Prior studies have failed to detect a convincing association between histologic lesions of inflammation and clinical activity in dogs with inflammatory bowel disease (IBD). We hypothesized that use of a simplified histopathologic scoring system would improve the consistency of interpretation among pathologists when describing histologic lesions of gastrointestinal inflammation. Our aim was to evaluate the correlation of histopathologic changes to clinical activity in dogs with IBD using this new system. Forty-two dogs with IBD and 19 healthy control dogs were enrolled in this retrospective study. Endoscopic biopsies from the stomach, duodenum, ileum, and colon were independently scored by 8 pathologists. Clinical disease activity was scored using the Canine Inflammatory Bowel Disease Activity Index (CIBDAI) or the Canine Chronic Enteropathy Clinical Activity Index (CCECAI), depending on the individual study center. Summative histopathological scores and clinical activity were calculated for each tissue (stomach, duodenum, ileum, and colon) and each tissue histologic score (inflammatory/morphologic feature). The correlation between CCECAI/CIBDAI and summative histopathologic score was significant (P < .05) for duodenum (r = 0.42) and colon (r = 0.33). In evaluating the relationship between histopathologic scores and clinical activity, significant (P < .05) correlations were observed for crypt dilation (r = 0.42), lamina propria (LP) lymphocytes (r = 0.40), LP neutrophils (r = 0.45), mucosal fibrosis (r = 0.47), lacteal dilation (r = 0.39), and villus stunting (r = 0.43). Compared to earlier grading schemes, the simplified scoring system shows improved utility in correlating histopathologic features (both summative histology scores and select histologic scores) to IBD clinical activity.

Significant interobserver variability in the diagnostic interpretation of endoscopic gastrointestinal (GI) specimens exists even with the use of World Small Animal Veterinary Association (WSAVA) standardization criteria. Chi-square analyses compared the extent of pathologists' agreement for microarchitectural features of inflammation in endoscopic specimens obtained from 253 animals of the original WSAVA study. Patterns of agreement between pathologists were classified as broad (3/4 pathologists agreed), dichotomous (2/4 pathologists agreed), or divergent (no agreement between pathologists). The simplified model for GI inflammation was based on those parameters for which the pathologists had either broad or minimally divergent opinions of histopathologic significance. In this model, the parameters chosen were as follows: gastric parameters (intraepithelial lymphocytes [IELs], lamina propria [LP] infiltrates, and mucosal fibrosis), duodenal parameters (villus atrophy, epithelial injury, IELs, crypt changes, and LP infiltrates), and colonic parameters (epithelial injury, crypt dilation, fibrosis, LP infiltrates, and goblet cell depletion). Preliminary data using this simplified model showed excellent correlation between pathologists in defining the presence and extent of GI inflammation in dogs.

OBJECTIVE:In the dog biopsy samples from the gastro-esophageal junction (GEJ) are rarely obtained during routine gastroscopy. The aim of this pilot study was to assess the histological quality of endoscopic biopsies sampled from the canine esophagus and cardia. It was hypothesised that it is possible to sample adequate specimens from these sites. MATERIALS AND METHODS:For this purpose 10 dogs with an indication for gastroscopy were enrolled in a prospective study. Biopsy samples were obtained with standard biopsy forceps for single use exactly from the GEJ thus containing preferably columnar epithelium from the cardia and squamous epithelium from the esophagus, respectively. In every dog the specimens were examined for size, layers and site, respectively. Study endpoint was reached when specimens originated from cardia and esophagus, showing at least epithelium and lamina propria mucosae, and a diameter >2mm on the slide, respectively. RESULTS:72 biopsy specimens (median 7, range 5-10) obtained from the GEJ were examined in 10 dogs. Specimens from the esophagus containing squamous epithelium with lamina propria mucosae were found in 5 of 10 (50.0%) dogs. Specimens from the cardia containing columnar epithelium with lamina propria mucosae were found in 10 of 10 (100.0%) dogs. Four of 10 (40.0%), and 10 of 10 (100.0%) dogs showed at least one specimen >2mm on the slide originating from the esophagus, and from the cardia, respectively. Histological quality was found to be adequate in 4 of 10 (40.0%) dogs, showing specimens of adequate size, originating from both esophagus and cardia, and containing at least epithelium and lamina propria mucosae. CONCLUSION AND CLINICAL RELEVANCE:The pilot study provides evidence that during routine gastroscopy it is possible to sample endoscopic biopsies from the cardia and with limitations from the esophagus showing a quality adequate for histological examination of the epithelium and the lamina propria mucosae.

Objective: To prove the hypothesis that a polyneuropathy in Alaskan Malamutes has a genetic background. Material and methods: Pedigrees of 131 related Alaskan Malamutes were included in the current study. Neurological examination, electrodiagnosis as well as muscle and nerve biopsies could be performed in 10 dogs. Information about the disease status of the other 121 Alaskan Malamutes were supplied by referring veterinarians, breeders and owners. Segregation analysis using four different models (monogenic, polygenic, mixed monogenic-polygenic and the phenotypic model) was performed on 71 dogs to test the different mechanisms of genetic transmission. Results: In seven clinically affected dogs abnormal electromyographic findings and reduced nerve conduction velocity were detected. Suspected diagnosis of polyneuropathy was confirmed by nerve biopsy results, characterized by axonal degeneration and hypomyelination. Muscle specimens revealed signs of neurogenic myopathy. Three related clinically normal Alaskan Malamutes also displayed moderate neuromuscular changes in histopathology. In the segregation analysis the polygenic model proved as best suitable to explain the observed segregation pattern among all other models tested. Conclusion: The current study could demonstrate that polyneuropathy in Alaskan Malamutes is a hereditary disease with variable phenotypic expression ranging from severely affected to subclinical forms, which has to be considered in future gene analysis studies.

BACKGROUND:Prior studies failed to detect significant association between hypoalbuminemia and small intestinal lesions.HYPOTHESIS:Use of pictorial templates will enhance consistency of interpathologist interpretation and identification of intestinal lesions associated with hypoalbuminemia.ANIMALS:Tissues from 62 dogs and 25 cats examined as clinical cases at 7 referral veterinary practices in 4 countries.METHODS:Retrospective, observational study. Histopathology slides from sequential cases undergoing endoscopic biopsy were examined by 4 pathologists by pictorial templates. Changes for 9 microscopic features were recorded as normal, mild, moderate or severe, and 2- and 4-point scales were tested for consistency of interpretation. Logistic regression models determined odds ratios (OR) of histologic lesions being associated with hypoalbuminemia while kappa statistics determined agreement between pathologists on histologic lesions.RESULTS:There was poor agreement (kappa = -0.013 to 0.3) between pathologists, and institution of origin of slides had effect (kappa = 1.0 for 3 of 4 lesions on slides from Institution 5) on agreement between pathologists on selected histologic features. Using 2 point as opposed to 4-point grading scale increased agreement between pathologists (maximum kappa = 0.69 using 4-point scale versus maximum kappa = 1.0 using 2-point scale). Significant association (P = .019- .04; 95% OR = 3.14-10.84) between lacteal dilation and hypoalbuminemia was found by 3 pathologists.CONCLUSIONS AND CLINICAL IMPORTANCE:Substantial inconsistency between pathologists remains despite use of pictorial template because of differences in slide processing. Distinguishing between mild and moderate lesions might be important source of the disagreement among pathologists.

A 3-year-old, male domestic shorthair cat was presented with signs of progressive muscular hypertrophy.A diagnosis of hypertrophic feline muscular dystrophy was made on the basis of the typical clinical presentation in cats and on the basis of the results of medical imaging, the electrodiagnostics and the findings on histopathology and immunohistochemistry of muscle biopsies.The cat was initially successfully treated with prednisolone.Eight months after presentation, however, the cat was euthanized because of acute renal failure.To the author's knowledge, this is the first report of hypertrophic feline muscular dystrophy in a domestic cat in Belgium.

Background: Chronic pulmonary diseases (recurrent airway obstruction [RAO]) have been reported to alter skeletal muscle cells in humans. The purpose of this study was to evaluate a potential relationship between pulmonary and muscle variables in horses with a clinical diagnosis of RAO. Muscle biopsies from healthy horses and from horses with RAO were investigated and the relationship between the severity of lung disease and the degree of muscular changes was determined.Hypothesis: We hypothesized that chronic pulmonary disease can lead to changes of the skeletal muscle in horses.Animals: Fifteen healthy horses (control) and 50 horses with RAO were examined.Methods: In a prospective clinical trial, a complete lung examination was performed in all horses. In all horses, muscle enzyme activity at rest and after exercise and muscle biopsies from the M. gluteus medius were examined.Results: None of the horses had clinical or histologic signs of primary or neurogenic myopathies. According to the clinical, endoscopic, and radiographic findings and with a scoring system, the horses with RAO were grouped according to the severity of pulmonary findings (15 horses mild, 24 horses moderate, 11 horses severe RAO). Pathologic changes of the skeletal muscle (fiber atrophy or fiber hypertrophy, myofibrillar degeneration, hyperplasia of mitochondria, and ragged-red-like fibers) were identified in most horses with RAO but in only a few individual control horses. In addition, a marked depletion of muscle glycogen storage was evident in the RAO horses but not in the control group. Other pathologic changes of skeletal muscle such as centralized nuclei and regenerating fibers were rare, but were more frequent in horses with lung diseases than in the control group. The degree of muscle cell changes was also graded with a scoring system and correlated with the severity of pulmonary disease (r = 0.55).Conclusion: Chronic pulmonary disease in horses is associated with structural changes in skeletal muscle.Clinical Importance: Because chronic pulmonary disease may affect muscles, early and effective therapy may prevent these changes. This finding could be of clinical importance but requires further studies.

The characterization of inflammatory change in endoscopic biopsy samples of the gastrointestinal mucosa is an increasingly important component in the diagnosis and management of canine and feline gastrointestinal disease. Interpretation has hitherto been limited by the lack of standard criteria that define morphological and inflammatory features, and the absence of such standardization has made it difficult, if not impossible, to compare results of retrospective or prospective studies. The World Small Animal Veterinary Association (WSAVA) Gastrointestinal Standardization Group was established, in part, to develop endoscopic and microscopical standards in small animal gastroenterology. This monograph presents a standardized pictorial and textual template of the major histopathological changes that occur in inflammatory disease of the canine and feline gastric body, gastric antrum, duodenum and colon. Additionally, a series of standard histopathological reporting forms is proposed, to encourage evaluation of biopsy samples in a systematic fashion. The Standardization Group believes that the international acceptance of these standard templates will advance the study of gastrointestinal disease in individual small companion animals as well as investigations that compare populations of animals.

Purpose: Tumor derived exosomes isolated from malignant effusions could serve as a new source of tumor-rejection antigens, and thus may lead to a new way to tumor immunotherapy. Exosomes are membrane vesicles of 60 to 100 nm in diameter with a density of 1.21 g/ml, of endocytic origin, and are released in vitro by tumor cell lines in culture supernatants. In vivo and in vitro experiments suggest that exosomes are involved in the transfer of tumor antigens to antigen presenting cells. The aim of this study was the verification of the presence of exosomes in malignant effusions of gynecological cancer patients and their subsequent isolation and characterization. Methods: The isolation of exosomes was performed by ultracentrifugation on sucrose gradients of eight malignant effusions of patients with breast cancer, ovarian cancer, endometrial cancer and pancreatic cancer. The characterization of the exosomes was performed using SDS-PAGE with Coomassie blue staining, Western blot analysis and electron microscopy. Results: In malignant effusions of gynecological patients membrane vesicles with a diameter of 60-100 nm could be detected. These isolated membrane vesicles have antigen-presenting molecules (MHC class-1) on their surface. These criteria and the morphological parameters (electron microscopy) indicate the similarities between these vesicles and the exosomes. Conclusion: Exosomes accumulate in malignant effusions of patients with cancer. Importantly, we were able to identify exosomes from malignant effusions of patients with endometrial and pancreatic cancer, which has not been described heretofore. Further investigations using these exosomes have to be performed to analyze the induction of the tumor specifity of T cells when pulsed with dendritic cells.

Fragestellung: Aus malignen Pleuraergüssen oder malignem Aszitis isolierte Exosomen könnten als natürliche Quelle von Tumorantigenen dienen und somit in der Immuntherapie gynäkologischer Tumoren eine vielversprechende Alternative zu tumorzellbasierten Vakzinen darstellen. Exosomen sind Membranvesikel endosomalen Urprungs mit einer Größe von 60–100 nm und einer Dichte von 1,21g/ml. Die Bildung dieser Membranvesikel durch Tumorzellen konnte in erster Linie in vitro gezeigt werden. Ziel unserer Untersuchung war der Nachweis von Exosomen in vivo im malignen Aszites/Pleuraerguß beim Mammakarzinom und gynäkologischen Tumoren.

Fragestellung: Exosomen aus malignem Aszites und Pleuraerguss bieten möglicherweise eine natürliche und neue Quelle der Gewinnung von Exosomen und stellen somit eine Erfolg versprechende Alternative zu tumorzellbasierten Vakzinen dar. Exosomen sind Membranvesikel endosomalen Urprungs mit einer Größe von 60 - 100 nm und einer Dichte von 1,21 g/ml. Die Bildung dieser Membranvesikel durch Tumorzellen konnte in erster Linie in vitro gezeigt werden. Ziel unserer Untersuchung war der Nachweis von Exosomen in vivo im malignen Aszites/Pleuraerguss beim Mammakarzinom und gynäkologischen Tumoren. Methode: Maligner Aszites von gynäkologischen Tumoren und maligner Pleuraerguss beim Mammakarzinom wurde bei acht Patientinnen mittels differenzieller Zentrifugation in fünf Schritten angereichert, gefolgt von einer Dichtezentrifugation mit kontinuierlichem Sukrosegradienten in einer Ultrazentrifuge. Die Charakterisierung und Identifizierung der Exosomen erfolgte mittels SDS-PAGE mit Coomassie-blue-Proteinfärbung, Westernblot-Analyse und Elektronenmikroskopie. Ergebnisse: Exosomen konnten im malignen Pleuraerguss und malignen Aszites gynäkologischer Patientinnen elektronenmikroskopisch nachgewiesen werden. Schlussfolgerung: Wir berichten erstmalig über den Nachweis von Exosomen aus malignem Aszites beim Endometrium- und Pankreaskarzinom. Exosomen bieten möglicherweise eine natürliche und neue pluripotente Quelle für Tumorantigene und eröffnen somit einen neuen Weg der Immunstrategie.

Muscle & NerveVolume 34, Issue 1 p. 122-122 Letters to the Editor Evidence for MHC-1–restricted CD8+ T-cell–mediated immunopathology in canine masticatory muscle myositis and polymyositis G. Diane Shelton DVM, PhD, G. Diane Shelton DVM, PhD Department of Pathology, University of California at San Diego, La Jolla, California, USASearch for more papers by this authorOrlando Paciello DVM, PhD, Orlando Paciello DVM, PhD Department of Pathology and Animal Health, University of Naples Federico II, Naples, ItalySearch for more papers by this author G. Diane Shelton DVM, PhD, G. Diane Shelton DVM, PhD Department of Pathology, University of California at San Diego, La Jolla, California, USASearch for more papers by this authorOrlando Paciello DVM, PhD, Orlando Paciello DVM, PhD Department of Pathology and Animal Health, University of Naples Federico II, Naples, ItalySearch for more papers by this author First published: 31 March 2006 https://doi.org/10.1002/mus.20547Citations: 5Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References 1 Dalakas MC. Inflammatory disorders of muscle: progress in polymyositis, dermatomyostis and inclusion body myositis. Curr Opin Neurol 2004: 17: 561–567. 10.1097/00019052-200410000-00006 CASPubMedWeb of Science®Google Scholar 2 Engel AG, Hohlfeld R. Inflammatory myopathies. The polymyositis and dermatomyositis syndromes. In: AG Engel, C Franzini-Armstrong, editors. Myology. New York: McGraw-Hill; 2004, p 1321–1388. Google Scholar 3 Evans J, Levesque, D, Shelton GD. Canine inflammatory myopathies: a clinicopathologic review of 200 cases. J Vet Intern Med 2004; 18: 679–691. 10.1111/j.1939-1676.2004.tb02606.x CASPubMedWeb of Science®Google Scholar 4 Neumann J, Bilzer T. Evidence for MHC-1–restricted CD8+ T-cell–mediated immunopathology in canine masticatory muscle myositis and polymyositis. Muscle Nerve 2006; 33: 215–224. 10.1002/mus.20456 CASPubMedWeb of Science®Google Scholar 5 Pumarola M, Moore PF, Shelton GD. Canine inflammatory myopathy: analysis of cellular infiltrates. Muscle Nerve 2004; 29: 782–789. 10.1002/mus.20043 CASPubMedWeb of Science®Google Scholar 6 Shelton GD, Cardinet GH, Bandman E. Canine masticatory muscle disorders: a study of 29 cases. Muscle Nerve 1987; 10: 753–766. 10.1002/mus.880100812 CASPubMedWeb of Science®Google Scholar Citing Literature Volume34, Issue1July 2006Pages 122-122 ReferencesRelatedInformation

Masticatory muscle myositis (MMM) is the most common inflammatory myopathy (IM) in dogs, associated with antibodies against myosin. To further elucidate the immunopathogenesis, we investigated muscles of 53 dogs with MMM, 32 dogs with polymyositis (PM), and 4 dogs suffering from both, with regard to the presence and location of CD4(+) and CD8(+)T cells, B cells, macrophages, major histocompatibility complex (MHC) class I and class II antigens, and autoantibodies. CD8(+)T cells were found in MMM (91%) and PM (75%), mostly paralleled (68% and 61%) by enhanced expression of MHC class I antigen on muscle fibers. CD8(+)T cells invading intact and neighboring necrotic muscle fibers were present in MMM (39%) and PM (42%). Dogs with MMM lacking intramuscular (26%) and circulating (36%) autoantibodies also had CD8(+) T-cell infiltrations and muscle-fiber lesions. Since MHC class I antigen and CD8(+) T cells were detected in the presence of CD4(+) T cells, regardless of antimuscular antibodies, we consider MMM and PM in the dog as a CD8(+) T-cell-mediated immunopathological disease that initiates muscle-fiber destruction and leads to production of myosin autoantibodies.

The human glioblastoma-derived cell lines 86HG-39, 87HG-28 and 87HG-31, used for the production of monoclonal antibodies (mAbs) against glioma-associated antigens (GAA), were characterized in terms of morphology, growth behaviour, chromosomes and antigen expression. In the primary tumours, differential expression of glial fibrillary acidic protein, S100 protein, Leu-7 and GAA as defined by mAbs MUC 2-39, MUC 2-63 and MUC 8-22 was demonstrated. Receptors for epidermal growth factor (EGFr) and nerve growth factor (NGFr) were found in many cells in short-term cultures, but the transferrin receptor (Tr) was found in only a few cells of 87HG-28. In permanent cell lines, differentiation antigens and EGFr decreased and Tr increased markedly. NGFr and GAA remained stable. Transplantation tumours of 86HG-39 were partly positive for Tr and GAA. Chromosomal analysis revealed that the 86HG-39 and 87HG-28 cell lines had a hypodiploid or diploid stem line with lines in the hypotetraploid to tetraploid region for 50 in vitro passages. The 87HG-31 cell line had chromosomal patterns in the hypotriploid to triploid region. A gain of chromosomes was seen in the groups C7, C8, C10, D14, F19, F20, G21, G22. The variability of antigens in these tumours and especially during long-term cultivation probably reveals an ability to influence the growth of malignant glioma cells via the respective effector molecules.