CD8+ T cells specific for an immunodominant SARS-CoV-2 nucleocapsid epitope display high naive precursor frequency and TCR promiscuity.

To better understand primary and recall T cell responses during coronavirus disease 2019 (COVID-19), it is important to examine unmanipulated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T cells. By using peptide-human leukocyte antigen (HLA) tetramers for direct ex vivo analysis, we characterized CD8+ T cells specific for SARS-CoV-2 epitopes in COVID-19 patients and unexposed individuals. Unlike CD8+ T cells directed toward subdominant epitopes (B7/N257, A2/S269, and A24/S1,208) CD8+ T cells specific for the immunodominant B7/N105 epitope were detected at high frequencies in pre-pandemic samples and at increased frequencies during acute COVID-19 and convalescence. SARS-CoV-2-specific CD8+ T cells in pre-pandemic samples from children, adults, and elderly individuals predominantly displayed a naive phenotype, indicating a lack of previous cross-reactive exposures. T cell receptor (TCR) analyses revealed diverse TCRαß repertoires and promiscuous αß-TCR pairing within B7/N105+CD8+ T cells. Our study demonstrates high naive precursor frequency and TCRαß diversity within immunodominant B7/N105-specific CD8+ T cells and provides insight into SARS-CoV-2-specific T cell origins and subsequent responses.

Towards a point-of-care multimodal spectroscopy instrument for the evaluation of human cardiac tissue.

To demonstrate that point-of-care multimodal spectroscopy using Near-Infrared (NIR) and Raman Spectroscopy (RS) can be used to diagnose human heart tissue. We generated 105 spectroscopic scans, which comprised 4 NIR and 3 RS scans per sample to generate a "multimodal spectroscopic scan" (MSS) for each heart, done across 15 patients, 5 each from the dilated cardiomyopathy (DCM), Ischaemic Heart Disease (IHD) and Normal pathologies. Each of the MSS scans was undertaken in 3 s. Data were entered into machine learning (ML) algorithms to assess accuracy of MSS in diagnosing tissue type. The median age was 50 years (IQR 49-52) for IHD, 47 (IQR 45-50) for DCM and 36 (IQR 33-52) for healthy patients (p = 0.35), 60% of which were male. MSS identified key differences in IHD, DCM and normal heart samples in regions typically associated with fibrosis and collagen (NIR wavenumbers: 1433, 1509, 1581, 1689 and 1725 nm; RS wavelengths: 1658, 1450 and 1330 cm-1). In principal component (PC) analyses, these differences explained 99.2% of the variation in 4 PCs for NIR, 81.6% in 10 PCs for Raman, and 99.0% in 26 PCs for multimodal spectroscopic signatures. Using a stack machine learning algorithm with combined NIR and Raman data, our model had a precision of 96.9%, recall of 96.6%, specificity of 98.2% and Area Under Curve (AUC) of 0.989 (Table 1). NIR and Raman modalities alone had similar levels of precision at 94.4% and 89.8% respectively (Table 1). MSS combined with ML showed accuracy of 90% for detecting dilated cardiomyopathy, 100% for ischaemic heart disease and 100% for diagnosing healthy tissue. Multimodal spectroscopic signatures, based on NIR and Raman spectroscopy, could provide cardiac tissue scans in 3-s to aid accurate diagnoses of fibrosis in IHD, DCM and normal hearts. Table 1 Machine learning performance metrics for validation data sets of (a) Near-Infrared (NIR), (b) Raman and (c and d) multimodal data using logistic regression (LR), stochastic gradient descent (SGD) and support vector machines (SVM), with combined "stack" (LR + SGD + SVM) AUC Precision Recall Specificity (a) NIR model  Logistic regression 0.980 0.944 0.933 0.967  SGD 0.550 0.281 0.400 0.700  SVM 0.840 0.806 0.800 0.900  Stack 0.933 0.794 0.800 0.900 (b) Raman model  Logistic regression 0.985 0.940 0.929 0.960  SGD 0.892 0.869 0.857 0.932  SVM 0.992 0.940 0.929 0.960  Stack 0.954 0.869 0.857 0.932 (c) MSS: multimodal (NIR + Raman) to detect DCM vs. IHD vs. normal patients  Logistic regression 0.975 0.841 0.828 0.917  SGD 0.847 0.803 0.793 0.899  SVM 0.971 0.853 0.828 0.917  Stack 0.961 0.853 0.828 0.917 (d) MSS: multimodal (NIR + Raman) to detect pathological vs. normal patients  Logistic regression 0.961 0.969 0.966 0.984  SGD 0.944 0.967 0.966 0.923  SVM 1.000 1.000 1.000 1.000  Stack 1.000 0.944 0.931 0.969 Bold values indicate values obtained from the stack algorithm and used for analyses.

Outcomes After Liver Transplantation With Incidental Cholangiocarcinoma.

Cholangiocarcinoma (CCA) is currently a contraindication to liver transplantation (LT) in the United Kingdom (UK). Incidental CCA occurs rarely in some patients undergoing LT. We report on retrospective outcomes of patients with incidental CCA from six UK LT centres. Cases were identified from pathology records. Data regarding tumour characteristics and post-transplant survival were collected. CCA was classified by TNM staging and anatomical location. 95 patients who underwent LT between 1988-2020 were identified. Median follow-up after LT was 2.1 years (14 days-18.6 years). Most patients were male (68.4%), median age at LT was 53 (IQR 46-62), and the majority had underlying PSC (61%). Overall median survival after LT was 4.4 years. Survival differed by tumour site: 1-, 3-, and 5-year estimated survival was 82.1%, 68.7%, and 57.1%, respectively, in intrahepatic CCA (n = 40) and 58.5%, 42.6%, and 30.2% in perihilar CCA (n = 42; p = 0.06). 1-, 3-, and 5-year estimated survival was 95.8%, 86.5%, and 80.6%, respectively, in pT1 tumours (28.2% of cohort), and 65.8%, 44.7%, and 31.1%, respectively, in pT2-4 (p = 0.018). Survival after LT for recipients with incidental CCA is inferior compared to usual outcomes for LT in the United Kingdom. LT for earlier stage CCA has similar survival to LT for hepatocellular cancer, and intrahepatic CCAs have better survival compared to perihilar CCAs. These observations may support LT for CCA in selected cases.

Generalized myoclonic epilepsy with photosensitivity in juvenile dogs caused by a defective DIRAS family GTPase 1.

The clinical and electroencephalographic features of a canine generalized myoclonic epilepsy with photosensitivity and onset in young Rhodesian Ridgeback dogs (6 wk to 18 mo) are described. A fully penetrant recessive 4-bp deletion was identified in the DIRAS family GTPase 1 (DIRAS1) gene with an altered expression pattern of DIRAS1 protein in the affected brain. This neuronal DIRAS1 gene with a proposed role in cholinergic transmission provides not only a candidate for human myoclonic epilepsy but also insights into the disease etiology, while establishing a spontaneous model for future intervention studies and functional characterization.

Electroencephalographic, physiologic and behavioural responses during cervical dislocation euthanasia in turkeys.

BACKGROUND: There is a critical need to develop appropriate on-farm euthanasia methods for poultry species. Euthanasia methods should affect the brain first causing insensibility, followed by cardiorespiratory arrest. Neck or cervical dislocation methods, either manual (CD) or mechanical (MCD), are reported to cause a prolonged time to loss of sensibility and death with inconsistent results upon application, especially MCD methods. However, there is limited information on cervical dislocation in turkeys. The overall objective of this study was to assess the welfare implications of CD and a newly developed MCD device for euthanasia of cull turkeys in comparison with intravenous (IV) pentobarbital sodium (1 mL/4.5 kg), the gold standard euthanasia method. Time to death using electroencephalographic (EEG) and behavioural responses were monitored in eight and eighteen week-old turkeys for five minutes after each euthanasia method application. Spectral analyses of EEG responses and onset of isoelectric EEGs were compared to baseline EEG recordings of birds under anesthesia and behavioural responses were studied among euthanasia treatments. A significant decrease in brain activity frequencies analysis and isoelectric EEG were recorded as time of brain death. RESULTS: All turkeys euthanized with IV pentobarbital sodium presented a rapid and irreversible decrease in the EEG activity at approximately 30s post-injection with minimal behavioural responses. CD and MCD methods caused EEG responses consistent with brain death at approximately 120 s and 300 s, respectively. Additionally, isoelectric EEGs resulted in all pentobarbital sodium and CD groups, but only in 54 and 88% of the eight and eighteen week-old turkeys in the MCD groups, respectively. There were few clear patterns of behavioural responses after CD and MCD application. However, cessation of body movement and time to isoelectric EEG after CD application were positively correlated. CONCLUSIONS: Use of CD and MCD resulted in a prolonged time to death in both age groups of turkeys. MCD application presents a number of welfare risks based on electroencephalographic and behavioural findings. Intravenous pentobarbital sodium induced rapid brain death, but possesses several on-farm limitations. To develop improvements in cervical dislocation methods, further investigations into combined or alternative methods are required to reduce the prolonged time to insensibility and death.

Diagnostic imaging characteristics of canine infectious sacroiliitis.

Infectious sacroiliitis has not been described in dogs. This retrospective case series describes the presentation, diagnostic imaging characteristics, and outcomes of 2 canine patients with infectious sacroiliitis. Selection criteria included presentation with back pain from 2010 to 2017, diagnostic imaging of the sacroiliac joints, and short- and long-term response to antibiotic therapy. Medical records, and magnetic resonance imaging (MRI) were reviewed by a Board-certified veterinary radiologist, a neurologist, and a small animal intern. Two dogs met the inclusion criteria. Magnetic resonance imaging revealed unilaterally wide and irregular sacroiliac joint spaces, with juxta-articular soft tissue contrast enhancement and bone marrow edema. One patient had a communicating abscess of the psoas muscle, which cultured positive for Pasteurella canis. Following treatment with pain relief medications and antibiotics, both patients made a complete clinical recovery, with no signs of lameness 2 to 4 weeks after cessation of treatment, and no lameness reported by the owner afterwards. Infectious sacroiliitis should be considered when dogs are presented with lumbosacral pain.

Caractéristiques de l'imagerie diagnostique de la sacro-iliite infectieuse canine. La sacro-iliite infectieuse n'a pas été décrite chez les chiens. Cette série de cas rétrospectifs décrit la présentation, les caractéristiques de l'imagerie diagnostique et les résultats de deux patients canins atteints de sacro-iliite infectieuse. Les critères de sélection incluaient la présentation avec des douleurs au dos de 2010 à 2017, l'imagerie diagnostique des articulations sacro-iliaques et une réponse à court et à long terme à la thérapie aux antibiotiques. Les dossiers médicaux et l'imagerie à résonance magnétique (IRM) ont été évalués par un radiologiste vétérinaire agréé, un neurologue et un interne en médecine des petits animaux. Deux chiens satisfaisaient aux critères d'inclusion. L'imagerie à résonance magnétique a révélé des espaces d'articulation sacro-iliaque unilatéralement larges et irréguliers avec une augmentation de contraste des tissus mous juxta-articulaires et de l'oedème de la moelle osseuse. Le patient 2 avait un abcès communiquant du muscle psoas qui a révélé la présence de Pasteurella canis à la culture. Après un traitement avec des médicaments analgésiques et des antibiotiques, les deux patients ont connu un rétablissement clinique complet sans signes de boiterie de 2 à 4 semaines après l'arrêt du traitement et aucune boiterie n'a été signalée par la suite par le propriétaire. La sacro-iliite infectieuse devrait être considérée lorsque les chiens sont présentés avec des douleurs lombo-sacrées.(Traduit par Isabelle Vallières).

Magnetic resonance imaging muscle lesions in presumptive canine fibrocartilaginous embolic myelopathy.

This retrospective cohort study reports the observation of magnetic resonance imaging (MRI) epaxial muscle hyperintensity in dogs diagnosed with presumptive fibrocartilaginous embolic myelopathy (FCEM) (n = 61). It further reports the observation of vertebral column hyperesthesia lasting > 12 hours. The hypothesis tested was that the finding of MRI epaxial muscle hyperintensity correlated with dogs presenting with hyperesthesia. Client-owned dogs diagnosed with presumptive FCEM by specific MRI criteria were included. Statistical analysis was performed using Fisher's exact test. Twenty-three percent (14/61) of MRIs displayed abnormal muscle hyperintensity and 43% (26/61) exhibited vertebral column hyperesthesia. No relationship was found between muscle hyperintensity and pain persisting beyond 12 hours. The muscle hyperintensity remains of unknown significance. That 43% of presumptive FCEM cases have prolonged signs of pain is a higher prevalence than previously reported, and may affect clinical differential diagnoses. This is especially significant in cases in which MRI is not possible and a presumptive diagnosis must be based on the clinical signs.

Imagerie par résonance magnétique des lésions des muscles dans la myélopathie embolique fibrocartilagineuse canine présumée. Cette étude rétrospective de cohorte signale les observations de l'imagerie par résonance magnétique (IRM) pour l'hyperintensité du muscle épaxial chez les chiens diagnostiqués avec une myélopathie embolique fibrocartilagineuse (MEFC) présumée (n = 61). Elle signale aussi l'observation de l'hyperesthésie de la colonne vertébrale durant > 12 heures. L'hypothèse qui a été testée était qu'il y avait une corrélation entre l'observation de l'hyperintensité du muscle épaxial par IRM et les chiens présentés avec de l'hyperesthésie. Les chiens appartenant à des clients pour lesquels un diagnostic présomptif de MEFC avait été posé à l'aide du critère spécifique de l'IRM ont été inclus. L'analyse statistique a été réalisée en utilisant le test exact de Fisher. Vingt-trois pour cent (14/61) des IRM affichaient une hyperintensité anormale du muscle et 43 % (26/61) présentaient de l'hypersthésie de la colonne vertébrale. Aucun lien n'a été trouvé entre l'hyperintensité musculaire et la douleur persistant au-delà de 12 heures. La signification de l'hyperintensité musculaire est toujours inconnue. Le taux de 43 % de cas présomptifs de MEFC affichant des signes de douleur prolongée représente une prévalence supérieure aux données déjà signalées et pourrait affecter les diagnostics cliniques différentiels. Ce fait revêt une importance particulière lorsque l'IRM n'est pas possible et qu'un diagnostic présomptif doit se baser sur les signes cliniques.(Traduit par Isabelle Vallières).

Association between tumour infiltrating lymphocytes, histotype and clinical outcome in epithelial ovarian cancer.

BACKGROUND: There is evidence that some ovarian tumours evoke an immune response, which can be assessed by tumour infiltrating lymphocytes (TILs). To facilitate adoption of TILs as a clinical biomarker, a standardised method for their H&E visual evaluation has been validated in breast cancer. METHODS: We sought to investigate the prognostic significance of TILs in a study of 953 invasive epithelial ovarian cancer tumour samples, both primary and metastatic, from 707 patients from the prospective population-based SEARCH study. TILs were analysed using a standardised method based on H&E staining producing a percentage score for stromal and intratumoral compartments. We used Cox regression to estimate hazard ratios of the association between TILs and survival. RESULTS: The extent of stromal and intra-tumoral TILs were correlated in the primary tumours (n = 679, Spearman's rank correlation = 0.60, P < 0.001) with a similar correlation in secondary tumours (n = 224, Spearman's rank correlation = 0.62, P < 0.001). There was a weak correlation between stromal TIL levels in primary and secondary tumour samples (Spearman's rank correlation = 0.29, P < 0.001) and intra-tumoral TIL levels in primary and secondary tumour samples (Spearman's rank correlation = 0.19, P = 0.0094). The extent of stromal TILs differed between histotypes (Pearson chi2 (12d.f.) 54.1, P < 0.0001) with higher levels of stromal infiltration in the high-grade serous and endometriod cases. A significant association was observed for higher intratumoral TIL levels and a favourable prognosis (HR 0.74 95% CI 0.55-1.00 p = 0.047). CONCLUSION: This study is the largest collection of epithelial ovarian tumour samples evaluated for TILs. We have shown that stromal and intratumoral TIL levels are correlated and that their levels correlate with clinical variables such as tumour histological subtype. We have also shown that increased levels of both intratumoral and stromal TILs are associated with a better prognosis; however, this is only statistically significant for intratumoral TILs. This study suggests that a clinically useful immune prognostic indicator in epithelial ovarian cancer could be developed using this technique.

Canine atlantoaxial optimal safe implantation corridors - description and validation of a novel 3D presurgical planning method using OsiriX™.

BACKGROUND: Canine ventral atlantoaxial (AA) stabilization is most commonly performed in very small dogs and is technically challenging due to extremely narrow bone corridors. Multiple implantation sites have been suggested but detailed anatomical studies investigating these sites are lacking and therefore current surgical guidelines are based upon approximate anatomical landmarks. In order to study AA optimal safe implantation corridors (OSICs), we developed a method based on computed tomography (CT) and semi-automated three-dimensional (3D) mathematical modelling using OsiriX™ and Microsoft®Excel software. The objectives of this study were 1- to provide a detailed description of the bone corridor analysis method and 2- to assess the reproducibility of the method. CT images of the craniocervical junction were prospectively obtained in 27 dogs and our method of OSIC analysis was applied in all dogs. For each dog, 13 optimal implant sites were simulated via geometrical simplification of the bone corridors. Each implant 3D position was then defined with respect to anatomical axes using 2 projected angles (ProjA). The safety margins around each implant were also estimated with angles (SafA) measured in 4 orthogonal directions. A sample of 12 simulated implants was randomly selected and each mathematically calculated angle was compared to direct measurements obtained within OsiriX™ from 2 observers repeated twice. The landmarks simulating anatomical axes were also positioned 4 times to determine their effect on ProjA reproducibility. RESULTS: OsiriX could be used successfully to simulate optimal implant positions in all cases. There was excellent agreement between the calculated and measured values for both ProjA (ρc = 0.9986) and SafA (ρc = 0.9996). Absolute differences between calculated and measured values were respectively [ProjA = 0.44 ± 0.53°; SafA = 0.27 ± 0.25°] and [ProjA = 0.26 ± 0.21°; SafA = 0.18 ± 0.18°] for each observer. The 95 % tolerance interval comparing ProjA obtained with 4 different sets of anatomical axis landmarks was [-1.62°, 1.61°] which was considered appropriate for clinical use. CONCLUSIONS: A new method for determination of optimal implant placement is provided. Semi-automated calculation of optimal implant 3D positions could be further developed to facilitate preoperative planning and to generate large descriptive anatomical datasets.

Oral challenge vs routine care to assess low-risk penicillin allergy in critically ill hospital patients (ORACLE): a pilot safety and feasibility randomised controlled trial.

PURPOSE: Critically ill patients are vulnerable to penicillin allergy labels that may be incorrect. The validity of skin testing in intensive care units (ICUs) is uncertain. Many penicillin allergy labels are low risk, and validated tools exist to identify those amenable to direct oral challenge. This pilot randomised controlled trial explored the feasibility, safety, and validity of direct enteral challenge for low-risk penicillin allergy labels in critical illness. METHODS: Consenting patients with a low-risk penicillin allergy label (PAL) (PEN-FAST risk assessment score < 3) in four ICUs (Melbourne, Australia) were randomised 1:1 to penicillin (250 mg amoxicillin or implicated penicillin) direct enteral challenge versus routine care (2-h post-randomisation observation for each arm). Repeat challenge was performed post -ICU in the intervention arm. Patients were reviewed at 24 h and 5 days after each challenge/observation. RESULTS: We screened 533 patients. 130 (24.4%) were eligible and 80/130 (61.5%) enrolled (age median 64.5 years (interquartile range, IQR 53.5, 74), PEN-FAST median 1 (IQR 0,1)), with 40 (50%) randomised to direct enteral challenge. A positive challenge rate of 2.5% was identified. No antibiotic-associated serious adverse events were identified. 32/40 (80%) received a repeat challenge (zero positive). Post-randomisation, 13 (32%) of the intervention arm and 4 (10%) of the control arm received penicillin (odds ratio, OR 4.33 [1.27, 14.78] p = 0.019). CONCLUSION: These findings support the safety, validity, and feasibility of direct enteral challenge for critically ill patients with PEN-FAST assessed low-risk penicillin allergy. The absence of false negative results was confirmed by subsequent negative repeat challenges. A relatively low recruitment to screened ratio suggests that more inclusive eligibility criteria and integration of allergy assessment into routine ICU processes are needed to optimise allergy delabelling in critical illness.

Immunotherapy in the context of immune-specialized environment of brain metastases.

Brain metastases (BrM) develop in 20-40% of patients with advanced cancer. They mainly originate from lung cancer, melanoma, breast cancer, and renal cell carcinoma, and are associated with a poor prognosis. While patients with BrM traditionally lack effective treatment options, immunotherapy is increasingly gaining in importance in this group of patients, with clinical trials in the past decade demonstrating the efficacy and safety of immune checkpoint blockade in BrM originating from specific tumor types, foremost melanoma. The brain is an immune-specialized environment with several unique molecular, cellular, and anatomical features that affect immune responses, including those against tumors. In this review we discuss the potential role that some of these unique characteristics may play in the efficacy of immunotherapy, mainly focusing on the lymphatic drainage in the brain and the role of systemic anti-tumor immunity that develops due to the presence of concurrent extracranial disease in addition to BrM.

Tools and techniques for classifying behaviours in canine epilepsy.

Idiopathic epilepsy is the most common neurological disease in dogs. Similar to humans, dogs with epilepsy often experience behavioural comorbidities such as increased fear, anxiety, and aggression, as reported by their caregivers. Investigations of behaviour in canine epilepsy have yet to untangle interictal and pre and postictal behaviours, prodromal changes, and seizure-precipitating factors. Under-recognition of absence and focal seizures further complicates these assessments. These complex behavioural presentations in combination with caring for an epileptic animal have a significant negative impact on the dog's and caregiver's quality of life. Despite the growing recognition of behavioural comorbidities and their impact on quality of life in dogs with epilepsy, few objective research methods for classifying and quantifying canine behaviour exist. This narrative review examines the strengths, limitations, and granularity of three tools used in the investigation of canine behaviour and epilepsy; questionnaires, electroencephalography, and actigraphy. It suggests that a prospective combination of these three tools has the potential to offer improvements to the objective classification and quantification of canine behaviour in epilepsy.

Feasibility of in-home electroencephalographic and actigraphy recordings in dogs.

Introduction: Idiopathic epilepsy is a prevalent neurological disease in dogs. Dogs with epilepsy often present with behavioral comorbidities such as aggression, anxiety, and fear. These behaviors are consistent with pre, post, or interictal behaviors, prodromal changes, seizure-precipitating factors, or absence and focal seizures. The overlap in behavior presentations and lack of objective research methods for quantifying and classifying canine behavior makes determining the cause difficult. Behavioral comorbidities in addition to the task of caring for an epileptic animal have a significant negative impact on dog and caregiver quality of life. Methods: This pilot study aimed to assess the feasibility of a novel technology combination for behavior classification and epileptic seizure detection for a minimum 24-h recording in the dog's home environment. It was expected that combining electroencephalography (EEG), actigraphy, and questionnaires would be feasible in the majority of trials. A convenience sample of 10 community-owned dogs was instrumented with wireless video-EEG and actigraphy for up to 48 h of recording at their caregiver's home. Three questionnaires (maximum 137 questions) were completed over the recording period by caregivers to describe their dog's everyday behavior and habits. Results: Six of the 10 included dogs had combined EEG and actigraphy recordings for a minimum of 24 h. Discussion: This shows that in-home EEG and actigraphy recordings are possible in community-owned dogs and provides a basis for a prospective study examining the same technology combination in a larger sample size.

Biglycan and reduced glycolysis are associated with breast cancer cell dormancy in the brain.

Exit of quiescent disseminated cancer cells from dormancy is thought to be responsible for metastatic relapse and a better understanding of dormancy could pave the way for novel therapeutic approaches. We used an in vivo model of triple negative breast cancer brain metastasis to identify differences in transcriptional profiles between dormant and proliferating cancer cells in the brain. BGN gene, encoding a small proteoglycan biglycan, was strongly upregulated in dormant cancer cells in vivo. BGN expression was significantly downregulated in patient brain metastases as compared to the matched primary breast tumors and BGN overexpression in cancer cells inhibited their growth in vitro and in vivo. Dormant cancer cells were further characterized by a reduced expression of glycolysis genes in vivo, and inhibition of glycolysis in vitro resulted in a reversible growth arrest reminiscent of dormancy. Our study identified mechanisms that could be targeted to induce/maintain cancer dormancy and thereby prevent metastatic relapse.

Survey of electroencephalography usage and techniques for dogs.

Background: Canine epilepsy is a chronic common neurologic condition where seizures may be underreported. Electroencephalography (EEG) is the patient-side test providing an objective diagnostic criterion for seizures and epilepsy. Despite this, EEG is thought to be rarely used in veterinary neurology. Objectives: This survey study aims to better understand the current canine EEG usage and techniques and barriers in veterinary neurology. Methods: The online Qualtrics link was distributed via listserv to members of the American College of Veterinary Internal Medicine (ACVIM) Neurology Specialty and the European College of Veterinary Neurology (ECVN), reaching at least 517 veterinary neurology specialists and trainees worldwide. Results: The survey received a 35% response rate, for a total of 180 participant responses. Fewer than 50% of veterinary neurologists are currently performing EEG and it is performed infrequently. The most common indication was to determine a discrete event diagnosis. Other reasons included monitoring treatment, determining brain death, identifying the type of seizure or epilepsy, localizing foci, sleep disorders, for research purposes, and post-op brain surgery monitorization. Most respondents interpreted their own EEGs. Clinical barriers to the performance of EEG in dogs were mainly equipment availability, insufficient cases, and financial costs to clients. Conclusion: This survey provides an update on EEG usage and techniques for dogs, identifying commonalities of technique and areas for development as a potential basis for harmonization of canine EEG techniques. A validated and standardized canine EEG protocol is hoped to improve the diagnosis and treatment of canine epilepsy.

Mechanistic Target of Rapamycin Inhibition Prevents Coronary Artery Remodeling in a Murine Model of Kawasaki Disease.

OBJECTIVE: Remodeling of the coronary arteries is a common feature in severe cases of Kawasaki disease (KD). This pathology is driven by the dysregulated proliferation of vascular fibroblasts, which can lead to coronary artery aneurysms, stenosis, and myocardial ischemia. We undertook this study to investigate whether inhibiting fibroblast proliferation might be an effective therapeutic strategy to prevent coronary artery remodeling in KD. METHOD: We used a murine model of KD (induced by the injection of the Candida albicans water-soluble complex [CAWS]) and analyzed patient samples to evaluate potential antifibrotic therapies for KD. RESULTS: We identified the mechanistic target of rapamycin (mTOR) pathway as a potential therapeutic target in KD. The mTOR inhibitor rapamycin potently inhibited cardiac fibroblast proliferation in vitro, and vascular fibroblasts up-regulated mTOR kinase signaling in vivo in the CAWS mouse model of KD. We evaluated the in vivo efficacy of mTOR inhibition and found that the therapeutic administration of rapamycin reduced vascular fibrosis and intimal hyperplasia of the coronary arteries in CAWS-injected mice. Furthermore, the analysis of cardiac tissue from KD fatalities revealed that vascular fibroblasts localizing with inflamed coronary arteries up-regulate mTOR signaling, confirming that the mTOR pathway is active in human KD. CONCLUSION: Our findings demonstrate that mTOR signaling contributes to coronary artery remodeling in KD, and that targeting this pathway offers a potential therapeutic strategy to prevent or restrict this pathology in high-risk KD patients.