Differentiated embryonal rhabdomyosarcoma in a cow.

An embryonal rhabdomyosarcoma was found in the pleura of a 2-year-old Holstein cow after first delivery. The most predominant cells in the tumor were relatively small in size, but considerable numbers of more differentiated cells of larger sizes mingled with the small cells. The most differentiated cells were characterized by multinucleation, abundant cytoplasm containing cross-striated fibrils, intense immunoreactivity for desmin, and weak or negative reactivity for vimentin. Such cells, lacking mitotic activity and displaying weak or no reactivity for proliferating cell nuclear antigen, were considered to be malignant counterparts of myotubes or muscle fibers. This neoplasm seems to follow normal skeletal muscle embryogenesis, and to be capable of differentiation into the final stage of muscle development.     

Auricular rhabdomyosarcoma in a rat

An ear (auricular) neoplasm from a 1-year-old male rat was removed surgically and examined histologically. Macroscopically, the neoplasm was firm, white and measured (0.5 x 0.5 cm). Microscopically, the neoplasm was expansile, non-encapsulated, and composed of large, pleomorphic, polygonal to spindle-shaped cells containing multiple nuclei. Using immunohistochemical and chemical stains, the neoplastic cells were positive for vimentin, myoglobin, phosphotungstic acid haematoxylin and desmin, but had no immunoreactivity for cytokeratin or alpha-smooth muscle actin. On the basis of histopathological, immunohistochemical and histochemical stains, a diagnosis of auricular rhabdomyosarcoma was made. Although reported infrequently in human, this is, to the author's knowledge, the first report that describes the detailed gross, histopathological, histochemical and immunohistochemical findings of auricular rhabdomyosarcoma in a rat.     

What is your diagnosis? Perifemoral mass in a cow

Abstract: A 15‐year‐old female Simmental cross‐breed cow was presented to the Purdue University Veterinary Teaching Hospital for evaluation of a perifemoral soft tissue mass. Impression smears made from an excisional biopsy contained a population of pleomorphic mesenchymal cells with abundant, periodic acid–Schiff‐positive (PAS), intracytoplasmic granular material, and rare elongated multinucleated cells consistent with strap‐like cells. A second population of small round cells suggestive of lymphocytes or progenitor cells was also noted. A cytologic diagnosis of sarcoma was made, with rhabdomyosarcoma considered most likely based on the large amount of PAS‐positive material (presumed to be glycogen) and the rare strap‐like cells. Histopathologic sections contained an unencapsulated, densely cellular neoplasm composed of haphazardly arranged highly pleomorphic mesenchymal cells and a few small round cells. The mesenchymal cells were positive for vimentin, non‐specific muscle actin, and myoglobin, and negative for phosphotungstic acid‐hematoxylin, smooth muscle actin, and desmin. Glycogen granules were confirmed by transmission electron microscopy. A diagnosis of pleomorphic rhabdomyosarcoma was made. While cytologic findings may suggest rhabdomyosarcoma, cytologic features can be highly variable, and a definitive diagnosis usually requires cytochemical and immunohistochemical staining.     

Smooth Muscle Cells of the Bovine Cervical Stroma may have a Secretory, rather than a Contractile Function during Parturition

The bovine cervix contains a large amount of smooth muscle cells distributed over an outer muscular layer and within a stromal layer. The stromal layer exhibits no electromyographic (EMG) activity at parturition. This leads to the question whether the stromal smooth muscle cells of the bovine cervix are prepared to contract with parturition, or whether they have another function. To this end, cervical biopsies were repeatedly taken from 10 pregnant cows at day-185 and -275 of gestation, at spontaneous, uncomplicated calving and at 30 days after calving. The smooth muscle bundles of the stroma were immunohistochemically analysed (n = 5) with regard to their integrity and cellular density, and the degree of staining for connexin-43, smooth muscle actin α (SMA), desmin and vimentin. Additionally, the mRNA expression for connexin-43, SMA, desmin and vimentin was determined with RT-PCR (n = 5). The smooth muscle tissue was arranged in bundles, also at parturition. However, the cellular density of these bundles and the SMA mRNA expression were decreased at parturition. Additionally, the SMA staining and connexin-43 expression and staining remained constant during pregnancy and at parturition. This might indicate that stromal smooth muscle cells are not prepared to contract with parturition, in contrast to the myometrial smooth muscle cells. The smooth muscle cells, stained for SMA, also expressed vimentin, and the proportion of co-expression was increased at day-275 of pregnancy. This suggests that the stromal smooth muscle cells predominantly have a secretory function in cows.     

Rhabdomyosarcoma in a ferret (Mustela putorius furo)

A 5-year-old spayed male ferret showed a subcutaneous mass in the right lateral thoracic wall. Microscopic examination revealed that the neoplasm had proliferated in the subcutis with infiltration into the surrounding tissues. A packed bundle of large polymorphic neoplastic cells, containing abundant eosinophilic cytoplasm and a round to ovoid, occasionally bizarre nucleus, were arranged interwoven. The neoplasm had metastasized to the right axillary lymph node. The neoplastic cells were intensively positive for vimentin, desmin and myoglobin. Skeletal muscle type creatine phosphokinase-positive granules were detected in the cytoplasm. Ultrastructurally, various amounts of disorganized myofibrils with focal density resembling the Z-band were shown in the cytoplasm of the neoplastic cells. The neoplasia was diagnosed as rhabdomyosarcoma.     

Characterization of spindle cell component of ferret (Mustela putorius furo) adrenal cortical neoplasms – correlation to clinical parameters and prognosis

Adrenal cortical epithelial tumours are common in ferrets. A variant tumour type with prominent spindle cell proliferation has been identified. We characterized these variant tumours with light microscopy and immunohistochemical analysis and correlate these features to clinical parameters and prognosis. We classified 24 ferret adrenal cortical masses with recognizable spindle cell proliferation obtained from the AMC and AFIP databases, based on percentage of spindle cells present and features of malignancy. These masses were separated into hyperplastic nodules and adenomas (both with 1–24% spindle cells), ‘mixed’ adenomas (≥25% spindle cells), adenocarcinomas (1–24%) and ‘mixed’ adenocarcinomas (≥25% spindle cells). Tumours were evaluated immunohistochemically for smooth muscle actin (SMA) and estrogen receptor (ER) expression. Disease‐free interval (DFI) and survival time (ST) were calculated using Kaplan–Meier product limit method. Of 24 cases of spindle cell variant adrenal tumours, one was a hyperplastic nodule, 10 were adenomas, three were ‘mixed’ adenomas, six were adenocarcinomas and four were ‘mixed’ adenocarcinomas. The proliferative spindle cell cytoplasm was SMA‐positive (smooth muscle myocyte origin). ER positivity, seen in nine of 24 cases, was restricted to adenocarcinomas, ‘mixed’ adenomas and ‘mixed’ adenocarcinomas. DFI and ST were significantly reduced in ‘mixed’ adenocarcinomas or tumours with ER expression. DFI was significantly reduced in tumours with marked smooth muscle. The spindle cell component of these variant adrenal tumours is smooth muscle in origin. The presence of abundant smooth muscle, a more malignant histologic grade (‘mixed’ adenocarcinomas) and ER expression are significantly positively correlated to both decreased DFI and decreased ST.     

What is your diagnosis? Lingual mass in a dog

A 3.5-year-old female spayed Rat Terrier was presented for evaluation of a submucosal lingual mass. Fine-needle aspiration of the mass revealed a population of neoplastic cells composed predominantly of small, round cells and large, round to spindle-shaped cells, which occasionally had blunt cytoplasmic borders and multiple nuclei. The neoplastic cells had moderate to marked anisocytosis and anisokaryosis. The cytologic interpretation was malignant neoplasia, most likely sarcoma. Histopathologic examination of a biopsy specimen revealed an unencapsulated, poorly demarcated, moderately cellular neoplasm composed of individualized, infiltrative spindle cells. Elongate skeletal muscle cells with prominent, rectangular borders (strap cells) were observed. A morphologic diagnosis of rhabdomyosarcoma was made. Staining with phosphotungstic acid-hematoxylin revealed haphazardly arranged skeletal muscle cross-striations. Immunohistochemical staining results for vimentin, Myo D1, desmin, and myoglobin were positive, though staining intensity of tumor cells varied with the degree of differentiation. Using transmission electron microscopy, irregular, disorganized Z-bands were identified. Rhabdomyosarcomas are uncommon tumors in the dog, and rarely may involve the tongue or oral cavity. Cytologic evaluation of a rhabdomyosarcoma may reveal a pleomorphic population of cells and definitive diagnosis may require histologic examination, immunohistochemical staining, and electron microscopy.     

Establishment and characterization of canine rhabdomyosarcoma cell line CMS-C

A novel canine tumor cell line designated as the CMS-C cell line was established from pleomorphic rhabdomyosarcoma (RMS) raised in the prostate gland of a 14-year-old intact male mixed-breed dog. CMS-C cells displayed the same immunohistochemical characteristics (positive for vimentin and desmin and negative for cytokeratin and smooth muscle actin) as the original tumor cells and express myoD1 and UCP3, known as striated muscle-specific molecules, as shown by RT-PCR assay. Therefore, the established CMS-C cell line appears to be of rhabdomyoblast cell origin. The CMS-C cell line established from pleomorphic RMS will be a useful tool for further studies about canine RMS.     

Rhabdomyosarcoma in the abdominal cavity of a 12-month-old female donryu rat

Neoplasms of skeletal muscle origin are very rare in the rat. Recently, we experienced a case of rhabdomyosarcoma as a white mass involving the junction of the esophagus and stomach in the abdominal cavity of a 12-month-old female Donryu rat. Histopathologically, the neoplastic cells composing the mass invasively spreaded from the lamina propia to the tunica serosa in the stomach as well as the esophagus. Although the neoplastic cells varied in appearance, pleomorphic atypical cells with abundant eosinophilic cytoplasm were prominent. Some tumor cells were stained blue with phosphotungstic acid hematoxylin. The nuclei of spindle-shaped neoplastic cells were arranged longitudinally like beads. Multinucleate giant cells and mitotic figures were also frequently observed. Immunohistochemically, these neoplastic cells were positive for desmin and myoglobin, whereas they were negative for alpha-smooth muscle actin. Taken together these findings, this tumor was diagnosed as a pleomorphic rhabdomyosarcoma, probably derived from the muscle layer of the lower part of the esophagus. This is the first report of rhabdomyosarcoma in a Donryu rat.     

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.     

An immunohistochemical study of the distribution of intermediate filaments in the ovary of the emu (Dromaius novaehollandiae)

The immunohistochemical localization of the intermediate filaments desmin, vimentin and smooth muscle actin (SMA) in the ovary of the emu was described in the present study. The cortical region of the ovary contained developing and atretic primordial, pre-vitellogenic and vitellogenic follicles. Vimentin immunostaining was demonstrated in the granulosa cell layer of primordial, pre-vitellogenic and vitellogenic developing and atretic follicles. An interesting finding of the present study was the localization of SMA in fibroblasts located in the theca externa of late vitellogenic follicles. The presence of SMA in these fibroblasts suggests that they possess characteristics of smooth muscle cells.     

Differentiation of skeletal muscle Mesenchymal progenitor cells to myofibroblasts is reversible

Accumulation of intramuscular adipose tissue (IMAT) and development of fibrous tissues due to accumulation of collagen both affect meat quality such as tenderness, texture, and flavor. Thus, it is important for the production of high‐quality meat to regulate the amount of adipose and fibrous tissues in skeletal muscle. IMAT is comprised of adipocytes, while collagens included in fibrous tissues are mainly produced by activated fibroblasts. Both adipocytes and fibroblasts are differentiated from their common ancestors, called mesenchymal progenitor cells (MPC). We previously established rat MPC clone, 2G11 cells. As several reports implicated the plasticity of fibroblast differentiation, in the present study, using 2G11 cells, we asked whether myofibroblasts differentiated from MPC are capable of re‐gaining adipogenic potential in vitro. By treating with bFGF, their αSMA expression was reduced and adipogenic potential was restored partially. Furthermore, by lowering cell density together with bFGF treatment, 2G11 cell‐derived myofibroblasts lost αSMA expression and showed the highest adipogenic potential, and this was along with their morphological change from flattened‐ to spindle‐like shape, which is typically observed with MPC. These results indicated that MPC‐derived myofibroblasts could re‐acquire adipogenic potential, possibly mediated through returning to an undifferentiated MPC‐like state.     

Plasticity of endometrial epithelial and stromal cells—A new approach towards the pathogenesis of equine endometrosis

Equine endometrosis, a frequent cause of subfertility, is characterized by periglandular fibrosis, and no treatment exists. Endometrial biopsies not only contain diseased glands, but also contain healthy glands and stroma. Myoepithelial (ME) and myofibroblastic (MF) markers are calponin, smooth muscle actin (SMA), desmin and glial fibrillary acidic protein (GFAP). Epithelial vimentin expression indicates epithelial to mesenchymal transition (EMT). The aim of this immunohistochemical study was to investigate whether biopsies with endometrosis express MF and ME markers and vimentin. Compared to healthy areas, significantly higher percentages of endometrotic glands were lined by calponin‐ and vimentin‐positive epithelial cells, whereas periglandular fibrosis contained significantly higher percentages of stromal cells positive for vimentin, desmin and SMA and significantly less calponin‐positive stromal cells. The rare GFAP expression was restricted to endometrotic glands. Of these, the most frequent features of endometrotic glands were higher percentages of SMA‐ and vimentin‐positive stromal cells and the prominent epithelial calponin staining that occurred in 100%, 93% and 95% of examined biopsies. Results indicate plasticity of equine endometrial epithelial and stromal cells. Particularly, endometrotic glands show evidence for ME differentiation and EMT. The different expression of MF markers between stromal cells from healthy and endometrotic areas suggests functional differences. The characteristic changes in the expression of SMA, vimentin and calponin between endometrotic glands and healthy areas can be helpful to confirm early stages of endometrosis. The characterization of cellular differentiation may help to decipher the pathogenesis of endometrosis and could lead to therapeutic strategies.     

Rhabdomyosarcoma in a Silver Carp

Rhabdomyosarcoma is a primitive neoplasm that originates from skeletal muscle progenitor cells. In a routine inspection of a cyprinid farm in southwestern Iran, an approximately 2-year-old female Silver Carp was observed to have a raised mass located on the dorsolateral surface just caudal to the head. Macroscopic examination revealed a firm irregular fleshy pink mass (5 × 4 cm, depth = 3 cm) that appeared to arise from the subcutaneous musculature. Histologic sections were prepared using routine methods and separate sections were stained with hematoxylin-eosin and Massons’ trichrome. Microscopically, the tumor mass was composed of spindle cells that were densely packed and arranged in long interwoven bundles. The nuclei were vesicular and oval to elongated or cigar-shaped. Nuclear pleomorphism and multinucleate tumor giant cells were clearly evident. The neoplastic cell cytoplasm was eosinophilic with indistinct cell margins, and clear cross striations were observed in fibrils. The striated fibrils stained diffusely red with Masson’s trichrome. This account represents the first reported occurrence of rhabdomyosarcoma in Silver Carp Hypophthalmichthys molitrix.

Received July 10, 2015; accepted February 4, 2016     

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.     

Spontaneous deciduosarcoma in a domestic rabbit (Oryctolagus cuniculus)

Deciduosarcoma is a rare, hormonally dependent neoplasm with features of malignancy, previously reported only in rabbits enrolled in chronic toxicology studies involving estrogens with or without progestins. An exploratory laparotomy was performed on a 6-year-old pet Dutch dwarf rabbit following palpation of a 6-cm-diameter abdominal mass. Grossly, the mass was fleshy and nodular, adhered to but not appearing to originate from the small intestine, with a smaller mass of similar appearance involving the uterus, and an effaced mesenteric lymph node. Histologically, the mass was characterized by spindloid cells and large epithelioid cells with abundant pale eosinophilic vacuolated cytoplasm and an infiltrative pattern of growth. Giant cells with large, bizarre, hyperchromatic nuclei were common. Cells were positive by immunohistochemistry for vimentin and progesterone and estrogen receptors and negative for pancytokeratin (AE1/AE3), cytokeratin 18, desmin, alpha-smooth muscle actin (SMA), and CD10. Based on histologic and immunohistochemical findings, a diagnosis of deciduosarcoma was made.     

Casual discovery of a thoracic tumour showing histological features of undifferentiated pleomorphic sarcoma in a male Wistar laboratory rat

Sarcomas are neoplasms of mesenchymal origin, with a predominant cell population mimicking the organization of various soft tissues and/or bones. Previous categorizations also included the possibility of the presence of tissue macrophage-like (histiocytes) neoplasm cells, in a tumour described as malignant fibrous histiocytoma, but this group has been considered as a variety of undifferentiated pleomorphic sarcomas. Although this kind of malignancy is not rare in humans, only few cases have been reported in laboratory animals. We report an unusual single case of spontaneous tumour growth, detected by casual observation, in the left thoracic area of an 18-month-old male laboratory Wistar rat. Both this individual and his ancestors were not exposed to any known carcinogenic substance or radiation, thus suggesting the development of the neoplasm as a spontaneous event. The mass was extracted surgically under general anaesthesia, and slices were examined histologically and immunohistochemically, using photon microscopy. The pathologist reported the presence of a combination of fibroblasts and undifferentiated mesenchymal cells arranged in a storiform pattern. Immunohistochemistry was performed on the tissue using specific antibodies for several proliferation (Ki-67) and differentiation (S-100, CD-34, CD-68, pan-keratin, desmin and smooth muscle actin-SMA) markers. Positive reaction was observed for S-100, Ki-67, CD-68, desmin and SMA (limited) but not for CD-34 or cytokeratin.     

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.     

Myofibroblastic sarcoma of the pleura in a pig

An intrathoracic sarcoma from a 7-month-old female pig was studied by light and electron microscopy and immunohistochemistry. The tumor tissue of varied cell density consisted chiefly of spindle-shaped cells, some of which grew around blood vessels in a concentric fashion. Many tumor cells were positive for alpha smooth muscle actin and not for desmin, though some cells were reactive for both antigens. A majority of tumor cells had ultrastructural features characteristic of myofibroblasts, but a few cells resembled vascular smooth muscle cells in the synthetic state. This neoplasm may have arisen from vascular smooth muscle with features indicating a transition to myofibroblasts.