Mediastinal Schwannoma in a horse-cytologic,histologic, and immunochemical evaluation

A 31-kg mediastinal mass from a 24-year-old horse is described. Cytologic imprints contained numerous spindle cells with indistinct cytoplasm and multiple foci of palisading nuclei. Histologic sections had these same characteristics and areas of central fibrillar material between palisading nuclei (Verocay bodies). Within histologic sections, spindle cell cytoplasm had positive immunoreactivity for vimentin and S-100, whereas immunoreactivity was not detected for desmin and cytokeratin. Cytologic, histologic, and immunochemical evaluation confirmed a diagnosis of solitary schwannoma.     

Extranodal lymphoblastic lymphoma of suspected B-cell lineage in the gingiva of a racehorse,accompanied by mandibular osteolysis

A mass developed in the mandibular gingiva of a thoroughbred racehorse. When the horse could no longer eat unassisted, it was killed and immediately autopsied. Macroscopically, the mandible exhibited extensive osteolysis, with only a small amount of bone remaining around the tooth roots. The cut surface of the mass around the mandible consisted of neoplastic medullary tissue, in which osteogenesis was observed. The medullary tissue was composed of pleomorphic medium-sized to large cells, interlaced by collagen bundles. These cells had large, pale, round or ovoid, sometimes cleaved nuclei, with one or two prominent nucleoli. Mitoses were numerous. Electron microscopy showed that the cells in the medullary tissues were similar in shape to undifferentiated lymphocytes. Immunohistochemically, these cells were positive for B-cell associated antigen in the pre-B-cell stage. Our findings suggest that the horse had extranodal lymphoblastic lymphoma of suspected B-cell lineage, possibly originating from the lymphatic system of the gingiva. We consider that the osteolysis resulted from activation of osteoclasts by proliferation of the tumour cells.     

Vulvar leiomyosarcoma in a cat

A vulvar leiomyosarcoma was diagnosed in an 8-year-old mixed-breed, sexually intact cat. The cat had a history of vulvar swelling, bleeding and stranguria. The mass was located at the ventral commissura of the vulva. The cut surface of the mass was lobulated and greyish-white, with areas of necrosis and haemorrhage. Histologically, it was a spindle-cell sarcoma consisting of interlacing bundles of very elongated neoplastic smooth muscle cells and variably dense collagenous matrix separated by individual cells or streams. It also contained areas of focal necrosis and haemorrhage. The mitotic index ranged from seven to 12 mitoses per 10 high-power fields. Immunohistochemically, the spindle-shaped neoplastic cells were strongly positive for smooth muscle actin (SMA) and vimentin, whereas no immunoreactivity was obtained for desmin. The vascular elements of the internal control tissue also exhibited strong SMA and vimentin immunoreactivity. Based on the morphological and immunohistochemical features, the tumour was classified as a leiomyosarcoma.     

Increased concentrations of protein gene product 9.5 in the synovial fluid from horses with osteoarthritis

Our previous study established protein gene product 9.5 (PGP 9.5), a ubiquitin C-terminal hydrolase, as a specific cytochemical marker of synovial lining cells (type B synoviocytes) in the horse joint. The present study aimed to detect PGP 9.5 in the synovial fluid and shows that PGP 9.5 is a valuable marker of osteoarthritis in the horse. Immunohistochemical staining confirmed rich and consistent localization of PGP 9.5 immunoreactivity in the cytoplasm of synovial lining cells in the normal horse joint. Western blot analysis of synovial fluid from normal joints could detect a significant band corresponding to that contained in the brain and synovial membrane extracts. When 60 synovial fluid samples from normal and abnormal joints were assayed with an enzyme-linked immunosorbent assay (ELISA) system, the concentration of PGP 9.5 tended to be elevated in osteochondrosis dissecance, inflammatory arthropathy and intra-articular fracture, among which a statistically significant elevation was recognizable between the intra-articular fracture and the control. Thus, this study demonstrated the possibility that PGP 9.5, derived from synovial lining cells, may be a new biochemical marker for arthritic disorders of the horse.     

Dyspnea Caused by an Obstructive Tracheal Leiomyoma in a Horse: A Rare Case

Tracheal leiomyoma is a rare neoplasm of the respiratory tract in animals. This primary benign tumor arises from smooth muscle cells of the trachea. We report a 13-year-old female horse with clinical signs of severe airways obstruction. It had dyspnea, exercise intolerance, chronic progressive respiratory noises, and cyanosis in mucosa membranes. Radiography revealed a foreign body obliterating the luminal trachea. Endoscopic biopsy showed a 3 × 3-cm ovoid mass attached to the dorsal aspect of the trachea. Hematoxylin and eosin and Masson's trichrome stains showed dense spindle cells forming irregular bundles disposed in short interlacing fascicles. Nuclei were elongated with blunt ends, eosinophilic cytoplasm with indistinguishable cell borders, and low mitotic activity. This study is considered the first reported case of an obstructive intratracheal leiomyoma in the horse.     

Omental leiomyoma in a male adult horse

A well-circumscribed mass 70 x 35 x 28 cm in size and 41 kg in weight was detected at necropsy in a male adult horse within the omentum major without any association to the gastrointestinal tract. The tumor consisted of multiple white-to-yellow lobules and displayed a firm consistency. In addition, multiple cysts filled with blood-like fluid, and multifocal areas of necrosis were observed. Histologically, the mass consisted of slightly pleomorphic spindloid-shaped cells arranged in interlacing bundles containing elongated nuclei with blunt ends. The majority of tumor cells revealed a positive immunoreaction for alpha-smooth muscle actin, vimentin, and neuron-specific enolase and were negative for S-100, factor VIII-related antigen, and glial fibrillary acidic protein. Few tumor cells showed expression of desmin and c-kit. On the basis of macroscopy, histology, and immunohistochemistry, an omental leiomyoma was diagnosed.     

Zonal dermal separation: a distinctive histopathological lesion associated with hyperelastosis cutis in a Quarter Horse

This case report describes a distinctive deep cutaneous lesion in a 1-year-old Quarter Horse filly with hyperelastosis cutis. The horse had a typical clinical presentation of hyperelastic skin associated with a 6-month history of cutaneous wounds that developed following minor cutaneous trauma. Punch biopsies of skin from the affected horse were thinner than similar biopsies from an age- and breed-matched control. Significant microscopic lesions were not seen in cutaneous punch biopsies stained with haematoxylin and eosin and Masson's trichrome stains, but the ultrastructure of the dermis from the affected horse was characterized by variation in collagen fibre diameter and loose packing of collagen fibres within bundles. The horse was euthanized and necropsied, and full-thickness sections of skin were collected and examined microscopically. Affected skin was of normal thickness; however, the deep dermis contained a distinctive horizontal linear zone in which separation of collagen bundles resulted in the formation of large empty cleft-like spaces between the upper and lower regions of the deep dermis. We suggest the term 'zonal dermal separation' for this microscopic lesion. Incisional full-thickness skin biopsies should be taken in suspected cases of equine hyperelastosis cutis because punch biopsies may not obtain enough deep dermis to adequately represent pathological change in the skin of horses with this disorder.     

Spontaneous Rhabdomyosarcoma in a Common Marmoset ( Callithrix jacchus )

The common marmoset (Callithrix jacchus) is now widely used in various research fields, including toxicology. However, information about the background pathology of this species is scarce. Here, we report a case of rhabdomyosarcoma that spontaneously occurred in a common marmoset. A 44-month-old male common marmoset was euthanized due to bilateral hind limb paralysis. At necropsy, a 2×2×5-cm intramuscular mass was observed in the lower right back. Histologically, the mass was mainly composed of interlacing bundles of spindle-shaped tumor cells. Immunohistochemically, the tumor cells were positive for myogenin, desmin, vimentin and alpha-smooth muscle actin. Ultrastructurally, the tumor cells contained bundles of myofilaments with Z-band-like structures. Thus, the tumor was diagnosed as a rhabdomyosarcoma. To our knowledge, this is the first report of spontaneous rhabdomyosarcoma that was definitely diagnosed in the common marmoset.     

Immunohistochemical identification and localization of orexin A and orexin type 2 receptor in the horse gastrointestinal tract

The aim of the present study was to investigate the presence and the distribution of cells containing orexin A and orexin type 2 receptor in the horse stomach and gut, by means of immunohistochemical techniques.Orexin A was identified in the stomach fundic and pyloric regions and in the duodenum. In the same stomach regions, a large subset of orexin A-positive cells also showed orexin type 2 receptor-like immunoreactivity. Moreover, in the duodenum, many of them, seemed to store serotonin.Characteristically, enteric neurons or ganglia also displayed orexin A and, sometimes, orexin type 2 receptor immunoreaction.Orexin A and orexin type 2 receptor immunoreactivity was also found in the nerve fibers in the enteric submucosal layer.Our results, together with data present in the literature, could contribute to the understanding of complex mechanisms regulating the horse gut functionality that are depending very likely on the consequence of the co-operation of both a central and a peripheral control.     

Feline mammary sarcoma composed of cells resembling myofibroblasts

A recurrent mammary sarcoma from an 11-year-old, female domestic cat was studied by light and electron microscopy and immunohistochemistry. The tumor consisted of interlacing bands of spindle cells with elongated blunt-ended nuclei and variable amounts of stroma. Multinucleated tumor giant cells and mitoses were common. Ultrastructurally, tumor cells had abundant rough endoplasmic reticulum with dilated cisternae, a prominent Golgi complex, frequent mitochondria, bundles of intracytoplasmic filaments with focal densities, and discontinuous basal lamina-like material and cell junctions. These findings suggest that myofibroblast-like cells were the predominant type of tumor cell. Failure to demonstrate immunoreactivity for cytokeratins attested to the non-epithelial origin of these neoplastic cells. Uniform immunoreactivity with anti-vimentin antibodies and specific labelling of some tumor cells with antiserum to actin are compatible with an origin of this tumor from modified fibroblasts (i.e., myofibroblast-like cells). Tumors composed largely of myofibroblasts may be unique and warrant separate classification from other types of fibroblastic tumors in animals.     

Colocalization of gamma-aminobutyric acid immunoreactivity and acetylcholinesterase activity in nerve fibers of the mouse adrenal gland.

The present immunohistochemical and enzyme histochemical study showed gamma-aminobutyric acid (GABA) immunoreactivity and acetylcholinesterase (AChE) activity in the mouse adrenal gland. Weak GABA immunoreactivity was seen in clusters of chromaffin cells showing noradrenaline fluorescence. This finding suggests that both GABA and noradrenaline may be released from the granules of noradrenaline cells by adequate stimuli. GABA-immunoreactive varicose nerve fibers densely contacted adrenaline cells and large ganglion cells, but they were sparse in the periphery of clusters of noradrenaline cells. AChE activity was strong in a few large ganglion cells and weak in chromaffin cells showing noradrenaline fluorescence, and was found in numerous nerve bundles and fibers of the medulla. AChE-active nerve fibers more densely contacted noradrenaline cells than adrenaline cells. By using double labeling technique, numerous GABA-immunoreactive nerve fibers in the medulla were reactive for AChE in the same sections. These results suggest that both GABA and acetylcholine may be colocalized in the intra-adrenal nerve fibers and may have some secretory effects on the chromaffin cells.     

The distribution of nerve fibres immunoreactive for vasoactive intestinal peptide, calcitonin gene-related peptide, substance P and dopamine beta-hydroxylase in the normal equine larynx

The autonomic innervation of the mammalian respiratory system is complex, and involves a wide variety of peptide and non-peptide neurotransmitters which will have an important role in normal laryngeal function and the response to disease. This innervation has been partially described in the horse airway and lung, but there is no information on the equine larynx. This paper describes the expression and distribution of nerve fibres immunoreactive for vasoactive intestinal peptide (VIP), calcitonin gene-related peptide (CGRP), substance P (SP) and the adrenergic enzymatic marker dopamine beta-hydroxylase (DBetaH) in the mucosa of the equine larynx. The overall relative density of nerve fibres immunoreactive for the different antigens was VIP>CGRP>SP>DBetaH. There were differences in the distribution of nerve fibre types, although each antigen was found in nerve fibres adjacent to blood vessels and mucous glands. VIP -like immunoreactivity (VIP -Li) was particularly extensive in association with mucous glands. SP - and CGRP -like immunoreactivity (SP -Li, CGRP -Li) were also seen close to the epithelium, with occasional nerve fibres coursing beneath and between the epithelial cells. Fragments of SP -Li and CGRP -Li fibres were also present in large nerve fibre bundles and ganglionic cell clusters, but not in the neurons themselves. The density of nerve fibres immunoreactive for DBetaH was very low and restricted to blood vessels and mucous glands. There was marked variation in the density of nerve fibres at the different sites, with the greatest density, particularly for VIP, over the arytenoid cartilage. Immunoreactive nerve fibres were less plentiful over the epiglottis, and the density of all types of nerve fibres was low over the cricoid cartilage. Overall VIP -Li nerve fibres were the most plentiful.     

Vulvar lipoleiomyoma in a dog.

A vulvar neoplasm from a 5-year-old female Siberian Husky dog was removed surgically and examined histologically. Macroscopically, the neoplasm was firm, white, and measured 6 x 4 x 3 cm. Microscopically, the neoplasm was expansile, nonencapsulated, and composed of lobules of mature adipocytes ad-mixed with streams and bundles of well-differentiated smooth muscle cells. Immunohistochemically, the neoplastic cells had strong diffuse cytoplasmic immunoreactivity for alpha-smooth muscle actin and desmin, and no immunoreactivity for cytokeratin or vimentin. On the basis of gross, histopathologic, and immunohistochemical findings, a diagnosis of lipoleiomyoma was made. This is, to the author's knowledge, the first report of canine vulvar lipoleiomyoma.     

An immunohistochemical study of interstitial cells of Cajal (ICC) in the equine gastrointestinal tract.

The interstitial cells of Cajal (ICC) are c-kit immunoreactive cells of the gastrointestinal tract which are suggested to have a role in the control of intestinal motility. Cells with c-kit immunoreactivity have not been previously described in the gastrointestinal tract of the horse. Immunoreactivity for c-kit was revealed using immunohistochemical labelling with an anti-c-kit polyclonal antibody. Sections of normal gastrointestinal tissue were examined from 13 anatomically defined sites from stomach to small colon taken from horses free from gastrointestinal disease. Three types of c-kit immunoreactive cells were identified: spindle-shaped cells in the region of the myenteric plexus, stellate or bipolar cells in the circular muscle layer, and round cells in the submucosa. The round cells were shown to be mast cells with the use of toluidine blue staining, whereas the other c-kit immunoreactive cells did not exhibit metachromasia and were classified as ICC. This study will serve as a basis for future pathological studies in the horse.     

Influence of a high-protein diet on energy balance in obese cats allowed ad libitum access to food

The influence of a high-protein [HP, 47% of metabolizable energy (ME)] diet on energy balance was evaluated in obese cats allowed ad libitum access to food. Energy intake, body weight, body composition, energy expenditure, and concentrations of hormones and metabolites associated with carbohydrate and lipid metabolism (glucose, insulin, free fatty acids, triglycerides and leptin) were measured in cats after consuming either a moderate protein (MP, 27% of ME) or HP diet for 4 months. Indirect respiration calorimetry showed that resting and total energy expenditure (kJ/day) adjusted for either body weight or lean body mass was increased in cats consuming the HP in relation to MP diets. However, voluntary energy intake also was increased in the HP treatment and, thus, there was no difference in body weight between animals consuming the two diets. Body composition measurements using deuterium oxide dilution showed that dietary protein content did not alter amounts of either lean body mass or fat mass. No significant differences (p > 0.05) were observed between the two treatment groups for blood glucose, free fatty acid or leptin concentrations, although there was a trend (p = 0.054) towards an increase of serum insulin concentrations in the cats eating the HP diet. This study showed that short-term ad libitum feeding of an HP diet did not reduce food intake or promote weight loss in obese cats. However, energy expenditure was increased in the HP diet group and it is possible that this effect of HP might help promote weight loss when energy intake is restricted.     

Helix pomatia a hemagglutinin, a surface marker for bovine T-lymphocytes

Bovine peripheral blood lymphocytes (PBL) were isolated from blood collected from 6 cattle. After treatment with neuraminidase, 40 or 60% of the cells were shown to combine with Helix Pomatia A hemagglutinin (HP) depending whether a direct or indirect fluorescence technique was used. About 20% of the cells were Ig-bearing. With double staining fluorescence technique, it was shown that cells attaching to HP were not Ig-bearing and the reverse. With the aid of HP, covalently bound to Sepharose, Ig-bearing cells could be separated from cell populations attaching to HP. The fraction of cells forming rosettes with sheep erythrocytes was proportional to that of HP attaching cells both before and after fractionation on the HP column. It is therefore concluded that HP is a marker for bovine T-cells, and that this lectin may be used to separate B-cells from T-cells.     

Tail-base mass from a "horse of a different color"

A 14-year-old bay Thoroughbred gelding was presented for evaluation of a mass at the base of the tail. The mass had been present for 1 year, and recently had begun to increase in size. Additional masses were found around the eye and shoulder. A fine-needle aspirate of the tail-base mass revealed highly anaplastic round to polyhedral cells containing dark green to black cytoplasmic granules interpreted to be melanin. Histologically, the mass was composed of pleomorphic, poorly pigmented, round to polyhedral cells interpreted to be neoplastic melanocytes. With immunohistochemistry, the cells were positive for vimentin and S-100, but negative for pancytokeratin and Melan-A. The cytologic and histopathologic diagnoses were amelanotic melanoma. The horse was treated with cimetidine, but the tumor continued to progress. In this report, we describe the cytopathologic features of an aggressive amelanotic melanoma in a non-grey horse and emphasize the unique correlation between cytologic and histologic findings.     

Influence of dietary protein level on body composition and energy expenditure in calorically restricted overweight cats

High‐protein (HP) diets help prevent loss of lean mass in calorie‐restricted (CR) cats. However, it is not entirely known whether these diets also induce changes of energy expenditure during periods of CR. To investigate this issue, sixteen overweight cats were fed either a high‐protein [(HP), 54.2% of metabolizable energy (ME)] or a moderate‐protein [(MP), 31.5% of ME] diet at 70% of their maintenance energy intakes for 8 weeks, and energy expenditure, energy intake, body weight and composition, and serum metabolites and hormones were measured. While both groups of cats lost weight at a similar rate, only cats eating the HP diet maintained lean mass during weight loss. Indirect respiration calorimetry measurements revealed that both total and resting energy expenditure (kcal/d) significantly decreased during weight loss for both treatment groups. However, only cats eating the MP diet exhibited significant decreases of total and resting energy expenditures after energy expenditure was normalized for body weight or lean mass. Results from this study suggest that in addition to sparing the loss of lean mass, feeding HP diets to overweight cats in restricted amounts may be beneficial for preventing or minimizing decreases of mass‐adjusted energy expenditure during weight loss.     

Piloleiomyosarcoma in seven ferrets.

In each of seven ferrets (Mustela putorius furo) with leiomyosarcoma, a single dermal mass was identified and biopsied. Each mass consisted of a well-demarcated but nonencapsulated proliferation of large spindle- to strap-shaped cells arranged in interwoven bundles. The cells resembled the smooth muscle cells of the adjacent arrector pili muscles, but with marked nuclear pleomorphism. Immunohistochemical staining for smooth muscle actin, desmin, and vimentin was positive and staining for myoglobin and cytokeratin was negative. Follow-up on three of the ferrets indicates that the prognosis is good following complete surgical excision.