozygous carriers of a mutant allele, an indel variant in exon 2, that is predicted to lead to a frameshift and to truncate about 86% of the wild type coding sequence. This study reports a most likely pathogenic variant in SLC19A3 causing a form of SNE in Yorkshire terriers and enables selection against this fatal neurodegenerative recessive disorder. This is the second report of a pathogenic alteration of the SLC19A3 gene in dogs with SNE.

Evaluation of L7-S1 nerve root pathology with low-field MRI in dogs with lumbosacral foraminal stenosis.

OBJECTIVE: To describe low-field MRI findings associated with lumbosacral foraminal stenosis and radiculopathy and correlate these with clinical signs. STUDY DESIGN: Retrospective study. ANIMALS: Client-owned dogs (n = 240) that underwent a clinical examination and standardized MRI protocol of the lumbosacral junction. METHODS: Medical records of dogs with degenerative lumbosacral stenosis with neurological clinical evaluation and MRI of the lumbosacral junction were used to describe imaging pathologies and relate them to clinical status. RESULTS: In total, 480 L7 neuroforamina were evaluated. A loss of foraminal fat signal was identified in 364 of 480 neuroforamina of which 87.9% (n=320) showed also concurrent nerve root changes. Magnetic resonance imaging features of L7 radiculopathy included nerve root enlargement and hyperintensity to surrounding connective tissue in dorsal oblique gradient echo short time inversion recovery sequences and specific changes in shape, size, or position of the nerve root in transverse T1-weighted sequences. Radiculopathy was noted as a consequence of either circumferential (entrapment) or focal (impingement) foraminal stenosis. Lateral vertebral spondylotic and intervertebral facet joint changes were the most common underlying spinal and neuroforaminal pathologies. Clinical signs were present in the ipsilateral hind leg in 85% (n = 65) of dogs with unilateral lumbosacral imaging findings. CONCLUSION: A loss of foraminal fat signal was likely to be associated with L7 radiculopathy and foraminal stenosis. Unilateral lesions were generally associated with clinical signs on the ipsilateral limb. CLINICAL SIGNIFICANCE: Loss of foraminal fat signal revealed by low-field MRI should prompt the assessment of concurrent radiculopathy and underlying stenosis, and in coherence with clinical findings, when is combined with clinical findings, improves the diagnosis of lumbosacral foraminal stenosis.

Leigh-like subacute necrotising encephalopathy in Yorkshire Terriers: neuropathological characterisation, respiratory chain activities and mitochondrial DNA.

Our knowledge of molecular mechanisms underlying mitochondrial disorders in humans has increased considerably during the past two decades. Mitochondrial encephalomyopathies have sporadically been reported in dogs. However, molecular and biochemical data that would lend credence to the suspected mitochondrial origin are largely missing. This study was aimed to characterise a Leigh-like subacute necrotising encephalopathy (SNE) in Yorkshire Terriers and to shed light on its enzymatic and genetic background. The possible resemblance to SNE in Alaskan Huskies and to human Leigh syndrome (LS) was another focus of interest. Eleven terriers with imaging and/or gross evidence of V-shaped, non-contiguous, cyst-like cavitations in the striatum, thalamus and brain stem were included. Neuropathological examinations focussed on muscle, brain pathology and mitochondrial ultrastructure. Further investigations encompassed respiratory-chain activities and the mitochondrial DNA. In contrast to mild non-specific muscle findings, brain pathology featured the stereotypic triad of necrotising grey matter lesions with relative preservation of neurons in the aforementioned regions, multiple cerebral infarcts, and severe patchy Purkinje-cell degeneration in the cerebellar vermis. Two dogs revealed a reduced activity of respiratory-chain-complexes I and IV. Genetic analyses obtained a neutral tRNA-Leu(UUR) A-G-transition only. Neuropathologically, SNE in Yorkshire Terriers is nearly identical to the Alaskan Husky form and very similar to human LS. This study, for the first time, demonstrated that canine SNE can be associated with a combined respiratory chain defect. Mitochondrial tRNA mutations and large genetic rearrangements were excluded as underlying aetiology. Further studies, amongst relevant candidates, should focus on nuclear encoded transcription and translation factors.

Surgical treatment of lumbosacral foraminal stenosis using a lateral approach in twenty dogs with degenerative lumbosacral stenosis.

OBJECTIVES: To describe clinical signs, magnetic resonance imaging (MRI) and surgical findings using a lateral approach to the lumbosacral intervertebral foramen and to evaluate clinical outcomes in dogs with or without concurrent dorsal decompression and annulectomy. STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=20) with degenerative lumbosacral stenosis (DLSS). METHODS: Medical records (2002-2006) of dogs that had lumbosacral lateral foraminotomy alone or in combination with dorsal decompression were reviewed. Degree of dysfunction was assessed separately for each pelvic limb; dogs with unilateral signs were included in group A, those with bilateral signs in group B. Retrieved data were: signalment, history, neurologic status on admission, 3 days, 6 weeks, and 6 months postoperatively, duration of clinical signs, results of MRI, surgical site(s), intraoperative findings, and outcome. RESULTS: Based on the clinical and MRI findings unilateral foraminotomy was performed in 8 dogs, bilateral foraminotomy in 1 dog, unilateral foraminotomy with concurrent dorsal decompression in 7 dogs, and bilateral foraminotomy with concomitant dorsal decompression in 4 dogs. Surgery confirmed the presence of foraminal stenosis in all dogs, with osteophyte formation and soft tissue proliferations being the most common lesions. Outcome was good to excellent in 19 dogs and poor in 1 dog. Mean follow-up was 15.2 months (range, 6-42 months). CONCLUSION: Lateral foraminotomy addresses compressive lesions within exit and middle zones of the lumbosacral foramen. CLINICAL RELEVANCE: Successful surgical management of DLSS is dependent on recognition and correction of each of the compressive lesions within the lumbosacral junction.