Follicular stem cell carcinoma: histologic,immunohistochemical, ultrastructural,and clinical characterization in 30 dogs

Diagnostic records of 30 primary and one metastatic follicular stem cell carcinomas in 30 dogs were reviewed. Neoplastic cells had a clear cytoplasm and formed lobules and nests surrounded by a basement membrane. Trichoepitheliomatous and apocrine differentiations were noted in 22 of 30 (73%) and 21 of 30 (70%) primary tumors, respectively. Glycogen was present in 20 of 20 (100%) tumors tested, suggesting tricholemmal differentiation. Antibodies against AE1/AE3 cytokeratin, vimentin, and melanA/MART1 stained 29 of 30 (97%), 29 of 30 (97%), and 12 of 27 (44%) primary tumors, respectively. Small amounts of melanin were identified in 14 primary tumors, either on the hematoxylin and eosin-stained section (n = 6), or on the Fontana-stained section (n = 8 of 14). Ultrastructural features of neoplastic cells included cell junction complexes, swollen mitochondria, neuroendocrine-like granules, and intracytoplasmic non-membrane-bound accumulation of proteinaceous material. Features of this neoplasm are consistent with a follicular stem cell origin. Follow-up information was available for eight dogs. Metastases developed in the draining lymph node at the time of excision of the primary tumor (n = 1) or subsequently (n = 3).     

Hepatobiliary neuroendocrine carcinoma in cats: a clinicopathologic, immunohistochemical, and ultrastructural study of 17 cases

Hepatobiliary neuroendocrine carcinoma was diagnosed in 17 cats in a period of 10 years. Seven tumors were of intrahepatic origin, one of which was a composite containing components of epithelial and neuroendocrine carcinoma. Nine tumors were of extrahepatic origin, and one tumor was located in the gall-bladder. The cats were adult and geriatric, and the male : female ratio varied according to tumor group. Hepatomegaly, anorexia, weight loss, and vomiting were the most common clinical signs observed in the cats with hepatic neuroendocrine carcinoma. The cats with extrahepatic neuroendocrine carcinoma showed these signs plus icterus (5/9) and high concentrations of hepatic enzymes. Histologically, the hepatic neuroendocrine carcinomas had two patterns, one with acinar structures separated by vascular stroma lined by cuboidal or columnar cells and the other solid with groups of anaplastic cells separated by vascular stroma. The composite tumor consisted of both bile duct carcinoma and neuroendocrine carcinoma. The extrahepatic neuroendocrine carcinomas and the gallbladder neuroendocrine carcinoma were characterized by solid sheets or groups of round to oval cells with vascular or fibrovascular stroma. Immunohistochemical examination of 10 of the neuroendocrine carcinomas revealed that all 10 stained with neuron-specific enolase; one bile duct carcinoma and the gallbladder carcinoma stained with chromogranin; four of five bile duct carcinomas and the gall bladder carcinoma stained with synaptophysin; and one bile duct carcinoma stained with gastrin. One cat with hepatic carcinoma had duodenal ulcer; in this cat, ultrastructural studies showed neurosecretory granules leading to the diagnosis of Zollinger-Ellison syndrome. In four cats in which necropsy was permitted, carcinomatosis (4/4), lymph nodes (4/4), lungs (2/4), and intestines (1/4) were the metastatic sites. Fourteen of the 17 cats were euthanatized during or immediately after surgery.     

Canine angiosarcoma: cytologic, histologic, and immunohistochemical correlations

BACKGROUND: Angiosarcomas (AS) are malignant tumors that arise from vascular endothelial cells and are common in dogs. Histologically, AS are markedly heterogeneous neoplasms that make interpretation by cytology difficult. OBJECTIVE: Our objective was to evaluate the cytologic features of canine AS and look for additional diagnostic criteria. METHODS: Cytologic specimens from 19 histologically and immunohistochemically confirmed cases of canine AS were extensively reviewed for cytologic features. We compared cytologic, histopathologic, and immunohistochemical findings. RESULTS: Neoplastic cells in 14 cytology specimens had a high-grade sarcomatous appearance, whereas in 4 specimens the cells were extremely pleomorphic, ranging from sarcomatous to epithelioid. In the remaining case, the neoplastic cells were low grade, spindle shaped, and monomorphic. Other relevant cytologic findings were blood contamination (18/19 cases), cellular cohesiveness (16/19), punctate cytoplasmic vacuolation (19/19), background neutrophilia (11/19) and eosinophilia (5/19), erythrophagocytosis (8/19), extramedullary hematopoiesis (8/19), and apoptotic leukocytes (14/19). Vasoformative features (ie, pseudoacinar structures) were observed in 7 of 19 samples. Histologically, 16 neoplasms had a proliferative pattern typical of well-differentiated canine AS. Three tumors were atypical poorly differentiated AS; 2 of these had a striking epithelioid pattern and 1 was a poorly differentiated spindle cell tumor with focal vascular differentiation. Immunohistochemically, tumor cells in 16 cases were positive for both endothelial markers tested (Factor VIII-related antigen [FVIII-ra] and CD31 antigen), 2 were positive for CD31 only, and 1 was positive for FVIII-ra only. The epithelioid AS were negative for cytokeratins. CONCLUSIONS: The cytologic characteristics of canine AS are widely heterogeneous, but supplementary findings can provide clues that are useful for making a cytologic diagnosis. Histologic and immunohistochemical confirmation is nonetheless warranted in all cases.     

Subcutaneous embryonal rhabdomyosarcoma in a dog: cytologic, immunocytochemical, histologic, and ultrastructural features

Abstract: A subcutaneous mass on the left antebrachium of an 11‐year‐old intact female English Pointer dog was evaluated presurgically by cytologic examination and immunocytochemical staining. The sample consisted of discrete, variably sized, markedly pleomorphic neoplastic cells that expressed vimentin with diffuse cytoplasmic staining, desmin with focal paranuclear staining, and myoglobin with diffuse cytoplasmic staining, consistent with a diagnosis of rhabdomyosarcoma. Lymphocytic and histiocytic markers were negative. Aspirates of the enlarged ipsilateral prescapular lymph node were positive for metastatic disease. Surgical excision of the tumor and lymph node were followed by histologic and electron microscopic examination. Histomorphologic appearance of neoplastic cells from the mass and the lymph node paralleled cytologic findings; the histologic diagnosis was round cell variant of embryonal rhabdomyosarcoma. By ultrastructural evaluation, cells contained numerous mitochondria and masses of cytoplasmic tangled myofilaments, features typical of rhabdomyoblasts. The dog received doxorubicin (30 mg/m²) every 3 weeks for 5 treatments. Local recurrence developed 6 months after resection but was not treated. Despite a guarded prognosis and untreated local recurrence, the dog was still alive 18 months after surgery. Cytologic evaluation and immunocytochemical staining were pivotal for the presurgical diagnosis of rhabdomyosarcoma.     

Cytologic, histologic, and immunohistochemical features of lingual liposarcoma in a dog

A 9-year-old female spayed mixed-breed dog was presented to the referring veterinarian with a history of decreased appetite and difficulty with prehension and swallowing because of a firm oval mass in the tongue. On cytologic evaluation of a fine-needle aspirate of the mass there were numerous round to polygonal cells organized individually or in loose clusters with rare branching capillaries. The cells had eosinophilic granular cytoplasm, round to oval nuclei, and occasionally indistinct borders. The cytologic diagnosis was granular cell tumor (GCT) of the tongue. Impression smears of a biopsy sample of the lingual mass contained similar eosinophilic granular cells with variable numbers of clear vacuoles in the background, numerous perivascular arrangements, and occasional lipoblasts, suggestive of liposarcoma. On histologic examination the tumor was composed of numerous lipocytes with rare foci of round eosinophilic granular cells without evidence of vacuolation; occasionally, atypical mitotic figures were seen. Immunohistochemically, the cells were uniformly negative for periodic acid-Schiff and did not express smooth muscle actin, desmin, or cytokeratin but were immunoreactive for vimentin and S100. A diagnosis of well-differentiated liposarcoma was made on the basis of morphologic and immunohistochemical results. Eosinophilic granular cells may be a component of well-differentiated liposarcoma and are not limited to GCT. Liposarcoma should be considered in the differential diagnoses of lingual tumors in the dog when cytological evaluation reveals eosinophilic granular cells consistent with GCT.     

Ehlers-Danlos-like syndrome in 2 dogs: clinical,histologic, and ultrastructural findings

Background: Ehlers-Danlos syndrome comprises a group of rare inherited connective tissue diseases characterized by skin hyperextensibility, joint laxity, skin and vessel fragility, and poor wound healing.
Objective: The purpose of this report was to describe the clinical, histologic, and ultrastructural findings in 2 dogs with collagenopathies consistent with Ehlers-Danlos syndrome.
Methods: Two dogs were examined clinically; skin extensibility index was calculated. Skin biopsies obtained from the dorsum were examined by light and electron microscopy.
Results: Both dogs had clinical signs of skin hyperextensibility and fragility, lower skin elasticity, vessel fragility, and poor wound healing. One dog had a hip dislocation, and the other had bilateral medial patellar luxation (grade II), subcutaneous hematomas produced by minimal trauma, and generalized periodontitis. Histologic and ultrastructural examination confirmed abnormalities in the structure and arrangement of collagen fibrils. Fibroblasts were characterized by variable dilatation of the rough endoplasmic reticulum, and anomalous elastic fibers (elaunin fibers) were present in the dermis.
Conclusion: Although the primary defects underlying collagenopathies in animals are still unknown, analysis of the ultrastructural changes in collagen fibrils and clinical findings could facilitate better characterization of these disorders in dogs.     

Pathogenesis of Staphylococcus aureus mastitis: bacteriologic, histologic, and ultrastructural pathologic findings

Three lactating cows were experimentally infected with Staphylococcus aureus ATCC 29740 (Newbould 305 strain). Cows were euthanatized 2 to 216 hours after inoculation. Bacteriologic and microscopic examinations showed that S aureus attached to epithelial cells of the mammary gland in vivo. The histopathologic changes observed were progressive swelling, vacuolar degeneration of epithelial layers, and multiple foci of epithelial erosions and ulcers throughout the ductal system. The cellular response of the infected glands was demonstrated by a rapid increase in the number of somatic cells in the secretion and by accumulation of neutrophils below, within, and on the epithelium of the teat and lactiferous sinuses. The inflammatory response did not prevent infection nor subsequent pathologic changes in the inoculated glands.     

Cytologic,histologic, and immunohistochemical features of maxillofacial alveolar rhabdomyosarcoma in a juvenile dog

Abstract: A 15‐month‐old castrated male dog with a history of intermittent epistaxis and sneezing was admitted for the examination of a maxillofacial mass. An impression smear of a biopsy sample from the cauliflower‐shaped gingival mass contained numerous round cells, 5–25 μm in diameter, which contained a moderate amount of clear to pale blue cytoplasm and resembled lymphoid cells. Mitotic figures were frequently observed. The mass was diagnosed as malignant round cell neoplasia. On histologic examination the tumor was composed of diffusely arranged, small, atypical round cells with a small amount of fibrovascular stroma. Immunohistochemically, the cells were negative for CD3, CD18, CD20, CD79α, cytokeratin, melan‐A, chromogranin A, α‐smooth muscle actin, and myoglobin but positive for vimentin and desmin. The cells also had strong positive nuclear staining for myogenin and MyoD1. A diagnosis of solid‐pattern alveolar rhabdomyosarcoma was made on the basis of morphologic and immunohistochemical results. Alveolar rhabdomyosarcoma should be considered in the differential diagnosis of tumors in juvenile dogs, especially when cytologic findings reveal round, undifferentiated cells.     

Malignant melanoma in a zebra finch (Taeniopygia guttata): cytologic, histologic, and ultrastructural characteristics

An approximately 3-year-old adult male zebra finch (Taeniopygia guttata) was diagnosed with malignant melanoma. The large darkly pigmented tumor was located in the coelom, extended from the apex of the heart to the cloaca, and was adherent to the intestines and the ventriculus. Dark small masses (likely metastases) were observed in the lungs. Cytologically, the neoplasm consisted mainly of round to oval cells with brown or pale blue to blue-brown pigment. Lesser numbers of cells were stellate to dendritic with abundant amounts of brown pigment granules or were markedly pleomorphic with variable amounts of pigment. Histologically, the tumor consisted of dense sheets and aggregates of infiltrative melanocytes that were negative for S-100 and Melan A. A few cells were consistent with "signet-ring" melanocytes. Melanocytes examined by electron microscopy contained typical structures, mainly premelanosomes and melanosomes, of this cell type. However, melanocytes with marked pleomorphism also contained intracytoplasmic aggregates of filaments, consistent with previously reported ultrastructural findings in signet-ring or rhabdoid melanoma of nonavian species.     

Characterization of uterine granular cell tumors in B6C3F1 mice: a histomorphologic, immunohistochemical, and ultrastructural study

The granular cell tumor is most often a benign neoplasm of uncertain origin. Four uterine granular cell tumors in control and treated female B6C3F1 mice were identified in chronic studies at the National Toxicology Program. Two tumors occurred in untreated control animals and 2 in treated animals receiving different compounds. Tissue sections were evaluated histologically and stained with hematoxylin and eosin, periodic acid-Schiff with diastase resistance, Masson's trichrome, toluidine blue, phosphotungstic acid-hematoxylin, and stained immunohistochemically with a panel of antibodies to muscle (desmin, alpha smooth muscle actin), neural (S-100, neuron specific enolase), epithelial (wide-spectrum cytokeratin), and macrophage (F4/80) markers. The main histomorphologic feature of tumor cells was the presence of abundant cytoplasmic eosinophilic granules that stained positive for periodic acid-Schiff with diastase resistance. Tumors varied in appearance and were comprised of sheets and nests of round to polygonal cells with distinct borders. Nuclei were hyperchromatic, pleomorphic, and centrally to eccentrically located and often contained single nucleoli. Occasional multinucleated giant cells were observed. Tumors were pale pink and homogeneous with trichrome stain and negative with toluidine blue. Three tumors had positive to weakly positive immunoreactivity for desmin, and 1 was positive for alpha smooth muscle actin. Expression of S-100, wide-spectrum cytokeratin, and neuron-specific enolase was negative for all tumors. Ultrastructurally, prominent electron-dense cytoplasmic granules were abundant and contained secondary lysosomes with heterogeneous lysosomal contents. The characteristics of these uterine granular cell tumors were suggestive of a myogenic origin.     

Granular cell tumor in the testis of a rabbit: cytologic, histologic, immunohistochemical, and electron microscopic characterization

An adult pet rabbit (Oryctolagus cuniculus) presented with a progressively enlarging left testicle. Because neoplasia was suspected, the testicle was removed. Pathologic evaluation (exfoliative cytology, histopathology, immunohistochemistry, electron microscopy) indicated that the tumor was an intratesticular granular cell tumor. Granular cell tumor has not been described in rabbits before and has not been described in the testes of animals or postnatal humans. Granular cell tumor should be included as a differential diagnosis of testicular tumors, particularly Leydig cell tumor, because of potential histologic similarities.     

Retrospective study of 338 canine oral melanomas with clinical, histologic, and immunohistochemical review of 129 cases

Diagnostic records from 338 canine oral melanomas in 338 dogs received at the Veterinary Medical Diagnostic Laboratory (1992-1999) were reviewed. Of these tumors, 122 plus an additional 7 metastatic melanomas of unknown origin were selected for clinical follow-up, histologic review, and immunohistochemistry. Chow Chow, Golden Retriever, and Pekingese/Poodle mix breeds were overrepresented, whereas Boxer and German Shepherd breeds were underrepresented. There was no gender predisposition and the average age at presentation was 11.4 years. Forty-nine dogs were euthanized due to recurrence or metastasis. The average postsurgical survival time was 173 days. The gingiva and the labial mucosa were the most common sites. Most tumors were composed of either polygonal cells (27 cases, 20.9%), spindle cells (44 cases, 34.1%), or a mixture of the two (polygonal and spindle) (54 cases, 41.9%). Clear cell (3 cases, 2.3%) and adenoid/papillary (1 case, 0.8%) patterns were uncommon. The metastases of 6/6 oral melanomas had morphologic and immunohistochemical features similar to those of the primary tumors. Immunohistochemically, Melan A was detected in 113/122 oral (92.6%) and 5/7 (71.9%) metastatic melanomas. Only 4/163 nonmelanocytic tumors were focally and weakly positive for Melan A. Antibodies against vimentin, S100 protein, and neuron-specific enolase stained 129 (100%), 98 (76%), and 115 (89.1%) of 129 melanomas, respectively. Antibodies against other melanocytic-associated antigens (tyrosinase, glycoprotein 100) did not yield adequate staining. We conclude that Melan A is a specific and sensitive marker for canine melanomas.     

Fibrosarcoma with typical features of postinjection sarcoma at site of microchip implant in a dog: histologic and immunohistochemical study

A 9-year-old, male French Bulldog was examined for a subcutaneous mass located at the site of a microchip implant. Cytologic examination of the mass was suggestive of a malignant mesenchymal neoplasm. Histologically, the mass was confirmed as a high-grade infiltrative fibrosarcoma, with multifocal necrosis and peripheral lymphoid aggregates. By immunohistochemistry, the sample was investigated for vimentin, smooth-muscle actin (SMA), CD3, CD79alpha, and CD18. All the neoplastic cells were positive for vimentin. Scattered cells at the periphery of the lesion were also positive for SMA, highlighting a myofibroblastic phenotype. The lymphoid cells were positive for CD18 and CD3. No aluminum deposits were detected by the aurintricarboxylic acid method. A diagnosis of fibrosarcoma morphologically similar to feline postinjection sarcomas was made. Fibrosarcomas at the site of injections have been reported in dogs and ferrets. Furthermore, neoplastic growth at the site of microchip implant in dog and laboratory rodents has been described.     

Morphological, immunohistochemical, and ultrastructural findings in an embryonal rhabdomyosarcoma of a newborn thoroughbred foal

This study reports the rare case of a rhabdomyosarcoma in the shoulder of a newborn Thoroughbred foal. Histomorphologic, immunohistochemical, and ultrastructural investigations revealed an embryonal rhabdomyosarcoma with various differences: small round undifferentiated cells, rhabdomyoblast-like neoplastic cells, and spindle-shaped cells. Desmin expression appears in 30% of small round cells and in as many as 90% of spindle-shaped cells. Vimentin co-expression occurs in 90% of small round and spindle-shaped neoplastic cells but only in few (30%) rhabdomyoblast-like cells. Myoglobin was detected in 50% of the rhabdomyoblast-like cells and in 30% of spindle-shaped cells but not in small round neoplastic cells. This is the first time that rhabdomyosarcoma markers Myf-4 and Myo D1 were found in horses. Myf-4 and Myo D1 were expressed in about half of the neoplastic cells in immediately fixated specimens. Ultrastructural sarcoma-like packets and formations of primitive Z-bands, as well as a discontinuous basal lamina, were found and served to confirm our diagnosis.

Introduction

Malignant neoplasia of striated muscles in domestic animals represents uncommon findings.[1] Only single cases of rhabdomyosarcoma in horses of varying ages (2-19 years) and races (Quarter horses, Appaloosas, Thoroughbreds, ponies) were described. [2, 3, 4, 5, 6 and 7]In human medicine, histopathologically different partly overlapping types of rhabdomyosarcoma are distinguished, reflecting stages of differentiation and the morphology of the neoplastic cells[8 and 9]: (1) embryonal rhabdomyosarcoma (subtypes: spindle cell, small cell) and its botryoid variant; (2) alveolar (synonymous with juvenile) rhabdomyosarcoma; and (3) pleomorphic (synonymous with adult) rhabdomyosarcoma.In veterinary pathology, rhabdomyoma, rhabdomyosarcoma, and “embryonal rhabdomyosarcoma of the urinary bladder in dogs” are actually distinguished histopathologically.[10] But in older references the nomenclature of human medicine is used.In horses, alveolar rhabdomyosarcoma occur in the extremities.[2 and 5] Botryoid rhabdomyosarcomas were described in the equine uterus [4] and the urinary bladder. [7] Pleomorphic rhabdomyosarcomas are found in the tongue [6] and inside the thorax. [3]Differential diagnoses include fibrosarcoma, melanoma, anaplastic lymphosarcoma, and anaplastic carcinoma. Immunohistochemical[10, 11 and 12] and ultrastructural [13 and 14] investigations are helpful in confirming a diagnosis of rhabdomyosarcoma.Myo D1, Myf-3, and Myf-4 belong to a group of myogenic regulatory proteins whose expression determines commitment and differentiation of primitive mesenchymal cells into skeletal muscle.[15] Myf-3 protein, the human homologue of the murine Myo D1 gene product, is a factor capable of myogenic determination, whereas Myf-4, the human homologue of myogenin, is required downstream as a differentiating factor. [15]This paper reports morphologic, immunohistochemical, and ultrastructural findings of an unusual case of embryonal rhabdomyosarcoma in the right shoulder of a newborn foal.

Case history

A newborn Thoroughbred foal showed a swelling in the right shoulder. This thickening was growing fast and at the age of 3 months a carneous mass of 5 kg was removed surgically by the local veterinarian. A distal dehiscence of the suture appeared and the swelling recurred within the following 3 months.

Clinical findings

When the foal was presented to the veterinary clinic at the age of 6 months, it showed a good general condition and no evidence of lameness. The right shoulder laterally showed a thick firm swelling of 20 cm. (Fig 1). At the caudodistal edge, a 10 cm deep fistula was found and sonographic examination showed multiple cysts within the tissue. Cytologic examination of the cyst fluid revealed numerous neutrophils and some adipocytes. Additional clinical findings revealed mild blood leukocytosis.