Immunohistochemical expression of calretinin in canine testicular tumours and normal canine testicular tissue

Calretinin is a calcium-binding protein expressed abundantly in the central and peripheral neural tissues. It has been demonstrated to be a valuable marker in human testicular neoplasia. The immunohistochemical expression of calretinin has been studied in 102 samples of normal (n=25) and three different neoplastic canine testicular tumours (n=77). In normal canine testis, calretinin expression was restricted to Leydig and Sertoli cells of the testis. In tumour tissues, calretinin expression was detected in all tumours investigated (interstitial cell tumours, seminoma, and Sertoli cell tumours), with a cytoplasmic and nuclear pattern of cellular distribution. The present work reports, for the first time, calretinin immunohistochemical expression in normal and neoplastic canine testis.     

A canine peripheral nerve sheath tumor including peripheral nerve fibers

Peripheral nerve sheath tumor was found in a 7-year-old male mongrel dog. The tumors were located in the right cheek subcutis and oral submucosa. Histologically, neoplastic cells were arranged in streaming bundles, occasionally interlacing bundles or whorls of elongated and spindle cells. Cellular atypia was poor and mitotic figures were rarely observed. Ultrastructurally, neoplastic cells had basement membrane, typical of Schwann cells. One bundle of normal peripheral nerve fibers and some myelinated axons were seen within the tumor tissues. Immunohistochemically, neoplastic cells reacted to vimentin, glial fibrillary acidic protein, S-100 protein and neuron specific enolase. In addition to the above immunoreactions, the included nerve fibers were positive for myelin basic protein and neurofilament protein. This paper also discusses immunohistochemical findings on differential diagnosis in comparison with those of canine hemangiopericytomas reported hitherto.     

Morphologic and immunohistochemical features of two spontaneous peripheral nerve tumors in Wistar rats

Morphologic features and S-100 protein immunoreactivity of a benign and malignant peripheral nerve sheath tumor were studied in two Wistar rats. Neoplasms that developed in untreated control rats from tumor bioassays were S-100 protein positive and had similar histopathologic features. Each peripheral nerve sheath tumor was encapsulated and composed of spindle cells arranged around small thin-walled blood vessels. Palisaded tumor cells were in the benign peripheral nerve sheath tumor while cells of the malignant peripheral nerve sheath tumor had cellular atypia and moderate numbers of mitoses. Ultrastructural examination of the malignant peripheral nerve sheath tumor revealed cells with external lamina and interdigitation of cytoplasmic processes. Intracytoplasmic concentric lamellae were seen; they were regularly spaced with a periodicity of about 15 nm. Such structures, indistinguishable from myelin sheaths, have not been commonly associated with peripheral nerve sheath tumors in man. Electron microscopy and immunohistochemistry were useful in the diagnosis of these tumors as Schwannomas and in differentiation from other spindle cell tumors.     

Testicular Neoplasms (Interstitial and Sertoli Cell Tumours) in a Domestic Ferret (Mustela putorius furo)

Unilateral testicular enlargement was detected in a 5‐years‐old domestic ferret during a routine sterilization. The right testicle showed two different types of proliferative lesions: (i) round nodules, well demarcated, showing a soft yellow tissue; (ii) white nodules, firm, with irregular‐shaped invaginations. Microscopically, the neoplastic proliferations were identified as an interstitial neoplasm and Sertoli cell tumour, respectively. The left testicle was small and showed intense testicular atrophy. Clinical evaluation of the ferret did not show any other apparent pathological processes. This study is the first case reporting the concomitant occurrence of a Sertoli cells tumour and an interstitial cell tumour in a domestic ferret.     

Tumor-specific diagnostic marker for transmissible facial tumors of Tasmanian devils: immunohistochemistry studies

Devil facial tumor disease (DFTD) is a transmissible neoplasm that is threatening the survival of the Tasmanian devil. Genetic analyses have indicated that the disease is a peripheral nerve sheath neoplasm of Schwann cell origin. DFTD cells express genes characteristic of myelinating Schwann cells, and periaxin, a Schwann cell protein, has been proposed as a marker for the disease. Diagnosis of DFTD is currently based on histopathology, cytogenetics, and clinical appearance of the disease in affected animals. As devils are susceptible to a variety of neoplastic processes, a specific diagnostic test is required to differentiate DFTD from cancers of similar morphological appearance. This study presents a thorough examination of the expression of a set of Schwann cell and other neural crest markers in DFTD tumors and normal devil tissues. Samples from 20 primary DFTD tumors and 10 DFTD metastases were evaluated by immunohistochemistry for the expression of periaxin, S100 protein, peripheral myelin protein 22, nerve growth factor receptor, nestin, neuron specific enolase, chromogranin A, and myelin basic protein. Of these, periaxin was confirmed as the most sensitive and specific marker, labeling the majority of DFTD cells in 100% of primary DFTD tumors and DFTD metastases. In normal tissues, periaxin showed specificity for Schwann cells in peripheral nerve bundles. This marker was then evaluated in cultured devil Schwann cells, DFTD cell lines, and xenografted DFTD tumors. Periaxin expression was maintained in all these models, validating its utility as a diagnostic marker for the disease.     

Primary bilateral corneal nerve sheath neoplasm in a dog

A 15‐year‐old, neutered male, Shih Tzu cross developed progressive corneal stromal thickening and vascularization of the right eye, and 5 months later, of the left eye. Both eyes became blind due to extensive corneal opacification and were enucleated. Light microscopic examination revealed a diffuse corneal infiltrate of neoplastic mesenchymal cells, and immunohistochemistry revealed diffuse cytoplasmic vimentin immunoreactivity and variable cytoplasmic and nuclear immunoreactivity for S100 in the neoplastic cells. Transmission electron microscopy revealed desmosomes between contiguous cells, thread‐like cytoplasmic processes coated with basement membrane, extracellular bundles of collagen, and axonal degeneration consistent with features of a nerve sheath neoplasm. This is the first report of primary, bilateral corneal nerve sheath sarcoma in a canine.     

Ossifying fibroma in a miniature rex rabbit (Oryctolagus cuniculus)

In humans and animals, ossifying fibroma is a benign neoplasm that most frequently affects the mandible, often resulting in cosmetic deformities and malocclusion. It is considered rare in animals and most frequently affects young horses. A surgical biopsy of a solitary mass located beneath the gingiva in the right maxillary region, which had overgrown teeth and expanded the adjacent hard palate from a 6-year-old miniature Rex rabbit was submitted for light microscopic examination. The submitted incisional biopsy specimen was pale pink, firm, and nodular. Histopathologically, the neoplasm was composed of fibroblastic cells separated by abundant collagen. The neoplastic cells were interwoven with osteoblasts surrounding islands of mineralized, bony matrix containing few, widely spaced, often empty, lacunae. Minimal inflammation was present. Based on the histopathologic features, the tumor was diagnosed as an ossifying fibroma. To our knowledge, this is the first report of an ossifying fibroma in a rabbit.     

Peripheral neuroblastoma in a young labrador retriever

A 2-year-old Labrador Retriever developed atrophy of the right temporal muscle, subsequently showed generalized seizure and died 2 months after the clinical onset. Postmortem examination revealed the tumor masses in the right mandibulopharyngeal area, nasopharynx and intracranial space. Histopathologically, these tumor masses were composed of small round neoplastic cells and neuropil-like stroma separated by fibrovascular septa. In the neoplastic masses, small neoplastic cells with round to oval hyperchromatic nuclei and scanty cytoplasm predominated, and angulated neoplastic cells with larger nuclei and moderate cytoplasm were scattered. Immunohistochemically, neoplastic cells were positive for neuron specific enorase, neurofilament protein, chromogranin A, synaptophysin and tyrosine hydroxylase. Based on these findings, this case was diagnosed as peripheral neuroblastoma, presumably originated from the sympathetic ganglion, maybe right craninal cervical ganglion.     

Malignant peripheral nerve sheath tumour of the urinary bladder in a cat

A 14-year-old domestic shorthair cat presented with a 5-month history of urinary incontinence and inappro-priate elimination. Ultrasonography revealed a well-marginated, vascular mass of mixed echogenicity ex-tending from the dorsal wall of the urinary bladder into the lumen. Partial cystectomy was performed for re-moval of the urinary bladder mass; histopathological evaluation revealed a spindle cell neoplasm with a prominent palisading pattern. Histomorphologic features and immunohistochemical demonstration of vimentin, glial fibrillary acidic protein and S-100 protein, combined with negativity for smooth muscle actin and desmin were consistent with malignant peripheral nerve sheath tumour. This case report describes a novel location of malignant peripheral nerve sheath tumour; to the authors' knowledge, the bladder has not been described as a site of origin in the cat or any other domestic species.     

Malignant peripheral nerve sheath tumor as a cause of chronic cardiac insufficiency in cattle

Chronic cardiac insufficiency was associated with a malignant peripheral nerve sheath tumor in a cow. An eight-year-old cow developed a progressive condition (over a period of three months) characterized by an enhanced abdominal volume, reluctance to move, a positive jugular pulse, watery diarrhea and death. At necropsy, moderate subcutaneous edema and an enhanced hepatic lobular pattern were observed. A 23x20x11 cm firm, grayish-white mass adhered to and infiltrated the right atrium. Multiple firm, yellowish-white nodules of 0.5 to 12 cm in diameter were diffusely scattered in the epicardium and parietal pericardium. Histologically, the tumor was poorly circumscribed with foci of infiltration of the myocardium. The neoplastic cells had two major histologic patterns, Antoni types A and B. Within occasional foci, pleomorphic cells with an epithelioid appearance were present in addition to multinucleated cells with periodic acid Schiff (PAS)-positive cytoplasmic globules. Foci of cartilaginous and granular differentiations were interspersed among the neoplastic cells. Multiple vessels presented wall hyalinization and tumoral embolus. Large necrotic foci with mineralization and cholesterol clefts were also observed. Immunohistochemically, the tumor was positive for S100 protein, vimentin and neuron-specific enolase labeling.     

Primary malignant peripheral nerve sheath tumor with eosinophilic cytoplasmic globules arising from the greater omentum in a dog

A 10-year-old Golden Retriever dog had a solitary tumor mass arising from the greater omentum. Histologically, the tumor showed varying cellularity and patterns of cellular arrangement. In dense cellular areas, spindle-shaped cells were arranged in interlacing bundles. The sparse cellular area was characterized by loosely arranged fusiform cells. The neoplastic cells frequently contained PAS-positive eosinophilic globules in the cytoplasm, and mitotic figures were frequently observed. The tumor cells were positive to vimentin, S-100 protein, glial fibrillary acidic protein, myelin basic protein, neuron-specific enolase and myoglobin. The present tumor was diagnosed as a malignant peripheral nerve sheath tumor (MPNST) with eosinophilic cytoplasmic globules arising from the greater omentum. To our knowledge, this may be the first case of primary omental MPNST in domestic animals.     

Acute hindlimb paresis caused by extension of a peripheral nerve sheath tumour into the spinal canal of a horse

A 10-year-old Quarter Horse gelding presented for a large, slowly growing mass in the right thoracic region caudal to the withers (approximately at the level of thoracic vertebrae 10 to 16). The mass was surgically removed, the horse returned to normal riding activity, and no external regrowth was observed. Two years after surgery, the horse developed progressive signs of hindlimb weakness, ataxia and adopting a dog-sitting stance. Post-mortem examination confirmed spinal canal infiltration and spinal cord compression (at the level of the 12th thoracic vertebra) by a similar soft tissue sarcoma as diagnosed previously. Immunohistochemistry confirmed the tumour as a peripheral nerve sheath tumour (PNST) likely originating from the twelfth thoracic spinal nerve. This report demonstrates that with incomplete tumour resection, local recurrence and extension of a PNST into the central nervous system can occur even years after initial tumour removal. Immunohistochemistry to differentiate the type of soft tissue sarcoma in cases with close association to the nervous system may prove beneficial to anticipate this uncommon complication. Early surgical resection with adjunctive therapies should be considered in these cases.     

Subcutaneous Soft Tissue Sarcoma with Rhabdoid Features in a Dog

A nine-year-old male beagle dog had a white spherical mass in the subcutis of the left lumbar region. Microscopically, spindle to oval cells diffusely proliferated in the fibrous and myxoid stroma. Many neoplastic cells showed rhabdoid features or vacuolated cytoplasm. Immunohistochemically, the neoplastic cells were positive for vimentin and S100 and partly positive for neuron-specific enolase and glial fibrillary acidic protein but were negative for von Willebrand factor, desmin and α-smooth muscle actin. Ultrastructurally, the neoplastic cells had abundant cytoplasmic processes and desmosome-like structures. Cytoplasmic inclusions of rhabdoid-featured cells in HE sections were composed of aggregates of intermediate filaments, and cytoplasmic vacuoles were identified as an invagination of cytoplasm. Although malignant peripheral nerve sheath tumor was suggested according to these results, the present case was diagnosed as a soft tissue sarcoma with rhabdoid features due to a lack of identification of the basal lamina under electron microscopy.     

Peripheral Nerve Sheath Tumor in the Pelvic Limb of a Domestic Rabbit (Oryctolagus cuniculus)

A 3-year-old, 4.0 kg, female, spayed Silver Marten cross rabbit (Oryctolagus cuniculus) was presented for evaluation of a suspected recurring spindle cell sarcoma following primary excision by the referring veterinarian. Physical examination revealed a large subcutaneous mass on the left metatarsus and a fine needle aspirate revealed suspected neoplastic spindle cells. Hematologic evaluation was unremarkable and a computed tomography scan revealed invasion of the mass into skeletal muscles but showed no evidence of metastatic disease. An amputation was performed, after which the patient recovered uneventfully. However, the rabbit was found suddenly dead at time of discharge from unknown cause. Gross, histologic, and immunohistochemical evaluation was performed, which diagnosed a peripheral nerve sheath tumor. To the authors’ knowledge, this is the first detailed case report of a lagomorph peripheral nerve sheath tumor specifically confirmed with immunohistochemistry. Wide surgical excision is considered the treatment of choice and may be combined with radiation therapy. Further research is required to determine risk factors, the role of adjunctive therapy, as well as long-term prognoses for peripheral nerve sheath tumors in rabbits.     

Peripheral nerve sheath tumour of the equine maxillary sinus

This case report describes an 8‐year‐old Belgian Warmblood mare that was initially diagnosed and treated for a dental sinusitis. Despite removal of the diseased teeth and appropriate treatment, the sinusitis did not resolve. Over time, growth of a peripheral nerve sheath tumour of the left maxillary sinus became apparent, with recurrence after surgical removal. The clinical, radiographic, computed tomography, magnetic resonance imaging and pathological (gross and histological) findings of the first reported peripheral nerve sheath tumour of the equine paranasal region are described and illustrated.     

Paravertebral malignant peripheral nerve sheath tumour (MPNST) in a horse

A 10‐year‐old Lipizzaner gelding was presented with intermittent ataxia and hindlimb weakness. Ultrasonographic examination identified a mass cranial to the tuber sacrale. A provisional diagnosis of a soft tissue sarcoma was made based on a biopsy specimen. Owing to the extensive nature of the tumour and the associated poor prognosis, the horse was subjected to euthanasia on humane grounds. A post mortem examination revealed a locally infiltrative soft mass within the left lumbosacral epaxial musculature. Histologically, an infiltrative neoplasm predominantly composed of pleomorphic spindle or stellate‐shaped cells was identified. Neoplastic cells exhibited strong S‐100 protein and GFAP expression and variable vimentin, NSE, NGFR and myoglobin expression. They were uniformly negative for pan‐cytokeratin, melan A, laminin and desmin. The ultrastructural examination revealed pleomorphic cells with long cytoplasmic processes and an absence of melanosomes. Based on these results, a diagnosis of malignant peripheral nerve sheath tumour was made. This report contributes further information to assist in the diagnosis of these poorly defined neoplasms in animals, especially when they occur in uncommon locations.     

Glomus tumours in the skin and subcutis of three horses

Three horses presented with variably painful, nonulcerated masses of the head or neck that were diagnosed as glomus tumours. Grossly, they were fleshy, pink to tan masses ranging from 0.4 to 9 cm in diameter, involving either the deep dermis and subcutis or the subcutis and underlying skeletal muscle. Microscopically, neoplastic epithelioid cells were arranged in sheets, cords and packets within lobules. The neoplastic cells frequently abutted and formed nodular bulges into large endothelium-lined vascular spaces, especially around the tumour periphery. Large nerve branches were associated with each tumour. As determined by immunohistochemistry, the neoplastic cells consistently expressed α-smooth muscle actin and vimentin, and some cells in two of the cases expressed desmin. A laminin- or collagen IV-positive basement membrane was demonstrated around individual tumour cells or small groups of cells in all three cases. Morphological features and immunohistochemistry supported the diagnosis of glomus tumour, most consistent with the solid type in humans. Applying a classification system used in humans, two of these tumours met criteria of malignancy (glomangiosarcomas). One horse was euthanized due to complications associated with recurrence and treatment-related necrosis and secondary infection.     

Histochemical and immunohistochemical characterization of chordoma in ferrets

Chordomas of the tip of the tail in 6 ferrets were examined using histopathological, histochemical and immunohistochemical procedures. Histopathologically, round neoplastic cells containing numerous cytoplasmic vacuoles of varying sizes, categorized as “physaliphorous cells”, were observed in the amorphous eosinophilic or pale basophilic myxoid stroma. Physaliphorous cells were arranged in lobules and in a “chordoid” or “cobblestone” manner. The neoplasms were diagnosed as benign chordoma without local invasion and metastasis. Histochemically, the cytoplasm of small neoplastic cells was positive for periodic acid-Schiff stain and alcian blue (AB) pH 2.5 and pH 1.0 stains, but negative for hyaluronidase digestion-AB pH 2.5 stain. All neoplastic cells were strongly stained with colloidal ion, negative for high iron diamine AB pH 2.5 and toluidine blue pH 2.5 stains, and positive for Mayer’s mucicarmine stain. Immunohistochemistry using antibodies directed against low-molecular-weight cytokeratins (CK18, CK19 and CK20), vimentin and mucin core protein (MUC5AC) revealed that neoplastic cells had both epithelial and mesenchymal elements. The expression of low-molecular-weight cytokeratins suggests that neoplastic cells acquired the properties of glandular epithelial cells and produced epithelial mucus. Furthermore, the expression of cytokeratins, vimentin, S100 protein, brachyury and epithelial membrane antigen indicates that the neoplasms were equivalent to the classic type of human chordoma. Therefore, immunohistochemistry using these antibodies can be useful for the characterization of ferret chordoma.     

Thyroid follicular adenocarcinoma in a ferret

A 5-year-old male castrated ferret was presented to the Washington State University College of Veterinary Medicine for evaluation of progressive hair loss and a large, rapidly growing ventral neck mass. The patient had been diagnosed previously with an insulinoma, which was managed medically. Fine-needle aspirates of the neck mass were performed. The cytologic results were most consistent with epithelial neoplasia, likely a carcinoma; thyroid origin was considered likely based on tumor location and cell morphology. The tumor grew rapidly, and the owners elected euthanasia 1 week after examination. At necropsy, a circumscribed, ovoid mass disrupted the right cervical musculature next to the right lobe of the thyroid gland. Histopathologic evaluation revealed an infiltrative mass consisting of cuboidal cells arranged in solid sheets and irregular follicles enclosing colloid. The cells were large, with prominent nucleoli, and had a high mitotic rate. The histopathologic diagnosis was consistent with thyroid follicular adenocarcinoma. Immunochemical findings confirmed thyroglobulin production by neoplastic cells, but to a lesser extent than in normal ferret thyroid tissue. To our knowledge, this is the first case of thyroid follicular adenocarcinoma to be reported in a ferret, with only 1 other case of thyroid carcinoma, a C-cell carcinoma, described previously.     

Diagnosis of sciatic nerve tumour in two dogs by electromyography and magnetic resonance imaging

Sciatic nerve tumour was diagnosed in a Staffordshire Bull Terrier cross-bred and a Bichon Frise, both presenting with chronic left hind limb lameness. Neurological examination in each case was consistent with left sciatic nerve deficits and this was confirmed by EMG studies. Rectal examination of both dogs revealed a palpable intrapelvic mass that was not apparent on survey radiographs. A sciatic nerve tumour was identified using MRI in each case. Histological examination of tissue taken from the Staffordshire Bull Terrier cross-bred was consistent with a malignant peripheral nerve sheath tumour.