Automated tru-cut imaging-guided core needle biopsy of canine orbital neoplasia. A prospective feasibility study.

The purpose of this study was to evaluate the diagnostic value of imaging-guided core needle biopsy for canine orbital mass diagnosis. A second excisional biopsy obtained during surgery or necropsy was used as the reference standard. A prospective feasibility study was conducted in 23 canine orbital masses at a single centre. A complete ophthalmic examination was always followed by orbital ultrasound and computed tomography (CT) examination of the head. All masses were sampled with the patient still on the CT table using ultrasound (US) guided automatic tru-cut device. The most suitable sampling approach to the orbit was chosen each time based on the CT image analysis. One of the following different approaches was used: trans-orbital, trans-conjunctival or trans-masseteric. In all cases, the imaging-guided biopsy provided a sufficient amount of tissue for the histopathological diagnosis, which concurred with the biopsies obtained using the excisional technique. CT examination was essential for morphological diagnosis and provided detailed topographic information that allowed us to choose the safest orbital approach for the biopsy. US guided automatic tru-cut biopsy based on CT images, performed with patient still on the CT table, resulted in a minimally invasive, relatively easy, and accurate diagnostic procedure in dogs with orbital masses.

Sialic acid and sialyltransferase activity in serum and tissues of dogs with mammary tumors.

In humans, the glycosylation pattern of serum and of membrane glycoproteins is associated with invasiveness of tumors: specifically, α2,6-sialylation and α2,3-sialylation are associated with metastasizing and nonmetastasizing tumors, respectively. In turn, the type of sialylation depends on the activity of α2,6 or α2,3 sialyltransferase (ST) enzymes. Because of the high prevalence of metastasizing tumors with biological behavior similar to the human counterpart, female dogs with metastasizing neoplasms could provide a good animal model for investigating the potential roles of sialic acid (Sia) and ST enzymes in the pathogenesis of metastatic tumors. The aims of this study were (1) to validate a solid-phase method based on lectin staining of serum and tissue homogenates to investigate sialylation and ST activity and (2) to compare the results obtained with this method and with lectin staining and to collect preliminary information on sialylation and ST activity in dogs with (n = 8) and without (n = 8) mammary tumors. The data recorded in healthy dogs revealed that serum and tissue glycoproteins are prevalently characterized by a α2,6 sialylation, but ST-α2,3 seems to be the most active enzyme in both samples. Sia-α2,3 and ST-α2,3 activity decreases in serum and tissues of dogs with tumors, especially in a dog with metastasis, suggesting that the equilibrium between ST-α2,6 and ST-α2,3 activity shifts toward the former, as reported in humans.

A case of multiple unilateral corneal epithelial inclusion cysts in a dog.

A three-year-old neutered female Labrador cross was presented for investigation of two corneal epithelial inclusion cysts affecting the left eye. The aetiopathogenesis of the cyst formation was suspected to be traumatic. The cysts were removed successfully by superficial keratectomy, followed by a third eyelid flap. Histologically, both lesions were represented by cystic formations lined with multi-layered squamous epithelium, consistent with stromal inclusion of surface corneal epithelium. Complete recovery was achieved, and there was no recurrence at six month follow-up.

Classical and spermatocytic seminoma in the dog: histochemical and immunohistochemical findings.

In the light of earlier human studies, 43 canine tumours diagnosed as seminoma were examined histologically with haematoxylin and eosin and periodic acid-Schiff (PAS) stains, and immunohistochemically with a monoclonal antibody against human placental alkaline phosphatase (PLAP). Twenty tumours were positive for both PAS and PLAP and were therefore diagnosed as classical seminoma (SE). The other 23 tumours were negative for both PAS and PLAP and were therefore diagnosed as spermatocytic seminoma (SS). Tubules with carcinoma in situ (CIS) were present in the testicular parenchyma surrounding 15 SEs and nine SSs.

The distribution of oestrogen receptors in normal, hyperplastic and neoplastic canine prostate, as demonstrated immunohistochemically.

The role of oestrogens and their receptors (ERs) in prostatic growth and differentiation and in the progression of prostatic carcinoma has been well investigated in human medicine. In dogs, however, available reports on the expression of ERs in normal, hyperplastic or neoplastic prostates are few and controversial, or completely lacking. Three normal, three hyperplastic and nine neoplastic canine prostates were examined histologically, and immunohistochemically with a polyclonal antibody directed against human ERs. Evaluation of the percentage of immunolabelled cells was performed by digital image analysis. The study showed that ERs were expressed in the nuclei of epithelial and stromal cells in normal, hyperplastic and neoplastic prostates. All nuclei in normal and hyperplastic prostates were strongly and homogeneously immunolabelled by the antibody used. However, the percentages of immunolabelled nuclei in the nine prostatic carcinomas were greatly reduced, ranging from 36.34% to 66.73%. The severe loss of ER expression in canine prostatic carcinomas may account for the relative lack of differentiation of these tumours in the dog.

Multiple endocrine neoplasia type-I-like syndrome in two cats.

Multiple endocrine neoplasia (MEN) embodies a group of diseases in human patients and domestic animals that are characterized by hyperplasia or neoplasia, or both, of two or more endocrine tissues. The MEN-1 syndrome is associated with menin gene mutations that induce various combinations of parathyroid, pituitary, and pancreatic endocrine tumors in humans. Two male, Domestic Shorthair cats developed symmetric alopecia, insulin-resistant diabetes mellitus, and pituitary-dependent hyperadrenocorticism at 12 and 13 years of age. Examination of skin biopsy specimens revealed atrophic dermatosis associated with hyperadrenocorticism. In one cat, cutaneous lesions consistent with paraneoplastic alopecia associated with pancreatic adenocarcinoma also were evident. Multiple invasive pancreatic beta cell carcinomas, pituitary corticotroph adenomas, and thyroid C-cell and parathyroid chief cell hyperplasia were diagnosed on the basis of results of gross, histologic, and immunohistochemical findings in both cats. Pancreatic exocrine adenocarcinoma was diagnosed in both cats. One cat also had hepatocellular carcinoma. Exons 1-8 of the feline menin gene were sequenced and were found to bear 93% homology with the human gene sequence, and the corresponding amino acid sequences shared 98% homology. Purification of total RNA and amplification of cDNA from lesional tissues to document mutations in the feline menin gene sequence were unsuccessful. The combination of lesions observed was consistent with the diagnosis of MEN-1-like syndrome in both cats.

Gastric mucinous adenocarcinoma with cutaneous metastases in a dog: diagnosis by fine-needle aspiration cytology.

A 12-year-old, intact, male mixed-breed dog was presented with anorexia, vomiting and multiple cutaneous nodules on its neck, trunk and hindlimbs. Fine-needle aspiration cytology of the nodules was characterised by a pleomorphic population of cells arranged singly or in small cohesive clusters, embedded in an amorphous mucinous material stained positive by periodic acid-Schiff (PAS). Acinar structures were occasionally found. Cells appeared either small with scant basophilic cytoplasm or large with a histiocytic appearance. Large cells had cytoplasm filled with a PAS-positive granular material. A presumptive diagnosis of cutaneous metastases of a mucinous adenocarcinoma was made. A primary, gastric, signet-ring mucinous adenocarcinoma was confirmed at postmortem examination and by histopathology. To the authors' knowledge, this is the first report of a gastric mucinous adenocarcinoma with cutaneous disseminated metastases in a dog.

Immunohistochemical identification of Encephalitozoon cuniculi in phacoclastic uveitis in four rabbits.

PURPOSE: Encephalitozoon cuniculi is a microsporidium with a wide range of mammalian hosts. In rabbits it can be responsible for cataract and lens-induced uveitis (LIU). The aim of this study was to provide specific immunohistochemical demonstration and localization of E. cuniculi within the eye, in rabbits with LIU. MATERIAL AND METHODS: Four rabbits were presented with a white mass in the eye and iris discoloration. Complete ophthalmic examinations were performed and a presumptive diagnosis of LIU was made in all cases. Initial therapy with a topical steroid, atropine and systemic enrofloxacin was instituted while serologic (IFA or ICA tests) and cytologic lab results were pending. The final outcome in all cases was enucleation. Routine histology and immunohistochemistry (ABC method) with an antiserum anti-Encephalitozoon cuniculi were performed. RESULTS: Indirect immunofluorescence performed on one rabbit serum expressed a titer of 1 : 32; carbon immunoassay on the serum of the other three rabbits expressed a titer of 1 : 5120 in one, and a titer of 1 : 2560 in the other two cases. Histologically, an intraocular, locally extensive pyogranulomatous infiltration that partially filled the posterior chamber, encasing a wide anterior lens capsule break, was detected in all cases. Immunohistochemically, spores reacting with anti-Encephalitozoon cuniculi antiserum were present in all specimens, occasionally within macrophages and lens epithelial cells. CONCLUSION: Detection of E. cuniculi in rabbits with phacoclastic uveitis has been investigated in the past with different methods. Based on our results, we suggest that immunohistochemistry should be regarded as a useful tool both for specific demonstration of E. cuniculi and for its localization within tissues.

Immunohistochemical characterization of the leucocytic infiltrate associated with canine seminomas.

Both canine and human seminomas are typically associated with leucocytic infiltration, the role of which remains poorly understood. In this study, leucocytes infiltrating 10 canine seminomas were characterized. Monoclonal antibodies directed against CD18, CD11a, CD11b, CD11c, CD21, CD3, CD4, CD8 and Major Histocompatibility Complex class I and II (MHC I and MHC II) were also employed. Infiltrating leucocytes were located around vessels, adjacent to the thin fibrous septa between neoplastic lobules, and were also scattered singly amongst neoplastic cells. The more profuse infiltrates often had the appearance of follicular aggregates. Immunohistochemically, all the samples showed generally similar results. Most of the infiltrating cells were positive for CD18 and CD11a. Infiltrating cells were mainly T lymphocytes (CD3+), particularly of the CD8+ subset. B lymphocytes (CD21+) were detectable in almost all infiltrates; in the follicular aggregates they were centrally located, whereas T lymphocytes (CD3+) lined the periphery. CD11c+ cells were always more numerous than CD11b+ cells, demonstrating that if macrophages and antigen-presenting cells (APCs) were well represented, monocytes and granulocytes were practically absent. Almost all of the infiltrating cells were positive for both MHC I and MHC II antigens and, in nine samples, a large number of neoplastic cells expressing MHC I were detected. A few neoplastic cells expressing MHC II were observed in seven cases. The presence of CD8+ lymphocytes, together with the large number of both infiltrating and neoplastic cells expressing MHC I, suggests that the lymphocytes mediate a cytotoxic reaction against the neoplastic cells. This hypothesis may underlie the favourable prognosis frequently associated with canine seminomas.

Pathologic and immunohistochemical findings in a feline aortic body tumor.

The presence of a heart-base tumor was diagnosed by ultrasound imaging in a 10-year-old, female, domestic shorthaired cat presenting with dyspnea and pleural effusion because of the presence of a modified transudate. Hematology and clinical chemistry were unremarkable. The owner elected euthanasia. At necropsy, a locally extensive, firm, multilobulated nodule surrounded the pulmonary vein. The tumor was composed of lobules of large polygonal cells separated by a fine fibrovascular stroma. Tumor cells infiltrated the myocardium, and neoplastic emboli were present, but no metastases were macroscopically detectable. Tumor cells were immunohistochemically positive for chromogranin A, for synaptophysin and, faintly, for neuron-specific enolase and negative for vimentin, cytokeratin, alpha smooth muscle actin, glial fibrillary acidic protein, thyreoglobulin, and calcitonin. Based on histologic and immunohistochemical findings, the diagnosis of chemodectoma was made.

Effects of iloprost on adhesion molecules and F1 + 2 in peripheral ischemia.

BACKGROUND: Iloprost has beneficial effects on microcirculation by preventing platelet and leukocyte reciprocal activation, which is known to lead to endothelial damage and acute thrombosis. This drug also reduces inflammatory system activation by decreasing alpha M beta 2 integrin expression on the phagocyte membrane, might have a role in the protection and restoration of endothelial integrity and might interact with coagulation cascade activation. DESIGN: Forty patients were enrolled: 29 with systemic sclerosis (SSc) and 11 with peripheral artery disease (PAD). Iloprost was administered for 5 days in the first group and for 21 days in second group of patients. To ascertain whether iloprost modifies the parameters of endothelial and coagulation cascade activations, the plasma concentrations of S-ICAM-1 and F1 + 2 were detected in patients at baseline, after 5 days and, in PAD patients only, after 21 days of iloprost therapy. S-ICAM-1 is the endothelial counter receptor for alpha M beta 2 integrin and is a marker of endothelial cell activation; and F1 + 2 is a marker of coagulation cascade activation. RESULTS: After infusion of iloprost a significant decrease of S-ICAM-1 was observed in both the SSc (P < 0.002) and PAD patients (P < 0.004). Similarly, a significant decrease of F1 + 2 was observed in the SSc (P < 0.0004) and PAD patients (P < 0.003). CONCLUSIONS: The study provides evidence that iloprost reduces endothelial cells and coagulation cascade activations. Both of these mechanisms are responsible for improvement in microvascular functional capacity and for the long-term clinical benefit observed. After iloprost infusion, the SSc patients showed marked reductions in F1 + 2 and S-ICAM-1 concentrations that were statistically more significant relative to the PAD patients.

Treatment of hyperthyroidism due to inappropriate secretion of thyrotropin with the somatostatin analog SMS 201-995.

The management of hyperthyroidism due to inappropriate secretion of TSH (IST) includes agents that selectively suppress TSH hypersecretion both in patients with TSH-secreting tumor [neoplastic IST (nIST)] in whom pituitary surgery was unsuccessful and in those with selective pituitary resistance to thyroid hormone action [nonneoplastic IST (nnIST)]. Among such agents, somatostatin administration has proven to be effective in blocking TSH hypersecretion, but its short plasma half-life prevented its use in long term therapeutic trials. The recent availability of a potent and long-acting analog of somatostatin (SMS 201-995, Sandostatin) prompted us to study its effects on serum TSH, alpha-subunit, and free thyroid hormone (FT4 and FT3) concentrations in five patients with nIST and three patients with nnIST. During short term SMS 201-995 administration (100 micrograms, sc, three times daily for 5 days) both serum TSH and alpha-subunit levels decreased in all patients with nIST (mean decrements, -86% and -85%, respectively), with concomitant normalization of serum FT4 and FT3 concentrations. In the three patients with nnIST, this treatment lowered serum TSH levels less well (mean decrement, -47%), although serum FT4 and FT3 levels normalized in one patient. Chronic SMS 201-995 (100 micrograms, sc, every 12 h for 1-7 months) treatment in four hyperthyroid patients (two with nIST and two with nnIST) resulted in a steady euthyroid state in both patients with nIST, with restoration of normal visual fields in one patient. In contrast, in both patients with nnIST, escape occurred after 2 weeks of therapy. We conclude that SMS 201-995 administration is effective treatment for patients with nIST, able to suppress TSH hypersecretion from the adenomatous thyrotrophs and, consequently, to restore clinical and biochemical euthyroidism in such patients. On the contrary, the inhibitory effects of SMS 201-995 on TSH secretion in patients with nnIST are weaker and transient.

Treatment of inappropriate secretion of thyrotropin with somatostatin analog SMS 201-995.

Inappropriate thyrotropin secretion (IST) may originate from either neoplastic disease (nIST) or non-neoplastic resistance to thyroid hormone (nnIST). An inhibitory effect of somatostatin on TSH secretion has been documented. In an attempt to elucidate the possible therapeutic effect of this peptide on nIST and nnIST, a study was conducted in 7 such patients. Sandostatin (SMS 201-995) was administered in daily doses of 100 micrograms for several days to 1 month. Four patients with nIST responded with a fall in circulating TSH as well as alpha-subunit with concomitant normalization of free thyroxine and clear symptomatic improvement. In the 3 nnIST patients this effect was considerably less apparent and a partial TSH escape was observed on long-term treatment in 2 cases. The importance of somatostatin and its analogs in the management of thyroid malignancy is stressed.

Study of the effects of bromocriptine on sexual impotence.

Serum testosterone, prolactin, LH, FSH and plasma 17 beta-oestradiol levels were determined in forty-seven male patients with sexual impotence. Low testosterone values and slightly elevated prolactin levels were observed in 19% and in 17% of cases, respectively. Since sexual function in the male seems to be controlled by both dopaminergic stimulatory and serotoninergic inhibitory mechanisms, bromocriptine was given orally to seventeen patients. As the preliminary results appeared encouraging, a double blind study was undertaken in the other thirty patients. No appreciable difference in hormonal pattern was noticed between bromocriptine and placebo treatment. As far as sexual function was concerned, good results were obtained in 52% of cases treated with bromocriptine and in 44% of patients given placebo.

Effect of two serotonin antagonists on prolactin and thyrotrophin secretion in man.

The effects of serotoninergic blockade on prolactin and thyrotrophin secretion in man was evaluated by determining the basal and TRH-stimulated serum prolactin and TSH concentrations in normal volunteers before and after a 3 days course of cyproheptadine or methergoline administration. Cyproheptadine, a serotonin antagonist with antihistaminic, anticholinergic and antidopaminergic properties as well, did not affect prolactin secretion, while it reduced the serum TSH response to TRH; methergoline, a specific blocker of central serotonin receptors, decreased basal and TRH-induced serum prolactin levels, without affecting TSH secretion. These results support the existence of serotoninergic stimulatory influences on human prolactin release, while suggesting that human TSH secretion is not modulated by serotoninergic inputs.

Failure of oral 5-hydroxytryptohan administration to affect prolactin secretion in man.

Oral 5-hydroxytryptopahn (5-HTP) administration, either 200 mg acutely or 50 mg q.i.d. fr 3 days plus 200 mg acutely, failed to modify either basal or TRH-stimulated prolactin secretion in normal subjects. This is at variance with the stimulatory action of intravenous tryptophan on human prolactin release. However, it is doubtful that 5-HTP at the dose used may increase brain serotonin concentration; moreover, the hydroxlated amino acid is also taken up by catecholaminergic neurons, from which noradrenaline and dopamine may then be released. It is concluded that the failure of 5-HTP to affect prolactin secretion in man is not a proof against the existence of serotoninergic stimulatory influences on human prolactin release.