Prognosis in meningoencephalitis of unknown origin in dogs: Risk factors associated with survival, clinical relapse, and long-term disability.

BACKGROUND: Meningoencephalitis of unknown origin (MUO) comprises a group of noninfectious inflammatory diseases affecting the central nervous system of dogs. Previous studies have reported individual risk factors for survival but prognostication for MUO remains challenging. OBJECTIVES: Identify clinical prognostic variables in dogs with MUO. ANIMALS: A retrospective study of 447 dogs presented to 2 UK referral hospitals and diagnosed with MUO. METHODS: Medical records of dogs diagnosed with MUO were retrospectively reviewed. Multivariable logistic regression was used for the identification of risk factors for survival and Cox proportional hazards analysis for the identification of risk factors for clinical relapse. RESULTS: Eighty-two percent (366/447) of dogs with presumptive MUO survived to discharge and 63.5% (284/447) were alive at 6 months; 36% of the latter (103/284) had persistent neurological deficits. Breed (pugs; P = .03), epileptic seizures (P < .001), paresis (P < .001), and higher neurodisability scale (NDS) score (P < .001) at presentation were negatively associated with survival to 6 months. Dogs with persistent deficits had higher NDS scores on presentation (P = .001). Median follow-up time was 11 months (interquartile range [IQR], 1-24) and 50.6% (160/316) relapsed during treatment (median time to relapse, 7 months; IQR, 2-15). Incomplete resolution of the clinical signs during the 6 months after diagnosis (P < .001), higher NDS score (P < .001), and longer duration of the clinical signs (P < .001) were associated with relapse. CONCLUSIONS AND CLINICAL IMPORTANCE: Knowledge of risk factors associated with survival, incomplete recovery and clinical relapse in MUO can help guide monitoring and treatment and improve owner communications regarding prognosis for this debilitating disease.

Magnetic resonance imaging prognostic factors for survival and relapse in dogs with meningoencephalitis of unknown origin.

Introduction: Canine meningoencephalitis of unknown origin (MUO) is a debilitating disease associated with high mortality. The prognostic value of magnetic resonance imaging (MRI) findings for predicting survival at 12 months and long-term relapse remains uncertain. Methods: This was a retrospective cohort study evaluating the prognostic value of different MRI variables using multivariable logistic regression and Cox proportional hazards analysis. Results: In total, 138 dogs were presumptively diagnosed with MUO. The most common location for lesions identified on MRI were the white matter tracts of the corona radiata and corpus callosum, followed by the frontal, sensorimotor and temporal cortices. Lower T2 lesion load (p = 0.006, OR = 0.942, CI = 0.902-0.983) was associated with longer survival and higher T1 post-contrast lesion load (p = 0.023, OR = 1.162, CI = 1.021-1.322) was associated with relapse. Discussion: This study has identified prognostic factors that may help identify dogs at higher risk of death and relapse and therefore guide treatment recommendations.

Development of a reliable clinical assessment tool for meningoencephalitis in dogs: The neurodisability scale.

BACKGROUND: Meningoencephalitis of unknown origin (MUO) comprises a group of debilitating inflammatory diseases affecting the central nervous system of dogs. Currently, no validated clinical scale is available for the objective assessment of MUO severity. OBJECTIVES: Design a neurodisability scale (NDS) to grade clinical severity and determine its reliability and whether or not the score at presentation correlates with outcome. ANIMALS: One hundred dogs with MUO were included for retrospective review and 31 dogs were subsequently enrolled for prospective evaluation. METHODS: Medical records were retrospectively reviewed for 100 dogs diagnosed with MUO to identify the most frequent neurological examination findings. The NDS was designed based on these results and evaluated for prospective and retrospective use in a new population of MUO patients (n = 31) by different groups of independent blinded assessors, including calculation of interobserver agreement and association with outcome. RESULTS: The most common clinical signs in MUO patients were used to inform categories for scoring in the NDS: seizure activity, ambulatory status, posture and cerebral, cerebellar, brainstem, and visual functions. The intraclass correlation coefficient (ICC) for prospective use of the NDS was 0.83 (95% confidence interval [CI], 0.68-0.91) indicating good agreement, and moderate agreement was found between prospective and retrospective assessors (ICC, 0.71; 95% CI, 0.56-0.83). No association was found between NDS score and long-term outcome. CONCLUSIONS AND CLINICAL IMPORTANCE: The NDS is a novel clinical measure for objective assessment of neurological dysfunction and showed good reliability when used prospectively in MUO patients but, in this small population, no association with outcome could be identified.

Inflammatory Disease Affecting the Central Nervous System in Dogs: A Retrospective Study in England (2010-2019).

The epidemiology of inflammatory diseases affecting the central nervous system (CNS) in dogs is largely unknown. We aimed to report the relative proportion of different causes of inflammatory disease affecting the CNS in dogs and identify predictors for infectious vs. immune-mediated conditions and predictors for the most common diseases affecting the brain and the spinal cord. This was a retrospective cohort study over a 10-year period in 2 referral institutions using multivariable and multinomial logistic regression for identification of risk factors. In total, 1,140 client-owned dogs diagnosed with inflammatory disease affecting the CNS were included. Fifteen different diagnoses were identified, with immune-mediated (83.6%) disease being more common than infectious conditions (16.4%). The most common immune-mediated conditions diagnosed were meningoencephalitis of unknown origin (47.5%) and steroid-responsive meningitis-arteritis (30.7%), and the most common infectious conditions were discospondylitis (9.3%) and otogenic intracranial infection (2.2%). Older age (p < 0.001, OR = 1.019, 95% CI: 1.014-1.024), higher body weight (p < 0.001, OR = 1.049, 95% CI: 1.025-1.074), male sex (p = 0.009, OR = 1.685, 95% CI: 1.141-2.488), longer duration of the clinical signs before presentation (p < 0.001, OR = 1.011, 95% CI: 1.006-1.017), progressive nature of the clinical signs (p < 0.001, OR = 2.295, 95% CI: 1.463-3.599), identification of a possibly associated preceding event (p = 0.0012, OR = 1.93, 95% CI: 1.159-3.213), and hyperesthesia on presentation (p < 0.001, OR = 2.303, 95% CI: 1.528-3.473) were associated with a diagnosis of infectious diseases. Our data shows that immune-mediated diseases are more common than infectious conditions as a cause for inflammatory CNS disease in dogs. The risk factors for the most common diagnoses were identified from signalment, history, and findings of the physical and neurological examinations to give valuable information that can guide clinicians with their investigations.

Identifying practical approaches to the normalisation of interprofessional collaboration in rural hospitals: A qualitative study among health professionals.

This qualitative study explores the ideas and experiences of interprofessional collaboration (IPC) among health professionals in rural public hospitals and to propagate its normalization into practice by identifying existing or suggested solutions. The literature focuses largely on the barriers and facilitators to IPC in metropolitan areas and there is room to identify more practical responses for implementing solutions. Semi-structured interviews were conducted with 13 healthcare professionals (October 2018-March 2019). Interviews were audio-recorded, transcribed and underwent thematic analysis to identify themes derived from the dataset. Using the lens of the Normalization Process Theory (NPT) allowed for amalgamation of participant ideas and identification of solutions to implement IPC in practice. Participants' definitions of IPC and Interprofessional Teamwork were incongruous with the current literature, however when provided with formal definitions, participants agreed that they both participated and observed IPC with varying degrees of success. Factors influencing this success included good working relationships and positive workplace cultures, having an understanding of each professions' roles and needs and the hierarchy of professions in conjunction with attitudes of senior healthcare professionals. Solutions to improved IPC and its normalization included induction processes and informal introductions, formalized interprofessional interactions, interprofessional education and positive leadership, such as the 'assertive followership model'. Analyzed in the framework of the normalization process theory, this research shows that IPC is increasingly becoming a coherent, integrated aspect of the healthcare system but there is room for improvement, and cognitive participation in IPC varies across healthcare professionsals. In order to facilitate the normalization process, program and policy makers, hospital administrations and professional associations could consider formalized interprofessional team interactions, formalizing IPC through simple introductions, interprofessional education and positive leadership. Future research could explore through the NPT specific areas of care that benefit from IPC implementation such as community aged-care.

CT-guided drainage of a brainstem abscess in a cat as an emergency treatment procedure.

CASE SUMMARY: A 3-year-old male neutered domestic shorthair cat was presented with a 1-week progressive and rapidly deteriorating history of lethargy and abnormal behaviour. Neurolocalisation indicated multifocal intracranial lesions (right oculomotor nerve, brainstem [obtundation, non-ambulatory tetraparesis, vestibular dysfunction and intermittent decerebrate rigidity] and possibly the thalamus [left-sided pleurothotonus]), or more likely a single brainstem lesion with mass effect. MRI of the brain demonstrated a brainstem abscess causing severe dorsal displacement particularly affecting the pons and the medulla oblongata causing cerebellar vermis herniation through the foramen magnum. CT-guided free-hand technique drainage of the brain abscess was performed and broad spectrum antibiotics were started based on sensitivity results. The cat recovered uneventfully from anaesthesia displaying marked improvement immediately after the procedure. Antibiotics were continued for 8 months; repeat imaging prior to withdrawal found complete resolution of the brainstem abscess. RELEVANCE AND NOVEL INFORMATION: Free-hand CT-guided drainage of a brainstem abscess is not without risk; however, in this case it led to significant clinical improvement and stabilisation likely owing to reduced intracranial pressure. It also provided a diagnostic sample that allowed successful medical treatment planning and outcome. To our knowledge, this is the first report describing the successful management of a brainstem abscess by CT-guided drainage in the veterinary literature. It suggests that stereotactic drainage followed by medical therapy can be considered a successful therapeutic alternative to brain surgery or medical treatment alone, providing an emergency treatment in cases of acute brainstem dysfunction.

Evaluation of concurrent analysis of cerebrospinal fluid samples collected from the cerebellomedullary cistern and lumbar subarachnoid space for the diagnosis of steroid-responsive meningitis arteritis in dogs.

OBJECTIVE: To evaluate whether concurrent analysis of CSF samples from 2 collection sites (cerebellomedullary cistern [CMC] and lumbar subarachnoid space [LSS]) versus only 1 site could improve the diagnostic sensitivity of CSF analysis for dogs with suspected steroid-responsive meningitis arteritis (SRMA). ANIMALS: 111 client-owned dogs with SRMA diagnosed at 3 veterinary referral hospitals between 2011 and 2017. PROCEDURES: Only dogs with CSF collected from both sites (CMC and LSS) and with no previous history of corticosteroid administration were included. Medical record data and logistic regression were used to identify factors associated with having a total nucleated cell concentration (TNCC) within the reference interval in a CSF sample from 1 collection site. RESULTS: The TNCC was within the reference interval (TNCC < 5 cells/µL) in the CSF sample from 1 collection site for 8 of 111 (7%) dogs and was only slightly high (TNCC, 5 to 9 cells/µL) in the sample from 1 or both sites for 10 (11%) other dogs. For each of these 18 dogs, results for samples from 1 site were consistent with SRMA. The proportion of CSF samples that had a TNCC within the reference interval was comparable between sites. As age increased, so did the risk of having an unremarkable TNCC in the CSF sample from 1 site, albeit only slightly (OR, 1.08; 95% confidence interval, 1.01 to 1.16). CONCLUSIONS AND CLINICAL RELEVANCE: CSF samples from both the CMC and LSS should be analyzed when evaluating dogs with suspected SRMA to improve the chance of detecting a high TNCC.

Progressive Structural Defects in Canine Centronuclear Myopathy Indicate a Role for HACD1 in Maintaining Skeletal Muscle Membrane Systems.

Mutations in HACD1/PTPLA cause recessive congenital myopathies in humans and dogs. Hydroxyacyl-coA dehydratases are required for elongation of very long chain fatty acids, and HACD1 has a role in early myogenesis, but the functions of this striated muscle-specific enzyme in more differentiated skeletal muscle remain unknown. Canine HACD1 deficiency is histopathologically classified as a centronuclear myopathy (CNM). We investigated the hypothesis that muscle from HACD1-deficient dogs has membrane abnormalities in common with CNMs with different genetic causes. We found progressive changes in tubuloreticular and sarcolemmal membranes and mislocalized triads and mitochondria in skeletal muscle from animals deficient in HACD1. Furthermore, comparable membranous abnormalities in cultured HACD1-deficient myotubes provide additional evidence that these defects are a primary consequence of altered HACD1 expression. Our novel findings, including T-tubule dilatation and disorganization, associated with defects in this additional CNM-associated gene provide a definitive pathophysiologic link with these disorders, confirm that dogs deficient in HACD1 are relevant models, and strengthen the evidence for a unifying pathogenesis in CNMs via defective membrane trafficking and excitation-contraction coupling in muscle. These results build on previous work by determining further functional roles of HACD1 in muscle and provide new insight into the pathology and pathogenetic mechanisms of HACD1 CNM. Consequently, alterations in membrane properties associated with HACD1 mutations should be investigated in humans with related phenotypes.

HACD1, a regulator of membrane composition and fluidity, promotes myoblast fusion and skeletal muscle growth.

The reduced diameter of skeletal myofibres is a hallmark of several congenital myopathies, yet the underlying cellular and molecular mechanisms remain elusive. In this study, we investigate the role of HACD1/PTPLA, which is involved in the elongation of the very long chain fatty acids, in muscle fibre formation. In humans and dogs, HACD1 deficiency leads to a congenital myopathy with fibre size disproportion associated with a generalized muscle weakness. Through analysis of HACD1-deficient Labradors, Hacd1-knockout mice, and Hacd1-deficient myoblasts, we provide evidence that HACD1 promotes myoblast fusion during muscle development and regeneration. We further demonstrate that in normal differentiating myoblasts, expression of the catalytically active HACD1 isoform, which is encoded by a muscle-enriched splice variant, yields decreased lysophosphatidylcholine content, a potent inhibitor of myoblast fusion, and increased concentrations of ≥ C18 and monounsaturated fatty acids of phospholipids. These lipid modifications correlate with a reduction in plasma membrane rigidity. In conclusion, we propose that fusion impairment constitutes a novel, non-exclusive pathological mechanism operating in congenital myopathies and reveal that HACD1 is a key regulator of a lipid-dependent muscle fibre growth mechanism.

Imaging diagnosis--spinal cord hemangioma in two dogs.

Intramedullary masses are a dilemma due to the limited access for a nonsurgical biopsy, thus, accurate imaging characterization is crucial. Magnetic resonance imaging findings of two confirmed canine thoracic intramedullary hemangiomas are described. A capillary hemangioma was of mixed intensity but predominantly T2-hyperintense and mildly T1-hyperintense to spinal cord with strong contrast enhancement. A cavernous hemangioma had a target-like appearance in both T1-weighted (T1w) and T2-weighted (T2w) images. In T2w images there was a small isointense center surrounded by a relatively large hyperintense area. In T1w images, there was a large isointense centre with a relatively small hyperintense periphery. Such characteristics should prioritize hemangioma as a consideration in a progressive myelopathy due to an intramedullary mass.

A duchenne muscular dystrophy gene hot spot mutation in dystrophin-deficient cavalier king charles spaniels is amenable to exon 51 skipping.

BACKGROUND: Duchenne muscular dystrophy (DMD), which afflicts 1 in 3500 boys, is one of the most common genetic disorders of children. This fatal degenerative condition is caused by an absence or deficiency of dystrophin in striated muscle. Most affected patients have inherited or spontaneous deletions in the dystrophin gene that disrupt the reading frame resulting in unstable truncated products. For these patients, restoration of the reading frame via antisense oligonucleotide-mediated exon skipping is a promising therapeutic approach. The major DMD deletion "hot spot" is found between exons 45 and 53, and skipping exon 51 in particular is predicted to ameliorate the dystrophic phenotype in the greatest number of patients. Currently the mdx mouse is the most widely used animal model of DMD, although its mild phenotype limits its suitability in clinical trials. The Golden Retriever muscular dystrophy (GRMD) model has a severe phenotype, but due to its large size, is expensive to use. Both these models have mutations in regions of the dystrophin gene distant from the commonly mutated DMD "hot spot". METHODOLOGY/PRINCIPAL FINDINGS: Here we describe the severe phenotype, histopathological findings, and molecular analysis of Cavalier King Charles Spaniels with dystrophin-deficient muscular dystrophy (CKCS-MD). The dogs harbour a missense mutation in the 5' donor splice site of exon 50 that results in deletion of exon 50 in mRNA transcripts and a predicted premature truncation of the translated protein. Antisense oligonucleotide-mediated skipping of exon 51 in cultured myoblasts from an affected dog restored the reading frame and protein expression. CONCLUSIONS/SIGNIFICANCE: Given the small size of the breed, the amiable temperament and the nature of the mutation, we propose that CKCS-MD is a valuable new model for clinical trials of antisense oligonucleotide-induced exon skipping and other therapeutic approaches for DMD.

Molecular and electrophysiological characterization of transient receptor potential ion channels in the primary murine megakaryocyte.

The molecular identity of platelet Ca(2+) entry pathways is controversial. Furthermore, the extent to which Ca(2+)-permeable ion channels are functional in these tiny, anucleate cells is difficult to assess by direct electrophysiological measurements. Recent work has highlighted how the primary megakaryocyte represents a bona fide surrogate for studies of platelet signalling, including patch clamp recordings of ionic conductances. We have now screened for all known members of the transient receptor potential (TRP) family of non-selective cation channels in murine megakaryocytes following individual selection of these rare marrow cells using glass micropipettes. RT-PCR detected messages for TRPC6 and TRPC1, which have been reported in platelets and megakaryocytic cell lines, and TRPM1, TRPM2 and TRPM7, which to date have not been demonstrated in cells of megakaryocytic/platelet lineage. Electrophysiological recordings demonstrated the presence of functional TRPM7, a constitutively active cation channel sensitive to intracellular Mg(2+), and TRPM2, an ADP-ribose-dependent cation channel activated by oxidative stress. In addition, the electrophysiological and pharmacological properties of the non-selective cation channels stimulated by the physiological agonist ADP are consistent with a major role for TRPC6 in this G-protein-coupled receptor-dependent Ca(2+) influx pathway. This study defines for the first time the principal TRP channels within the primary megakaryocyte, which represent candidates for Ca(2+) influx pathways activated by a diverse range of stimuli in the platelet and megakaryocyte.