Granular Cell Tumour in a California Kingsnake (Lampropeltis californiae).

A 21-year-old female California kingsnake (Lampropeltis californiae) was presented to the University of Veterinary Medicine, Vienna, Austria, with a space-occupying mass in the caudal abdomen. Following clinical, radiological and sonographical evaluation the mass was removed surgically. Histopathology and transmission electron microscopy confirmed the diagnosis of a granular cell tumour, but immunohistochemical labelling for a range of markers was negative. This lesion is rare in mammals and birds, but has not been reported previously in a reptile.

Pax5 immunostaining in paraffin-embedded sections of canine non-Hodgkin lymphoma: a novel canine pan pre-B- and B-cell marker.

The Pax5 gene encodes the B-cell specific activator protein (BSAP), a member of the highly conserved paired box (PAX)-domain family of transcription factors and a key regulator in the development and differentiation of B-cells. Pax5 serves as a valuable B-cell marker in the classification of human lymphoma patients as it is restricted to lymphomas of B-cell lineage. In dogs, detection of Pax5 protein in lymphoma tissue has not been reported. Therefore, we have investigated the expression and detection of BSAP using a monoclonal anti-Pax5 antibody (anti-BSAP, clone 24) in canine lymphoma tissue samples to evaluate its diagnostic relevance as a B-cell marker. A series of 25 lymph nodes from 23 canine non-Hodgkin lymphoma patients, a reactive canine lymph node, and a normal non-reactive canine lymph node, were evaluated. All B-cell non-Hodgkin lymphomas (15) were found to express Pax5 protein. In addition, there was a strong correlation between Pax5 and CD79a expression. Three CD3 positive and five CD3 and CD79a positive lymphomas were immunophenotypically negative for anti-Pax5, indicating a T-cell lineage. In conclusion, anti-Pax5 antibody may offer an excellent B-cell marker in canine lymphomas.

IL-4 downregulates expression of the target receptor CD30 in neoplastic canine mast cells.

CD30 is a novel therapeutic target in human mast cell (MC) neoplasms. In this 'comparative oncology' study, we examined CD30 expression and regulation in neoplastic canine MC using a panel of immunomodulatory cytokines [interleukin-2 (IL-2), IL-4, IL-5, IL-6, IL-13 and stem cell factor (SCF)] and the canine mastocytoma cell lines NI-1 and C2. Of all cytokines tested IL-4 was found to downregulate expression of CD30 in NI-1 and C2 cells. We also found that the CD30-targeting antibody-conjugate brentuximab vedotin induces growth inhibition and apoptosis in both MC lines. Next, we asked whether IL-4-induced downregulation of CD30 interferes with brentuximab vedotin-effects. Indeed, pre-incubation of NI-1 cells with IL-4 decreased responsiveness towards brentuximab vedotin. To overcome IL-4-mediated resistance, we applied drug combinations and found that brentuximab vedotin synergizes with the Kit-targeting drugs masitinib and PKC412 in inhibiting growth of NI-1 and C2 cells. In summary, CD30 is a new marker and IL-4-regulated target in neoplastic canine MC.

Pyogranulomatous panniculitis in ferrets (Mustela putorius furo) with intralesional demonstration of Pseudomonas luteola.

One ferret (Mustela putorius furo) from Finland and two ferrets from Austria, aged 1-4.5 years and of both genders, were presented with pyogranulomatous subcutaneous inflammation affecting the inguinal, preputial and femoral regions, respectively. Histologically, microorganisms were detected within the lesions. The organisms had a capsule that stained positively by the periodic acid-Schiff reaction. Pseudomonas spp. were cultured from the lesions in two cases. In the third case, electron microscopy revealed a prokaryotic organism surrounded by an electron lucent matrix. 16S rRNA gene sequencing showed highest sequence homology to Pseudomonas luteola in all three cases. In combination with recent reports of pleuropneumonia in ferrets due to P. luteola infection, these cases might indicate a predisposition of ferrets for infection by these bacteria.

Cytogenetic studies in a canine mammary tumor.

In a 9-year-old female dog (Basset Artesian Normand) with a mammary adenocarcinoma, cytogenetic evaluation of tumor cells showed a chromosome number of 76 in most metaphases (95%). The following abnormalities were found: symmetric metacentric chromosomes 1 and 6, centric fusion 3/38, a marker X-chromosome (Xmar) and a biarmed small marker chromosome (mar). In the remaining metaphases (5%) there were additional biarmed marker chromosomes present.

Loss of chromosome B2-material in three cases of feline mammary tumours.

Tumour cells in three cases of mammary gland neoplasms in cats were analysed cytogenetically. There was a loss of chromosome B2-material in all the cases, and a loss of chromosome E3-material in two of them.

Chromosomal hyperdiploidy in a feline sarcoma.

An eight-year-old male cat developed a sarcoma. The cytogenetic evaluation of the tumour cells showed the presence of hyperdiploidy (range 40 to 46 chromosomes). This hyperdiploidy was encountered in all the cells examined. Extra-chromosomes numbers C1, C2, B4, D4 and E3 were mainly responsible for the hyperdiploid chromosomal complements. There was a high incidence of monosomy E3.

Trisomy D2 in a feline neurofibroma.

An eight-year-old female cat developed a skin neurofibroma. The cytogenetic evaluation of the tumour cells showed the presence of a high percentage (16.4 per cent) of trisomic cells. The trisomy concerned chromosome number D2.

Cytogenetic analyses of four solid tumours in dogs.

Four solid tumours (one haemangiopericytoma, one haemangioendothelioma, one spindle-cell sarcoma and one mammary carcinoma) in dogs were analysed cytogenetically. In the haemangiopericytoma, an additional small chromosomal segment was present. Very complex changes including centric fusions and symmetric meta-centrics 1, 6, 10 and 12 were conspicuous in the highly unbalanced karyotype of the haemangioendothelioma. Complex changes, particularly many centric fusions and a tandem translocation 4/14, were features of the spindle-cell sarcoma. One centric fusion and a symmetric metacentric 13 were present in the mammary carcinoma.

Cytogenetic variation between four cases of feline fibrosarcoma.

Short term cultures of four feline fibrosarcomas were analysed cytogenetically. There was marked genetic heterogeneity between the four cats, each showing a different clonal abnormality. The aberrations detected were one deleted B2, one marker F1 and two reciprocal translocations, t (A2q; E3q) and t (A1q; B4p).

Presence of p53 mutations in feline neoplasms.

A region from exon 4 to 8 of the tumour suppressor gene p53 was analysed in 60 feline tumours (30 fibrosarcomas, seven malignant histiocytomas, three lymphosarcomas, five basal cell tumours, five squamous cell carcinomas, two adenocarcinomas of tubular skin glands, one undifferentiated carcinoma of the skin, seven mammary carcinomas). Missense mutations were detected in two fibrosarcomas, one malignant fibrous histiocytoma, the undifferentiated carcinoma of the skin and one mammary carcinoma. One nonsense mutation was detected in one fibrosarcoma and one deletion/frameshift-mutation was observed in one squamous cell carcinoma.

Cytogenetic alterations in eight mammary tumors and tumor-suppressor gene p53 mutation in one mammary tumor from dogs.

OBJECTIVE: To identify mutations in canine mammary tumors. ANIMALS: 10 tumor-bearing dogs. PROCEDURE: Culture of neoplastic cells originating from mammary tumors was performed, and trypsin-G banding was used for cytogenetic investigations. The same tumors were subjected to molecular genetic screening by use of DNA extraction, polymerase chain reaction, DNA elution, and DNA sequencing for ras oncogenes and the p53 tumor suppressor gene. RESULTS: A broad spectrum of chromosome aberrations was observed, including trisomies, reciprocal translocations, and structural and numerical X-chromosome alterations and deletions. Molecular genetic analysis revealed a tumor suppressor p53 gene mutation in codon 249 of exon 7 in one instance. Interestingly, analyzed mammary tumors were free of mutations in N-ras, K-ras and H-ras, exons 1 and 2. CONCLUSIONS: Chromosome alterations are wide-spread in canine mammary tumors, but no ras family mutations were detected in tumors from these 10 dogs. CLINICAL RELEVANCE: Knowledge about chromosome, oncogene, and tumor suppressor gene damage could be helpful for diagnosis and prognosis of neoplastic diseases in dogs.

Lectin binding patterns of uterine glands in mares with chronic endometrial degeneration.

OBJECTIVE: To evaluate changes of glycoconjugate in uterine glands of endometrial tissues obtained from mares. ANIMALS: adult mares. PROCEDURE: Uterine biopsy samples were collected during the breeding season and analyzed histologically for signs of chronic endometrial degeneration. Stage of the estrous cycle was established, using clinical examination and determination of hormonal status. Uterine tissue samples were analyzed, using lectin histochemical and immunohistochemical techniques (estrogen and progesterone receptors). Connective tissues were stained to determine alterations of ground substance in periglandular fibrosis. RESULTS: Of 50 mares, 30 (60%) were classified as normal or having modest alterations, and 20 (40%) were classified as having moderate or severe endometrial degeneration. In normal equine endometrium, several lectins (Helix pomatia agglutinin, Lotus tetragonolobus agglutinin, Ricinus communis I agglutinin, Ulex europaeus agglutinin, and wheat germ agglutinin) bound to glycoconjugates of the luminal epithelium and openings of uterine glands. Lectin binding patterns of cystic dilated glands or fibrotic glands in endometrial samples were remarkably strong, whereas normal surrounding cells remained unstained. Lotus tetragonolobus lectin was not suitable for detecting endometrial alterations. Connective tissues stained with Alcian blue and results of Hale colloidal-iron binding revealed acidic ground substance in periglandular fibrosis. Estrogen and progesterone receptors were evenly distributed in healthy and affected endometrial samples. CONCLUSIONS AND CLINICAL RELEVANCE: Glycoconjugate patterns of uterine glands were altered in mares with chronic endometrial degeneration. Therefore, uterine secretions are likely to be altered. These changes are not induced by changes in content of estrogen and progesterone receptors in endometrial tissues.

Novel canine tumour suppressor gene p53 mutations in cases of skin and mammary neoplasms.

Exons 4 to 8 of the tumour suppressor gene p53 were analysed in 25 skin and 25 mammary tumours of 50 dogs. A 1 bp deletion (ACC-->AC) was detected in codon 89 in exon 4 in a squamous cell carcinoma. A missense mutation CGC-->CAC (arginine-->histidine) was present in codon 162 in exon 5 in a mammary adenocarcinoma. Moreover, a silent mutation occurred in codon 103 (serine) of exon 4 in a mammary adenoma. The somatic nature of the three mutations was demonstrated.

Cytogenetic alterations in four feline soft-tissue tumours.

Four mesenchymal neoplasms (two malignant fibroblastic histiocytomas, one fibrosarcoma, one fibroma) were investigated cytogenetically. Although no specific alterations were present, the frequent nonrandom involvement of chromosome E1 became clearly evident. Karyotypic alterations involving unrelated clonal changes were also present in the fibroma.

Absence of p21 WAF1 and p27 kip1 gene mutations in various feline tumours.

The coding regions of tumour suppressor and cell cycle regulatory genes p21 WAF1 and p27 Kip1 were investigated in 101 feline tumours of various types. No damaging mutations were present in the analysed areas of the genes.

Novel p53 tumour suppressor mutations in cases of spindle cell sarcoma, pleomorphic sarcoma and fibrosarcoma in cats.

Twenty feline neoplasms were sequenced in the region from exons 5 to 8 for the presence of tumour suppressor gene p53 mutations. In a spindle cell sarcoma of the bladder, a missense mutation (codon 164 AAG-->GAG, lysine-->glutamic acid) in exon 5 was detected. In a pleomorphic sarcoma, a 23 bp deletion involving the splicing junction between intron 5 and exon 6 was observed. In a fibrosarcoma, a 6 bp deletion of p53 covering 2 bp of exon 7 and 4 bp of intron 7, including the splicing junction, was found. The study demonstrates three new p53 mutations in different types of sarcomas in cats.

Canine tumour suppressor gene p53--mutation in a case of adenoma of circumanal glands.

Highly conserved regions of the tumour suppressor gene p53, including the typical human tumour hot spots (codons 175, 245, 248, 249, 273 and 282), were investigated in various canine neoplasms. A mutation CGG-->TGG (arginine-->tryptophan) was detected in codon 249 in an adenoma of the circumanal gland.

Canine tumour suppressor gene p53 mutation in a case of anaplastic carcinoma of the intestine.

Tumours localised in the large bowel of dogs were subjected to molecular genetic studies. Highly conserved regions of the tumour suppressor gene p53, including typical tumour hot spots (codons 175, 245, 248, 249, 273 and 282), were analysed. A mutation CGG-->TGG (arginine-->tryptophan) was present in codon 249 in an anaplastic carcinoma in the caecum.

Tumour suppressor gene p53 mutation in a case of haemangiosarcoma of a dog.

Haemangiosarcomas of dogs were analysed by molecular genetic techniques. Regions of the tumour suppressor gene p53, including the well-known tumour hot spots (codons 175, 245, 248, 249, 273 and 282) were screened. A 24 bp deletion was detected in exon 5 of the gene.