Electron microscopy of mast cells in the venous wall of canine liver

Although many mast cells locate under the endothelial layer along the sublobular veins in canine liver, the cell function remains to be fully defined. To establish the nature of the canine mast cell, the mast cells were examined by electron microscopy. A few monocytes contacted with luminal surface of endothelial cells under which mast cells situated. To confirm the chemotaxis of monocyte by hepatic mast cells, the hepatic venous vessels were treated with a histamine releaser (compound 48/80). The monocytes invaded into the subendothelial layer and extended their pseudopodium to the degranulated mast cells. It presumes that some mediators within the mast cell granules might act as a chemotactic substance to the monocyte. On the contrary, mast cells were migrating from subendothelial layer to venous lumen under normal condition. The migrating mast cell showed strong acid phosphatase reaction in their granules. It suggests that the granules of migrating mast cell became visible to acid phosphatase activity by a physical force such as contact stimulation, and that a part of mast cells remigrate from the venous wall to other places by the blood flow. Furthermore, hepatic mast cells were revealed to contain both endothelin-1 and histamine in their granules by immunocytochemistry. As these substances have an activity of stronger venous constriction, it seems that the mast cells play an important role in the blood flow regulation of the canine liver, mast cell, monocyte.     

Clinicopathological Features of Hemangioma in Two Young Horses

The clinicopathological and immunohistochemical features of two vascular tumors in two young horses are described in the present work. These animals were referred to the Veterinary Teaching Hospital of the University of Córdoba because of the presence of hyperpigmented plaques located in the medial aspect of the left leg and also around the hock (case 1) and in the right front leg (case 2). Some of the lesions showed deep ulceration and severe protrusion with abundant bleeding. The histopathological study revealed that lesions were composed of nonencapsulated, proliferated, closely packed small blood vessels, some of which showed irregular shape, whereas others were similar to capillaries, arterioles, and venules. Neoplastic cells expressed vimentin and factor VIIIar, suggesting their endothelial nature, whereas in the wall of some proliferated vessels, some cells expressed vimentin, α-smooth muscle actin and desmin, an immunophenotype consistent with pericytes, and small muscle cells. These features agree with those reported in human juvenile hemangioma rather than with hemangioma in adult horses.     

Neuromuscular and vascular hamartoma of the small intestine in an F344 rat

An intestinal mass was found in the border area of the jejunum and ileum of a 110-week-old male F344 rat. Histopathologically, the mass protruded into the lumen and was covered with intestinal epithelium, exhibiting a normal architecture. The lesion was located in the submucosa and consisted of loose connective tissue, smooth muscle, scattered ganglion cells, and blood vessels of various sizes. Although these components showed an irregular and disordered structure, no cellular atypia, increased proliferation activity, or invasive growth to adjacent tissues were detected. Immunohistochemical analyses revealed that smooth muscle, ganglion, and endothelial cells were positive for α-smooth muscle actin and vimentin, S-100, and CD34 and von Willebrand factor, respectively, indicating maturation of these cells. Thus, the mass was diagnosed as a neuromuscular and vascular hamartoma of the small intestine. To the best of our knowledge, this is the first report of this type of lesion in rodents.     

Conjunctiva-associated lymphoid tissue (CALT) in normal and Bordetella avium-infected turkeys

Conjunctiva-associated lymphoid tissue (CALT) was characterized in normal and Bordetella avium-infected turkey poults during the first 5 weeks of life. At 1, 5, 12, 19, 25, and 33 days post-hatching (DPH), upper and lower eyelids were examined by gross, histologic, and electron microscopic techniques. CALT was confined to the proximal part of the lower eyelid near the conjunctival fornix; it appeared by 5 DPH as individual lymphoid nodules and as dense masses by 19 DPH. In the upper eyelid, CALT was present only as isolated nodules. Histologically, CALT was composed of dense lymphocyte infiltrates within subepithelial connective tissue, intraepithelial lymphocytes, and flattened lymphoid-associated epithelium that lacked goblet cells. Germinal centers were in CALT by 19 DPH. By scanning electron microscopy, epithelial cells over lymphoid areas were flat and had short, irregular microvilli; non-lymphoid areas were covered by cells with tall, regular microvilli. Transmission electron microscopy revealed that with increasing age of birds, the epithelium over conjunctival lymphoid infiltrates became progressively flattened and infiltrated by lymphocytes. Some blood vessels in CALT had high endothelial cells; lymphocytes were in the lumen and between or beneath endothelial cells. In B. avium-infected poults, CALT was increased, developed earlier, and contained more germinal centers than in normal poults. We conclude that CALT of turkeys closely resembles other mucosal lymphoid tissues and may serve as a site for local antigen uptake.(ABSTRACT TRUNCATED AT 250 WORDS)     

Phenotypic characterization of normal and neoplastic canine endothelial cells by lectin histochemistry

Cell surface glycoconjugate expression of endothelial cells in canine cutaneous hemangiomas and hemangiosarcomas was compared to normal cutaneous endothelial cells using eight different lectins (with and without neuraminidase pretreatment) in an indirect immunoperoxidase technique. Direct comparison of lectin binding pattern of neoplastic endothelial cells with adjacent normal endothelial cells revealed minor changes in the binding intensity of several lectins (enhanced: Wheat germ agglutinin [WGA]; reduced: Griffonia simplicifolia-I [GS-I], Ricinus communis agglutinin-I [RCA-I], Soybean agglutinin after neuraminidase pretreatment [Neu-SBA], and Wheat germ agglutinin after neuraminidase treatment [Neu-WGA]). Neoplastic endothelial cells in some tumors exhibited varying binding of Ulex europaeus agglutinin-I (UEA-I; not binding to normal canine endothelial cells) and no Soybean agglutinin (SBA) binding (variably binding to normal endothelial cells in small cutaneous vessels). Lectin binding of neoplastic cells was rather heterogenous within one tumor compared to the uniform binding pattern of normal endothelial cells. These lectin binding studies demonstrate the phenotypic heterogeneity of neoplastic endothelial cells, indicating changes of cell surface glycosylation during neoplastic transformation.     

Corneal invasion by hemangiosarcoma in a horse

A 15-year-old gray Arabian gelding presented for evaluation of a lateral limbal mass extending across approximately 30% of the cornea. Grossly, the raised mass appeared nonpigmented, smooth, and irregular in shape, with an area of central necrosis and serosanguinous discharge. The mass was removed via lamellar keratectomy and histopathologic evaluation revealed features characteristic of hemangiosarcoma (HSA), including irregular vascular channels lined by a plump spindle cell population. Immunohistochemistry results showed that the neoplastic cells lining the vascular channels present diffuse and strong cytoplasmic reaction with von Willebrand Factor and the perivascular spindle cells exhibit moderate cytoplasmic reaction for smooth muscle actin. A lack of cytokeratin staining definitively excluded a diagnosis of atypical squamous cell carcinoma. Smooth muscle actin staining of the perivascular cells adjacent to the neoplastic endothelial cells is not a feature commonly described in HSA and has not been reported in previous cases of equine HSA. The horse remained in good health 21 months postkeratectomy and has exceeded the survival time of previously documented equine ocular HSA cases where more extreme surgical excision was performed.     

Hemangiosarcoma with widespread metastasis that originated on the metatarsal pad of a Java sparrow (Padda oryzivora)

A five-year-old male Java sparrow (Padda oryzivora) was examined because of the appearance of a purple spot approximately 1 mm in diameter on the right metatarsal pad. Seven months later, the spot spread to the entire right leg, and the bird died. At necropsy, multiple neoplastic masses on the right leg reached up to the pelvic cavity. There was one similar mass on the right kidney and multiple red spots on the liver. Histopathological examination revealed that neoplastic cells proliferated solidly or formed abundant irregular blood vessels. Watanabe silver staining revealed that neoplastic cells were surrounded by argentophil fibers. Immunohistochemically, the neoplastic cells were positive for anti-human von Willebrand factor antibodies. This is the first report of hemangiosarcoma in a finch.     

Labelling of rat endothelial cells with antibodies to vWF, RECA-1, PECAM-1, ICAM-1, OX-43 and ZO-1

Labelling with endothelium specific monoclonal antibodies, von Willebrand Factor (vWF), rat endothelial cell antigen-1 (RECA-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), OX-43 and zonula occludentes-1 (ZO-1), was investigated in cryostat sections of vessels from rats of different ages using a confocal microscope. The results showed that labelling of the vWF was positive in endothelial cells from adult, fetal and different ages of embryonic rat. Labelling with RECA-1 was weakly positive in adult rat aorta and lung endothelial cells but not in embryonic yolk sac endothelial cells. Labelling using PECAM-1, ICAM-1 and OX-43 was negative in both adult and embryonic endothelial cells. ZO-1 showed positive but very weak reactivity in embryonic yolk sac endothelial cells. The expression of vWF on vessels from adult and 19.5-day fetal tissues was strongly positive. However, the expression of vWF in embryonic endothelial cells was dependent on the gestational age. While the 11.5-day yolk sac vessels stained weakly, staining gradually increased in 13.5-, 15.5- and 17.5-day-old yolk sac vessels. The results suggest that vWF is a reliable endothelial cell marker in rat vascular endothelial cells, including both fetal and embryonic stages.     

Diffuse bilateral hemangiosarcoma of the uterus in a dog

A 15-year-old female mongrel dog showed abdominal swelling, marked hemorrhagic ascites and vulvar discharge, and ovariohysterectomy was performed. Grossly, the uterus was enlarged bilaterally without apparent mass formation. Histologically, the uterine muscular wall was composed of proliferated sinusoidal vessels. In some areas, irregular and small vessels proliferated markedly, while in others, pleomorphic and atypical tumor cells forming irregular vascular structures were predominant. From these findings, the case was diagnosed as diffuse bilateral hemangiosarcoma of the uterus that invaded to the ovary and broad ligament. The relationship between the tumor and angiomatosis was discussed.     

Haemorrhagic nephritis and enteritis of geese: pathomorphological investigations and proposed pathogenesis

Haemorrhagic nephritis and enteritis of geese as a new disease was first described in Hungary in 1969. The authors identified the causative agent of the outbreaks occurring in 1969 as a polyomavirus by PCR in 2001. In order to study the pathogenesis of the virus, one-day-old goslings were infected with tissue homogenate that tested positive for polyomavirus by PCR. Morphological, light and transmission electron microscopic (TEM) examinations have revealed that goose haemorrhagic polyomavirus replicates in the endothelial cells of the blood vessels and capillaries of diseased birds. Infection causes damage and necrosis of the endothelial cells. The virus was not observed in the parenchymal cells. Oedema and haemorrhages found throughout the body may be due to the dysfunction or functional deficiency of endothelial cells damaged by the virus.     

Morphologic features of lymph vessels in caprine hemal nodes

The distribution and characteristics of lymph vessels in caprine hemal nodes were studied after glutaraldehyde fixation and epoxy resin embedding. Histologically, the lymph vessels were characterized by thin walls and wide lumens containing inspissated lymph in which a few cells were suspended. The lymph vessels contrasted sharply with adjacent blood sinuses that were filled with elements of circulating blood. A circumferential lymph vessel in the cortex joined radial branches in the medulla that met at the hilum to drain through a large efferent lymph channel. Ultrastructurally, the lymph vessel wall comprised endothelial cells supported by a continuous basal lamina, collagen fibrils, and adventitial reticular cells. The cytoplasm of endothelial cells had fenestrations, plasmalemma-associated vesicles, vacuoles, and focal splits that enclosed large compartments. Many compartments contained erythrocytes and lymphocytes. Modifications of the endothelial cells signified their endowment with features that favored transendothelial transport. The distribution of lymph vessels and the finding of only efferent lymph channels are related to the roles of hemal nodes in blood storage by hemoconcentration and in immune defense mechanisms.     

A comparative ultrastructural study of the pecten oculi in adult,juvenile, and nestling yellow‐legged gulls,Larus michahellis (Naumann, 1840)

This study aimed at examining the histological structure of the pecten oculi in the adult yellow‐legged gull, Larus michahellis, and at two moments of postnatal development: during the posthatch (nestling) and juvenile periods. Particular attention was paid to differences in the diameter of vessels, the thickness of the basement membrane, and ultrastructural features of endothelial and pigmented stromal cells. Capillary endothelial cells displayed numerous microvillous‐like folds projecting from their internal and external surfaces. Intercellular spaces between capillaries were occupied by pigmented stromal cells. The ultrastructure of pecten oculi underwent noticeable changes during postnatal development. The examination of the capillaries in nestlings, juveniles, and adults revealed that the formation process of vessels and pigmented stromal cells did not complete itself in the posthaching phase. The prominent feature of endothelial cells of capillaries in nestlings was that the microvilli were longer than in juvenile and adult cells, and the capillary lumen was therefore reduced. In this sense, their pigmented stromal cells showed fewer melanosomes, lacked intercellular spaces, and cellular junctions could still be observed. These results provide evidence that the pecten oculi during the posthatching phase maintains immature morphological features consistent with a role of pigmented stromal cells in the blood‐retina barrier.     

Feline systemic reactive angioendotheliomatosis: eight cases and literature review

A rare, multisystemic intravascular proliferative disorder was identified postmortem in eight cats. The majority of these cats died or were euthanized following episodes of dyspnea, lethargy, and anorexia. Microscopic examination revealed occlusive, intraluminal proliferations of spindle cells within small vessels. The heart was consistently involved, and myocardial dysfunction was the probable cause of illness in all cats. Immunohistochemically, the majority of intravascular cells expressed von Willebrand factor, and a smaller number expressed smooth muscle actin, compatible with a dual population of endothelial cells and pericytes, suggesting a reactive rather than a neoplastic process. Four cases of a similar feline vascular disorder from the veterinary literature are reviewed. The histopathology resembles reactive angioendotheliomatosis in humans, a benign cutaneous intravascular endothelial and pericytic proliferative condition. However, in contrast, this feline disease is multisystemic and fatal. We propose the name "feline systemic reactive angioendotheliomatosis" for this unique, idiopathic disorder of domestic cats.     

Endothelial cells and angiogenesis in the horse in health and disease—A review

The cardiovascular system is the first functional organ in the embryo, and its blood vessels form a widespread conductive network within the organism. Blood vessels develop de novo, by the differentiation of endothelial progenitor cells (vasculogenesis) or by angiogenesis, which is the formation of new blood vessels from existing ones. This review presents an overview of the current knowledge on physiological and pathological angiogenesis in the horse including studies on equine endothelial cells. Principal study fields in equine angiogenesis research were identified: equine endothelial progenitor cells; equine endothelial cells and angiogenesis (heterogeneity, markers and assessment); endothelial regulatory molecules in equine angiogenesis; angiogenesis research in equine reproduction (ovary, uterus, placenta and conceptus, testis); angiogenesis research in pathological conditions (tumours, ocular pathologies, equine wound healing, musculoskeletal system and laminitis). The review also includes a table that summarizes in vitro studies on equine endothelial cells, either describing the isolation procedure or using previously isolated endothelial cells. A particular challenge of the review was that results published are fragmentary and sometimes even contradictory, raising more questions than they answer. In conclusion, angiogenesis is a major factor in several diseases frequently occurring in horses, but relatively few studies focus on angiogenesis in the horse. The challenge for the future is therefore to continue exploring new therapeutic angiogenesis strategies for horses to fill in the missing pieces of the puzzle.     

Epidermal growth factor and epidermal growth factor receptor immunoreactivity in the endothelial cells of the uterine artery and its branches during different stages of the estrous cycle in the pig

The uterine artery and its branches are the most important vessels that supply the uterus with blood, nutrients and active substances. Epidermal growth factor (EGF) and its receptor (EGFR) are expressed in many tissues, including reproductive organs, and is involved in angiogenesis, embryo implantation and development as well as in proliferation and differentiation of various cells. The aim of our study was to determine EGF and EGFR immunoexpression in the uterine artery and its branches during the estrous cycle in the pig. The experiment was performed on cryostat sections of the uterine artery and its branches stained immunohistochemically by ABC method. Light microscopic observations revealed the phase-related immunoreactivity of EGF and EGFR in the endothelial cells of the uterine artery and its branches. The highest intensity of EGF and EGFR immunoreaction in endothelial cells of the uterine artery was observed in the follicular phase. A significant decrease in the intensity of EGF and EGFR immunoreactivity was found in the middle luteal phase. Similar results of the immunostaining were found with regard to EGFR. In the endothelium of the uterine arterial branches, a significant increase in the intensity of EGF and EGFR-immunoreactivity was observed in the middle luteal phase. A decrease in the intensity of EGF immunostaining was observed in the late luteal phase. The phase-related expression of EGF and EGFR in the endothelium of the uterine artery and its branches suggest the modulatory effect of EGF and its receptor on the uterine artery and the region supplying these vessels.     

Model of chronic pancreatitis in the dog

A model of chronic pancreatitis was developed and characterized in the dog. Pancreatitis was produced by infusion of oleic acid through a cannula in the accessory pancreatic duct. Biochemical changes included early and marked increases in serum amylase and lipase activities which returned to base line within 3 weeks, at which time the dogs were clinically normal. In dogs euthanatized within 2 weeks, pathologic changes included massive necrosis and hemorrhage, cystic spaces filled with fluid, and abscesses. Histologic features revealed acute exudative pancreatitis with pancreatic atrophy and fibrosis. In dogs killed between weeks 3 to 12, changes included: marked atrophy with remaining acini surrounded by remnants consisting of collapsed stroma, blood vessels, and pancreatic ducts; marked coarse fibrosis with scattered inflammatory cells and occasional acinar tissue; and large irregular pseudocysts.     

Cutaneous haemangiosarcoma of the lower eyelid in an elderly white cat

A case of cutaneous haemangiosarcoma of the left lower eyelid in a 15-year-old white domestic shorthair cat is reported. A protuberant red mass occupying one-third of the lower eyelid margin length was present. Intermittent haemorrhage occurred from the mass surface. Surgical biopsy had revealed a locally invasive tumour composed of numerous irregular blood-filled spaces lined by a single layer of plump endothelial cells and separated by thin fibrous septa. Mitotic activity was rare and the appearance was consistent with a low-grade haemangiosarcoma. The mass continued to enlarge and referral was sought. Due to financial constraints and the owner's wish for a single procedure, enucleation with an axial pattern flap based on the superficial temporal artery was undertaken. Histopathology of the excised tissue confirmed the presence of a well-differentiated haemangiosarcoma. Tumour-free excisional margins were confirmed, the surgical area healed uneventfully, and there has been no recurrence during the subsequent 16 months.     

Cutaneous leiomyosarcoma with visceral metastases in a White Carneau pigeon and literature review

Cutaneous leiomyosarcomas are malignant mesenchymal tumors of smooth muscle origin and are reported occasionally in avian species. A 14-y-old male laboratory White Carneau pigeon (Columba livia) was presented for surgical excision of a cervical soft tissue mass. Ultrasonography with color flow Doppler imaging revealed multiple cavitations of mixed echogenicity within the mass and vascularization. Histologically, the dermis and subcutis were expanded by a densely cellular multinodular mass comprised of fusiform cells forming haphazardly arranged broad streams and short interwoven bundles, often surrounding blood vessels and variably sized cavitations. Neoplastic cells were strongly immunopositive for desmin and α–smooth muscle actin, and negative for pancytokeratin, S100, and von Willebrand factor. Based on histopathology and IHC findings, the cutaneous mass was diagnosed as leiomyosarcoma (LMS). The pigeon died 312 d post-operatively. Postmortem examination revealed masses infiltrating the left and right pulmonary airways and one hepatic nodule, but no regrowth at the surgical site. Histologic and IHC evaluation of the pulmonary and hepatic masses were consistent with LMS, representing metastatic foci from the primary cutaneous LMS. Our case highlights the malignant behavior and histomorphologic features of cutaneous LMS in an avian species.     

Multiple acquired extrahepatic portosystemic shunts secondary to veno-occlusive disease in a young German shepherd

An 11-month-old German shepherd presented with behavioral abnormalities. Histopathologic analysis of liver biopsies taken during exploratory laparotomy revealed veno-occlusive disease caused by fibrosis of the hepatic central veins leading to secondary portal hypertension, development of shunting vessels, and hepatic encephalopathy. The fibrosis was likely congenital in origin.     

Hematogenous metastasis of embryonal rhabdomyosarcoma originating from skeletal muscle in a young dog.

An 8-month-old, intact male Golden Retriever with a history of left forelimb lameness for 2 months was presented to the Veterinary Medical Teaching Hospital of Konkuk University (Seoul, Korea). Results of a physical examination revealed a mass in the left axillary region. A thoracic radiography showed an osteolytic lesion in the scapula and the presence of a soft tissue density from the thoracic wall to the scapula. A computerized tomography revealed a mass invading into the scapula, and small nodules in the lung that suggested metastasis. At necropsy, a pale-yellow, irregular, firm, 8 x 10 x 5 cm mass extended from axillary region and destroyed the scapular. In addition, small nodules were noted in the lung. On microscopic examination, the mass consisted of round-to-oval cells, with eccentrically located hyperchromatic nuclei and eosinophilic cytoplasm in fibromyxoid stroma. Tumor cells were observed in blood vessels in the primary mass. Tumor cells strongly expressed vimentin, desmin, and myoglobin. In phosphotungstic acid-hematoxylin staining, cross-striations were detected in rhabdomyoblasts. In periodic acid-Schiff reaction, only a few cells were detected. The diagnosis was primary rhabdomyosarcoma of the appendicular muscle of a young dog. The tumor presumably originated in the skeletal muscle of the limb, invaded into the adjacent scapular bone, and metastasized to the lung.