An osteoid variant of cutaneous melanoma in a dog detected by S100 and melan a markers

Osteoid malignant melanoma is a rare type of melanoma described in humans and dogs with some areas of bone differentiation. In this tumour, the origin of the bone matrix remains unclear. We report one case of this variant with, for the first time, a cutaneous origin in a dog. Malignant melanomas are aggressive tumours. Amelanotic tumours are sometimes difficult to recognize as they require immunohistochemical evaluation for an adequate diagnosis and we have used anti-vimentin, S100, and melan A antibodies for identification. Melan A is less sensitive but more specific than S100 in identifying amelanotic melanomas. This tumour was positive for vimentin, S100 and melan A, including the areas of osteoid. These results suggest osteoid differentiation of tumour cells rather than induced stromal metaplasia.     

Aetiology of canine otitis externa: a retrospective study of 100 cases

The objective of this retrospective study was to investigate in 100 dogs with otitis externa (OE) the possible associations between signalment, history, clinical and laboratory findings and the various primary, secondary and perpetuating causative factors of ear canal inflammation. The age of the dogs ranged from 3 months to 14 years (median: 4.75 years) and they included 45 males and 55 females. Cocker spaniels, Jura des Alpes and Brittany spaniels were significantly overrepresented among dogs with OE when compared to the hospital canine population. In the majority of the cases, OE was chronic-recurrent (63%) or bilateral (93%). Allergic dermatitis (43/100 dogs), grass awns (12/100) and otoacariasis (7/100) were the most common primary causative factors; no primary factor could be incriminated in 32 cases and more than one was found in three dogs. Malassezia spp. (66/100 dogs), cocci (38/100) and rods (22/100) were the secondary causative factors, while ear canal stenosis (38/100) and tympanic membrane perforation-otitis media (25/100) were the most important perpetuating factors. Atopic dermatitis and adverse food reactions-associated OE was more common in females and dogs with a history of pruritic skin disease, while grass awn-induced OE occurred in cocker spaniels and acute cases. Tympanic membrane perforation was less frequent in atopic dermatitis and adverse food reactions-associated OE, but more common when otoscopic and ear canal cytological examination revealed the presence of grass awns and rods, respectively. Finally, cocci overgrowth was positively associated with ear canal stenosis.     

Immunohistochemical staining for S100 protein in the diagnosis of canine amelanotic melanoma

Formalin-fixed, paraffin-embedded tissue samples of canine amelanotic melanomas and normal canine tissues were studied immunohistochemically for the presence of S100 protein. Use of the avidin-biotin complex procedure demonstrated variable amounts of S100 protein in the tumor cell cytoplasm and nuclei in 26 of 31 tumors. S100 protein was not observed in some other common canine skin tumors stained by the avidin-biotin complex technique. These were a mast cell tumor, fibrosarcoma, mammary gland adenocarcinoma, histiocytoma, transmissible venereal tumor, and a thyroid gland adenocarcinoma. Among normal tissues the presence of S100 protein was demonstrated in chondrocytes in the trachea, myoepithelial cells in the breast, melanocytes in the skin, some sweat glands and ducts in the skin, stellate cells in the pituitary, and interdigitating reticulum cells in the lymph node and in Peyer's patches. These results indicate that the avidin-biotin complex procedure for demonstrating S100 protein is a useful diagnostic tool in the diagnosis of canine amelanotic melanoma.     

A retrospective study of twenty-nine spinal tumours in the dog and cat

In a retrospective study of twenty-nine spinal tumours in dogs and cats approximately equal numbers of extradural and intradural tumours were found. The most prominent tumour type was the group of primary tumours of bone which included osteosarcomas, fibrosarcomas, hemangiosarcomas and multiple cartilagenous exostoses. Primary sarcomas of the spinal cord were the next most prominent type of tumour identified. A close clinicopathological correlation was found for tumour localization; however, intradural and extradural tumours could not be differentiated clinically. In comparison with extramedullary tumours which were evenly divided between acute and insidious onset, the majority of animals with intramedullary tumours had acute onset of neurological signs. In general, the intramedullary tumours had the shortest duration of clinical signs (1.7 weeks) whereas the extradural tumours had a mean duration of 3.4 weeks and intradural-extramedullary tumours had the longest mean duration (5.7 weeks). Contrast radiography was the most useful diagnostic aid. It confirmed the regional localization in twenty-two of twenty-four animals     

Expression and significance of p53, rb,p21/waf-1, p16/ink-4a,and PTEN tumor suppressors in canine melanoma

The role of tumor suppressor genes in the pathogenesis of canine melanoma is incompletely understood. The genes encoding the tumor suppressors p53, Rb, p21 (waf-1), p16 (ink-4a), and PTEN have been postulated to contribute to the pathogenesis of melanoma in humans and experimental animal models. To assess whether inactivation of these genes similarly contributes to the origin and progression of canine melanoma, we examined their expression in seven distinct canine melanoma cell lines and in 31 retrospective samples (representing 29 dogs) of spontaneous canine melanoma. Various patterns suggestive of loss of tumor suppressor function emerged in these cell lines. The most frequently observed abnormality was loss or significant reduction of p16 expression in six of seven cell lines and in 21 of 26 tumor samples. Loss or significant reduction of PTEN expression was seen in four of seven cell lines and in 13 of 27 tumor samples. Although p53 was detectable in all the cell lines and in 24 of 30 tumors, exclusion of p53 from the nuclear compartment was observed in each of the cell lines and in 18 of 25 tumor samples. These results indicate that loss of function of these tumor suppressor proteins is a common occurrence that may contribute to the origin of canine melanoma. In our sample population, abnormalities in the expression or localization of one or more tumor suppressor proteins occurred with similar frequency in malignant and benign tumors; thus, additional work is necessary to determine how these proteins may impact disease progression and response to therapy.     

Absence of S100A4 (mts1) gene mutations in various canine and feline tumours. Detection of a polymorphism in feline S100A4 (mts1)

Ninety canine and 101 feline tumours of various types were investigated for gene mutations in the coding regions of the metastasis-associated gene S100A4 (mts1). No gene mutations were present in the analysed genomic area. A widespread histidine/tyrosine polymorphism was detected in codon 17 of S100A4.     

A retrospective study of diagnosis in 109 cases of canine lower respiratory disease

Between 1983 and 1988, 109 cases of canine lower respiratory tract disease were examined and categorised according to clinical, radiographic and bronchoscopic findings and cytology of bronchial mucus. Non-specific chronic tracheobronchitis was diagnosed in 19 cases, bronchopneumonia (subacute or chronic) in 12 cases, bronchiectasis in 10 cases, parasitic bronchitis due to Oslerus osleri in 20 cases, eosinophilic bronchitis with or without bronchiectasis or alveolar infiltration in 25 cases, bronchial foreign bodies, mainly cereal ears, in 14 cases, primary neoplasia (adenocarcinomas) in six cases and miscellaneous disease in three cases. Features of these conditions are discussed. Although the aetiology of chronic bronchial disease may not always be ascertained, it is important to assign a dog to a diagnostic category so that the owner can be given a more accurate prognosis.     

Expression of the MDR1 gene and P-glycoprotein in canine mast cell tumor cell lines

Cellular drug resistance to antineoplastic drugs is often due to the presence of a drug efflux pump that reduces intracellular drug accumulation and chemosensitivity. P-glycoprotein (P-gp), which is encoded by the MDR1 gene, is considered to function as an ATP-driven membrane drug efflux pump and appears to play an important role in tumor cell resistance. In the present report, we assessed the expression of MDR1 by RT-PCR in three canine mast cell tumor cell lines, TiMC, CoMS and LuMC, originating from a cutaneous tumor, an oral-mucosal tumor and a gastrointestinal tumor, respectively. P-gp expression was also examined by Western blot analysis, while the functional activity of P-gp was assessed by flowcytometric analysis of intracellular rhodamine-123 (Rhd-123) uptake. The results revealed that MDR1 gene and P-gp were both expressed in CoMS and LuMC cells, whereas neither was present in TiMC cells. In CoMS and LuMC cells, intracellular uptake of Rhd-123 increased in the presence of verapamil, a functional modulator of P-gp. In contrast, TiMC cells did not show any changes in the intracellular accumulation of Rhd-123 after the verapamil addition. These findings suggest that the expressions of MDR1 gene and P-gp probably contribute to cellular drug resistance in canine mast cell tumors.     

Contrast radiographic findings in canine bacterial discospondylitis: a multicenter, retrospective study of 27 cases

A multicenter, retrospective study was undertaken to evaluate contrast radiographic findings in canine bacterial discospondylitis. Records and myelograms or epidurograms of 27 patients were obtained from five colleges of veterinary medicine. Fifteen cases (56%) were evaluated as having some degree of spinal cord compression. The majority (73.3%) of the cases had only soft tissue as the compressive mass. The median compression for all cases was 5% of the vertebral canal. No difference was noted for compression based on anatomical site (i.e., cervical versus thoracolumbar versus lumbosacral). No significant correlation between degree of lesion compression and clinical outcome was noted, but there was a trend toward increased mortality with greater compression. There was no correlation between the ambulatory status and the ultimate outcome. Three of the 15 (20%) cases showed vertebral subluxation. Results of this study indicate that static spinal cord compression is not a significant component of the neurological dysfunction associated with bacterial discospondylitis. Identification of vertebral subluxation in some patients may indicate a dynamic lesion that should be evaluated with stress radiography.     

Expression of a tumor-associated antigen, RCAS1, in canine mammary tumors

Receptor-binding cancer antigen expressed on SiSo cells (RCAS1), one of novel cancer cell-surface antigens, is strongly expressed in invasive cancers. RCAS1 inhibits the in vitro growth of lymphocytes such as T cells and natural killer (NK) cells, and induces apoptotic cell death. We investigated the expression of RCAS1 in canine mammary tumor cell lines and tumor cells by immunohistochemistry, and also in situ deoxyribonucleic acid (DNA) fragmentation in tumor-infiltrating lymphocytes (TILs) by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method. All canine mammary tumor cell lines expressed RCAS1 at both the messenger ribonucleic acid (mRNA) and protein level. Immunohistochemically, RCAS1 was negative in 100% of normal mammary glands, but was expressed in 100% of malignant tumors examined. In most malignant mammary tumors, RCAS1 was localized in the cytoplasm with no polarity of expression. In benign mammary tumors, it was detected on the luminal surface of the tumor cell. RCAS1 expression or localization was significantly correlated with malignancy. In situ DNA fragmentation of CD3-positive TILs was observed in RCAS1-expressing tumors. RCAS1-expressing tumors, indicating a possible induction of apoptotic cell death in TILs through RCAS1 expression. These observations suggest that RCAS1 probably plays an important role in tumor progression and escape from immune surveillance in canine mammary tumors.     

Epidemiology and clinical significance of canine distichiasis: A retrospective study of 291 cases

Objective

To describe the epidemiological factors and clinical significance of canine distichiasis.

Animals studied

Two hundred and ninety-one client-owned dogs.

Methods

Retrospective study of medical records for canine patients diagnosed with distichiasis between 2010 and 2019 in an ophthalmology specialty practice. The breed, sex, skull conformation, coat type, age at the time of diagnosis, reason for presentation, clinical examination findings, and affected eyelid(s) were reviewed.

Results

The prevalence of distichiasis was 5.5% (95% confidence interval (CI): 4.9–6.1) in the population of dogs presented to an ophthalmology specialty practice. The breeds with the highest prevalence were English bulldogs (35.2%, 95% CI: 26.7–43.7) and American cocker spaniels (19.4%, 95% CI: 8.3–30.5). The prevalence was significantly higher in brachycephalic dogs (11.9%, 95% CI: 9.8–14.0) than in non-brachycephalic dogs (4.6%, 95% CI: 4.0–5.3) and in short-haired dogs (8.2%, 95% CI: 6.8–9.6) than in dogs with other coat types (5.3%, 95% CI: 4.5–6.1). Most dogs were affected bilaterally (63.6%, 95% CI: 58.0–69.1). Among dogs with clinical signs, 39.0% (95% CI: 26.5–51.4) exhibited corneal ulceration, including superficial ulcers (28.8%, 95% CI: 17.3–40.4) and deep stromal ulcers (10.2%, 95% CI: 2.5–17.8). Distichiasis was non-irritating in 85.0% (95% CI: 80.6–89.4) of affected dogs.

Conclusion

This study reports the largest cohort of canine distichiasis to date. In a large proportion of dogs, distichiasis was a non-irritating condition. However, brachycephalic breeds, especially English bulldogs, were the most frequently and severely affected.     

Antitumor effects of celecoxib in COX-2 expressing and non-expressing canine melanoma cell lines

Cyclooxygenase-2 (COX-2) is a potential target for chemoprevention and cancer therapy. Celecoxib, a selective COX-2 inhibitor, inhibits cell growth of various types of human cancer including malignant melanoma. In dogs, oral malignant melanoma represents the most common oral tumor and is often a fatal disease. Therefore, there is a desperate need to develop additional therapeutic strategies. The purpose of this study was to investigate the anticancer effects of celecoxib on canine malignant melanoma cell lines that express varying levels of COX-2. Celecoxib induced a significant anti-proliferative effect in both LMeC and CMeC-1 cells. In the CMeC cells, treatment of 50 μM celecoxib caused an increase in cells in the G0/G1 and a decreased proportion of cells in G-2 phase. In the LMeC cells, 50 μM of celecoxib led to an increase in the percentage of cells in the sub-G1 phase and a significant activation of caspase-3 when compared to CMeC-1 cells. In conclusion, these results demonstrate that celecoxib exhibits antitumor effects on canine melanoma LMeC and CMeC-1 cells by induction of G₁-S cell cycle arrest and apoptosis. Our data suggest that celecoxib might be effective as a chemotherapeutic agent against canine malignant melanoma.     

Effect of transforming growth factor-beta1, insulin-like growth factor-I,and hepatocyte growth factor on proteoglycan production and regulation in canine melanoma cell lines

OBJECTIVE:To identify extracellular proteoglycans produced by canine melanoma cell lines and analyze the effect of transforming growth factor-beta1 (TGF-beta1), insulin-like growth factor-I (IGF-I), and hepatocyte growth factor (HGF) on these proteoglycans. SAMPLE POPULATION: 3 canine melanoma cell lines (ie, CML-1, CML-6M, and CML-10c2). PROCEDURE: Extracellular proteoglycans were analyzed by use of metabolic labeling and western immunoblot analysis. The effect of TGF-beta1 on cell proliferation was determined by incorporation of 5-bromo-2'-deoxyuridine. RESULTS: The CML-1 and CML-6M melanoma cell lines produced 2 main extracellular proteoglycans. One of them was identified as versican, a proteoglycan found in undifferentiated human melanoma cell lines. The CML-10c2 cells produced a small amount of extracellular proteoglycans. Addition of TGF-beta1 (1.25 to 6.25 ng/ml) increased the release of sulfated proteoglycans into the medium. The TGF-beta1 had mainly a posttranslational effect, because it increased the molecular mass of the sulfated bands. Addition of IGF-I (50 ng/ml) slightly increased production of proteoglycans in the CML-6M cell line, whereas HGF (50 ng/ml) did not have any effect on proteoglycan production. CONCLUSIONS AND CLINICAL RELEVANCE: The proteoglycan content and response toTGF-beta1 treatment for CML-1 and CML-6M canine melanoma cell lines are similar to that for undifferentiated human melanoma cell lines. In contrast, CML-10c2 cells produced a low amount of proteoglycans with high molecular weight. Because these extracellular proteoglycans are involved in the control of cell adhesion, proliferation, and migration, they may play an important role in the progression of melanomas in dogs.     

Metabolic and toxic causes of canine seizure disorders: A retrospective study of 96 cases

A wide variety of intoxications and abnormal metabolic conditions can lead to reactive seizures in dogs. Patient records of dogs suffering from seizure disorders (n = 877) were reviewed, and 96 cases were associated with an underlying metabolic or toxic aetiology. These included intoxications by various agents, hypoglycaemia, electrolyte disorders, hepatic encephalopathy, hypothyroidism, uraemic encephalopathy, hypoxia and hyperglycaemia. The incidence of the underlying diseases was determined.The most common causes of reactive seizures were intoxications (39%, 37 dogs) and hypoglycaemia (32%, 31 dogs). Hypocalcaemia was the most frequent electrolyte disorder causing reactive seizures (5%) and all five of these dogs had ionised calcium concentrations ?0.69 mmol/L. Eleven per cent of dogs with seizures had metabolic or toxic disorders and this relatively high frequency emphasises the importance of a careful clinical work-up of cases presented with seizures in order to reach a correct diagnosis and select appropriate treatment options.     

Melan A and S100 protein immunohistochemistry in feline melanomas: 48 cases

Immunohistochemistry, using a monoclonal antibody to Melan A and a polyclonal antibody to S100 protein, was applied to 48 formalin-fixed, paraffin-embedded specimens of feline melanoma. Forty-two cutaneous, three oral, one mucocutaneous, and two metastatic melanomas comprised the tumors. Thirty-two tumors (67%) were positive for Melan A and 42 (87.5%) were positive for S100. All but one of the tumors that were positive for Melan A were also positive for S100. S100 was detected in 11 of 16 tumors that were negative for Melan A. Seventy-five percent (9 of 12) of amelanotic melanomas were negative for Melan A. Normal adrenal cortex, the cerebellum, and the skin had cells that were positive for Melan A. Sebaceous adenoma was the only nonmelanocytic tumor examined that reacted with antibody to Melan A. Although less sensitive than S100 protein, Melan A is more specific for melanoma and is useful in differentiating feline cutaneous melanoma from the more common pigmented basal cell tumor.     

Epidemiology, clinical signs, histopathology and molecular characterization of canine leproid granuloma: a retrospective study of cases from Brazil

Thirty-eight cases of canine leproid granuloma were diagnosed between 2000 and 2008. Diagnosis was based upon clinical and histopathological findings and the presence of acid-fast bacilli in skin sections. The clinical lesions were localized predominantly on the pinnae and included papules, plaques and nodules, with or without ulceration. Boxer dogs were the breed most affected. Histopathological findings included nodular to diffuse pyogranulomatous, lymphoplasmocytic inflammatory infiltrates, with or without necrosis, localized in the dermis or subcutaneous tissue. The bacillary loading and morphology were variable among the lesions analysed. There was no significant correlation between bacterial load and histopathological pattern, dominant type of inflammatory infiltration or the amount of necrosis or giant cells. No correlation was observed between giant cells and histopathological pattern. In the majority of cases where a PCR-based assay was done, a novel mycobacterium species as the main aetiological agent was identified, as reported in previous studies.