Comparison between May-Grünwald-Giemsa and rapid cytological stains in fine-needle aspirates of canine mast cell tumour: Diagnostic and prognostic implications.

Mast cell tumours (MCTs) are often diagnosed by cytology based on the identification of purple intracytoplasmic granules with methanolic Romanowsky stains, including May-Grünwald-Giemsa (MGG). In clinical practice, aqueous rapid stains (RS) are commonly used, but mast cell granules may not stain properly. Aim of this prospective study was to investigate the frequency of MCT hypogranularity with RS and its potential implications in tumour identification, cytological grading assessment and recognition of nodal metastatic disease. Cytological preparations of canine primary MCTs and metastatic lymph nodes with subsequent histopathological confirmation were included. For each case, good-quality smears were stained with both MGG and RS and comparatively assessed. Eleven of 60 (18.3%) primary MCTs were hypogranular with RS; 9 of them were histologically high-grade tumours and in 3 cases (5%) a definitive MCT diagnosis could not be made. Accuracy in cytological grading assessment (85%) did not differ between RS and MGG. Thirteen of 28 (46.4%) metastatic lymph nodes were hypogranular with RS and 3 independent observers failed to identify nodal MCT metastases in 7% to 18% of RS-stained smears. This study confirms that, in limited cases, RS can be ineffective in staining MCT granules, particularly in high-grade tumours, thus making diagnosis more dependent on experience and quality of preparations. In dubious cases, methanolic stains should be applied. The use of RS is discouraged for the search of nodal metastases, as the identification of isolated mast cells can be more challenging.

Evidence for bovine besnoitiosis being endemic in Italy--first in vitro isolation of Besnoitia besnoiti from cattle born in Italy.

Until 2009, bovine besnoitiosis had never been considered endemic in Italy and the only report on the disease in this country referred to animals imported from France shortly before. However, recently, an autochthonous outbreak of bovine besnoitiosis was reported in four herds located at the intersection of the borders between Emilia-Romagna, Toscana and Marche (Northern Apennine Mountains), which has led to an increased awareness concerning this disease. The present study describes a further outbreak of bovine besnoitiosis in Italy. The afflicted herd was a dairy herd with no evidence for contact with cattle from regions known to be endemic for bovine besnoitiosis. The farm investigation was initiated after a three-year old Holstein Friesian dairy cow with generalized thickening and lichenification of the skin was diagnosed with bovine besnoitiosis. The clinical diagnosis was confirmed by gross pathology, histopathology, serology and PCR. Bradyzoites released from tissue cysts obtained from the skin of this animal enabled the first in vitro isolation of Besnoitia besnoiti in Italy. This isolate was named Bb-Italy1. Sequencing of a 2118 bp spanning region including the complete internal transcribed spacer 1 and parts of the 18S and the 5.8S rRNA gene from DNA extracted from skin-derived zoites revealed a 99.9% identity to sequences known for other B. besnoiti isolated from cattle in Europe. Two GKO mice which had been inoculated intraperitoneally with bovine skin-derived bradyzoites became ill 7 days post inoculation. Parasitophorous vacuoles with multiplying zoites were observed in the cell culture inoculated with peritoneal fluids of these mice and a B. besnoiti infection in the mice and in the cell culture could be confirmed by real-time PCR. A serological investigation in the afflicted herd using immunoblots and an immunofluorescent antibody test (IFAT) revealed an overall herd seroprevalence of 9.7% (31/321), whereas within the female animals older than 2 years 17.0% (29/171) of the dams were tested positive. With one exception, an imported cow from Germany, all the seropositive animals were born in Italy. In connection with previously described autochthonous cases of bovine besnoitiosis the case described herein suggests that bovine besnoitiosis should be considered endemic in Italy.

Evidence for bovine parafilariosis in Italy: first isolation of Parafilaria bovicola (Tubangui, 1934) from autochthonous cattle.

In May and July 2010 and in March 2011, the presence of multiple bleeding nodules at the level of the neck, shoulders, withers, back and rump in five cattle from three herds in north-central Italy were observed. A diagnosis of parafilariosis was made. Fragments of adult females of Parafilaria bovicola could be identified from the derma of three to five bioptic extirpates. Larvated eggs and free larvae were observed in serohemorrhagic exudates. This paper is the first report of the occurrence of parafilariosis in Italy and subsequent to the various outbreaks reported, it can now be said that bovine parafilariosis is also currently present in Italy.

Cytological diagnosis of mandibular salivary gland adenocarcinoma in a dog.

A case of mandibular salivary gland adenocarcinoma in a 9-year-old female dog is described. Material collected by fine needle aspiration underwent cytological examination and after the diagnosis of salivary carcinoma the mass was surgically excised and then was processed for histological examination. The aim of this work is to describe the cytopathological features of this carcinoma and to emphasize the usefulness of fine needle aspiration technique, which is an effective, inexpensive and minimally invasive method of diagnosis that can be performed before incisional biopsy or even before surgical excision.

Different biological behaviour of Waldenström Macroglobulinemia in two dogs.

Waldenström Macroglobulinemia is a low-grade immunosecretory disorder associated with lymphoid tumours, which is rarely reported in veterinary medicine. In this study, we describe two clinical cases of this rare syndrome in dogs, each characterized by a different onset and clinical course. In one case, a hyperacute onset and aggressive behaviour of the neoplasm was observed. Absolute serum viscosity (SV) was retrospectively evaluated in order to explain clinical findings. Rotational viscosimetry showed good precision in measuring SV. Both dogs had SV values higher than a control groups of healthy dogs although only one subject developed hyperviscosity symptoms and complications. At high paraprotein concentrations, a slight reduction of the M-component was associated with a marked decrease in SV. Thus, this work suggests that SV assessment is a relevant tool for managing monoclonal gammopathies, whose usefulness should be further confirmed in larger cohorts of dogs.

Transitional cell carcinoma of the renal pelvis in two dogs.

Transitional cell carcinoma (TCC) of the renal pelvis was found in two dogs, a 7-year-old male English Setter and a 11-year-old female Shetland shepherd. Affected dogs were presented for clinical examination without any specific symptoms but haematuria in case 1 and occurrence of whitish material in the urine of case 2; neoplastic disorders were discovered with ultrasonographic investigation and fine needle aspiration biopsy. Histopathological examination was carried out after nephrectomy and ureterectomy of the affected kidney of both dogs, and confirmed the diagnosis of non-invasive and low grade TCC in case 1 and of infiltrating TCC in case 2. The clinical, gross, cytological and histopathological features of these rare tumours originating from transitional epithelium of the renal pelvis are reported.

Late biliary tract complications after orthotopic liver transplantation: diagnostic and therapeutic role of endoscopic retrograde cholangiopancreatography.

BACKGROUND: Biliary tract complications are frequent after orthotopic liver transplantation. Late biliary tract complications occurring after T-tube removal mostly include stones and strictures which may be associated with sepsis and worsening of the liver function. Endoscopic retrograde cholangiopancreatography (ERCP) has a role in the diagnosis and therapy of these complications. The aim of our study was to report our experience of endoscopic diagnosis and treatment of late biliary tract complications in liver-transplanted patients. METHODS AND RESULTS: One hundred and thirty-six adult liver-transplanted patients have been followed since 1988. Seventeen patients (12.5%) needed a total of 30 ERCP because of evidence of clinical and/or biochemical cholestasis: eight with biliary stricture; six with biliary stones; one with both stricture and stones; and two with normal ERCP findings. Interventional endoscopic procedures included 14 sphincterotomies, six stone removals, seven biliary balloon dilatations, seven biliary stent placements, 11 biliary stent replacements, seven nasobiliary catheter placements and one mechanical lithotripsy. No complications were seen. In all cases, ERCP was able to identify the location, entity and dimension of the late biliary tract complication, thus allowing a therapeutic strategy to be used. Two patients had medical cholestasis. Forty-seven per cent of patients with late biliary tract complications could definitely be cured by ERCP alone. The ERCP improved the patients' condition to allow subsequent surgery in five patients (33%). CONCLUSIONS: These results confirms that ERCP is a valuable diagnostic tool and should be considered as the first step in the non-surgical management of late biliary tract complications after orthotopic liver transplantation.

Prolongation of discordant heart xenograft survival after liver hemoperfusion: role of nonparenchymal liver cells.

In the discordant guinea pig (GP)-to-rat combination, heart xenografts are hyperacutely rejected. The aim of the present study was to demonstrate that a donor-species-specific extracorporeal liver hemoperfusion can prolong survival of discordant heart xenografts and to specify the role of non-parenchymal cells. GP hearts were grafted into Brown Norway rats (group 1) In group 2, heart xenografting was carried out immediately after a 15-min GP hemoperfusion. In group 3, Kupffer cells of the GP liver were blockaged by intravenous injection of dextran sulfate (4 mg/100 g) 30 min before hemoperfusion. In group 4, Kupffer cells of the liver were activated by intravenous injection of muramyl dipeptide (MDP; 500 micrograms/250 g) 24 h before hemoperfusion. Lymphocytotoxic antibodies were detected according to a complement-dependent antibody assay. A donor-specific liver hemoperfusion can delay hyperacute rejection of heart xenografts (67.6 +/- 47.1 min in group 2 versus 8.0 +/- 2.4 min in group 1; p < 0.01) and reduce the level of lymphocytotoxic antibodies. Deposits of immunoglobulins and complement were significant on the hemoperfused liver and moderate on the transplanted heart. In group 3, after blockade of Kupffer cells with dextran, heart xenograft survival was less prolonged (31.8 +/- 8.2 min) and the decrease in antibody levels was not significant and associated with moderate deposits of immunoglobulins and complement on the hemoperfused liver and significant deposits on heart xenografts. In group 4, after stimulation of Kupffer cells by MDP, a significant decrease in antibody levels was present, and significant deposits were observed. These results show that donor-specific liver hemoperfusion can prolong the survival of discordant heart xenografts and support the hypothesis that nonparenchymal liver cells play a major role by absorption of preformed antibodies and complement.

[Orthotopic liver transplantation and hepatocellular carcinoma]. / Trapianto ortotopico di fegato e carcinoma epatocellulare.

Nineteen orthotopic liver transplantations (OLT) were performed in patients with hepatocellular carcinoma, between March 1988 and December 1990, in our Department. Thirteen patients (68.4%), 10 men and 3 women, mean age 48.2 +/- 2.8 years, were clear cut neoplastic disease, six patients (31.6%), 4 men and 2 women, mean age 48.6 +/- 4.2 years, were incidental tumors. Three years survival rate was 20% in clear cut neoplastic diseases (4 patients died for neoplastic recurrence) and 66% in incidental tumors. Our results demonstrate that liver transplantation for hepatobiliary malignancy is still justified, patient selection is important in predicting outcome.

[Recurrence of virus B hepatitis after liver transplantation]. / Récidive du virus de l'hépatite B après transplantation hépatique.

Thirty-three HBs antigen positive patients without signs of viral replication underwent orthotopic liver transplantation and received long term passive immunoprophylaxis with anti-HBs immunoglobulins at high doses perioperatively and then at a dose of 10,000 IU every month. All patients became negative during the first 6 months following surgery. At 34 months the survival rate was 67 percent and the actuarial recurrence rate of serum HBs antigen was 7.1 percent. Reappearance of HBs antigen was associated with evidence of HBV replication and histological alterations of the graft. In our experience, long term passive immunoprophylaxis reduces HBV reinfection of the grafted liver.

Protective effects of prostaglandin E1 on normothermic liver ischemia.

Effects of treatment with prostaglandin E1 (PgE1) on normothermic liver ischemia were studied in male Lewis rats. Animals were subjected to 90 min of warm liver ischemia. Two groups of rats were constituted: group A (no treatment) and group B (PgE1 treatment). PgE1 (100 micrograms/kg) was given as a bolus 2 min before induction of ischemia and 2 min before the end of ischemia. Survival rates were assessed and, 6 h after the end of ischemia, serum transaminases, histology of the liver, Kupffer cell activity were evaluated. PgE1 treatment significantly improved survival rate (80%) in comparison with the nontreated group (40%). A significant reduction in transaminase levels was observed after PgE1 The extent of necrosis and congestion was improved by PgE1 treatment. Sheep red blood cell 51Cr liver uptake was deeply depressed 6 h after the end of ischemia in group A (6 +/- 2.3%/g tissue), and was significantly higher (p less than 0.001) after PgE1 administration in group B (32.98 +/- 11.7%/g tissue). Our results demonstrate that PgE1 is able to protect the liver from ischemic insult. The mechanism by which prostaglandins exert this beneficial effect on normothermic liver ischemia may be related to their action on hepatic macrophages.