Oxidative stress parameters in bitches with mammary gland tumours

The aim of the present study was to describe some of the oxidative stress parameters in bitches suffering from spontaneously occurring mammary gland tumours. The experiment involved 28 bitches which had mammary gland tumours removed surgically (15 bitches with malignant tumour and 13 with benign tumour) as well as 10 clinically healthy bitches. The activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were determined in haemolysates of erythrocytes derived from the animals. The concentrations of thiobarbituric acid reactive substances (TBARS), as well as -SH groups, were determined in blood plasma. GSH-Px activity was significantly higher in the malignant tumour group than in healthy animals. SOD activity was significantly higher in animals with tumours compared with the control group. Activities of both enzymes were higher in animals with malignant tumours than in benign groups, but the differences were not statistically significant. The concentrations of TBARS and -SH groups were similar in all examined groups. The increase of antioxidative enzyme activities in these animals may suggest the activation of antioxidative defence mechanisms in mammary gland carcinogenesis. Moreover, it might indicate the participation of oxidative stress in malignancies. Further experiments involving more animals, with more frequent sample collection and the use of other oxidative stress markers are necessary.     

An assessment of the clonality of the components of canine mixed mammary tumours by mitochondrial DNA analysis

The aim of this study was to investigate if mutations in the mitochondrial DNA (mtDNA) D-loop fragment control region of canine mammary mixed tumours could be used as clonal markers that identified the cell population of origin. Ten benign mixed mammary tumours and nine carcinomas arising from benign mixed tumours were microdissected and DNA from epithelial and mesenchymal tumour cells and from normal mammary tissue was examined for sequence variations in a fragment of the hypervariable control region.Identical sequence variants in both the epithelial and mesenchymal components (as well as in the corresponding normal tissue) were found in 80% of the benign mixed tumours and in 89% of the carcinomas arising from benign mixed tumours suggesting a shared clonal origin. The distinctive sequence alterations identified in the epithelial and mesenchymal components of 15.8% of all 19 tumours examined, suggests the possibility that a minority of mammary tumours are polyclonal in origin or that early clonal divergence occurs. Increased mutation within the mtDNA D-loop fragment of mixed tumour components was not observed.     

Expression and significance of PTEN in canine mammary gland tumours

To explore the expression and clinical importance of the anti-oncogene phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in canine mammary gland tumours, PTEN expression was compared in 50 cases of canine mammary tumour and four examples of normal mammary tissue using real-time quantitative PCR. PTEN expression was similar in benign mammary tumours and normal mammary tissues (P>0.05), but was lower in malignant tumours than in normal mammary tissues or benign mammary tumours (P<0.001). PTEN expression was also low in the lymph node metastases of malignant mammary tumours. The expression profile of PTEN in malignant mammary tumours compared to those without lymph node metastasis varied significantly. Low-level PETN expression might play an important role in carcinogenesis and the progression of canine mammary tumours, and PTEN protein detection might be useful in evaluating tumour development and prognosis.     

Effects of different liver diseases on the platelet count in dogs

Changes of the platelet count in liver diseases are described in humans. Thrombocytopenia was observed more frequently than thrombocytosis. There are only a few investigations on platelet counts in liver diseases in dogs. The goal of the present study was to investigate the influence of different liver diseases including degeneration, hepatitis and liver tumours, on the platelet count. Platelet counts of 52 dogs with different liver diseases were measured and compared with 52 healthy dogs. The results showed, that dogs with liver degeneration have thrombocytosis in 41% of the cases and a group of dogs with liver tumours (malignant histiocytosis, hepatoma, malignant lymphoma anaplastic sarcoma, cholangiocarcinoma, hepatocellular carcinoma) had thrombocytopenia in 50% of the cases. The dogs with hepatitis showed no specific changes in the platelet count. The statistical comparison of our patients with liver disease and a control group of healthy dogs showed significantly higher platelet counts in cases of liver degeneration (p < 0.0001) and significantly lower platelet counts in cases of liver tumour (p < 0.001). The comparison between the dogs with different liver diseases showed significantly lower platelet counts in dogs with liver tumours when compared to dogs with liver degeneration (p < 0.0001). There was no significant difference between dogs with liver tumours and dogs with hepatitis and between dogs with liver degeneration and dogs with hepatitis. Based on the results of this study the author recommends to assess platelet counts in all dogs with liver disease, especially if liver biopsy is planed.     

p53 expression and apoptosis in melanomas of dogs and cats

Twenty-seven melanocytic tumours from 20 dogs and four cats were examined for p53 expression and apoptosis. They included tumours that were histologically classified as benign (BM), primary malignant (PMM) and metastatic malignant melanomas (MMM). For all cases clinical follow-up was available. p53 expression was examined immunohistochemically using different monoclonal and polyclonal antibodies. Apoptosis was detected using the TUNEL technique. The tissue sections were analysed using a quantitative image analysing system. A p53 index (p53I) and an apoptotic index (AI) were determined. p53 over-expression was found infrequently in these canine and feline melanocytic tumours. Apoptosis was observed in some of the malignant tumours. In one feline case of malignant melanoma, p53 accumulation together with apoptosis was seen in three metastases but not in the primary tumour. p53I and AI were not significantly correlated with survival. These results are similar to those reported for human cutaneous melanomas.     

Hepatic alveolar echinococcosis in cynomolgus monkeys (Macaca fascicularis)

Alveolar echinococcosis was diagnosed in 12 cynomolgus monkeys (Macaca fascicularis) at postmortem examination within a period of 6 years. Besides consistent involvement of the liver, parasitic lesions were also present in mesenteric lymph nodes, pancreas, lung, and kidney. In the liver, various patterns of host's responses to parasitic tissue could be distinguished. Infiltration of macrophages, often multinucleated, around usually intact metacestodes was the main feature of one pattern. A second pattern was characterized by the presence of abundant, normally degenerate granulocytes in addition to macrophages surrounding collapsed laminated structures. Finally and as a third pattern, some cysts were surrounded by marked collagen deposition, which was usually not a significant feature of the other foci. Parasitic cysts with protoscolices were observed in foci with the first and third pattern but not in the second one. The simultaneous occurrence of all three patterns was observed in most animals. Type AA amyloid was identified either in the space of Dissé, macrophages or blood vessel walls in nine animals using immunohistochemistry. Identity of parasitic structures such as metacestodes of Echinococcus multilocularis was confirmed immunohistochemically. All animals that could be tested serologically (7/12) had detectable antibodies against the E. multilocularis-specific Em2 antigen. Liver lesions of six animals were additionally analyzed by polymerase chain reaction, yielding the amplification of a specific E. multilocularis DNA fragment in each case.     

Expression of TopBP1 in canine mammary neoplasia in relation to histological type, Ki67, ERalpha and p53

TopBP1 is aberrantly expressed in human and feline mammary carcinomas, but expression of this BRCA1-related protein has not been investigated in canine mammary carcinomas. In this study, 132 canine mammary tumours (46 benign, 86 carcinomas) were examined immunohistochemically for expression of TopBP1, oestrogen receptor α (ERα), Ki67 and p53. Positive staining for TopBP1 was evident in all canine mammary lesions, although five samples had <20% positive cells. The number of samples with high levels of staining increased in different categories from benign mixed tumour to adenoma to carcinoma. Most TopBP1 staining was nuclear, but both nuclear and cytoplasmic staining were observed as the degree of malignancy increased, similar to human and feline mammary carcinomas. Benign mixed tumours, however, had more cytoplasmic staining than adenomas. Expression of p53 and the proliferation marker Ki67 increased from benign mixed tumour to adenoma to carcinoma, but the differences between benign and malignant tumours were more distinct than for TopBP1 expression. ERα expression decreased from malignant to benign tumours, although over half of the benign mixed tumours were negative. TopBP1 was expressed in canine mammary tumours at higher levels than has been reported previously for cats, although the shift in cellular localisation with malignancy was similar.     

Ultrasonographic characteristics of soft tissue tumours in dogs

Objective   To identify and describe the ultrasonographic features of soft tissue tumours in dogs.
Procedure   Superficial soft tissue tumours of various histological types, including mast cell tumours (MCTs) and soft tissue sarcomas (STSs), were evaluated. Ultrasound was used to visualise internal characteristics of the tumour, including vascularity. Tumours were categorised according to size, shape, margin definition, tissue plane mobility, echogenicity, echotexture, acoustic shadowing or enhancement and vessel distribution. Objective measurements of intratumoural blood flow included velocities and maximal perfused cross-sectional area (fractional area). Logistic regression models incorporating a variety of data were used in an attempt to predict the histopathological type of tumours.
Results   The logistic regression model defined by the parameters echotexture, margin definition and presence of subcapsular vessels was highly predictive of MCTs (> 73%; P = 0.024). Several other trends, including a larger size for STSs and less vascularity for both MCTs and STSs, were observed, but did not reach statistical significance.
Conclusion   This preliminary study has shown the potential diagnostic value of ultrasound in differentiating soft tissue tumours. However, at present, ultrasound cannot replace biopsy and histopathological evaluation for tumour diagnosis.     

Vaginectomy and urethroplasty as a treatment for non-pedunculated vaginal tumours in four bitches

Vaginal tumours are uncommon in dogs. Previous reports have shown that 73 to 94 per cent of documented vaginal tumours are benign and pedunculated, often on narrow stalks. Some vaginal tumours are non-pedunculated. They grow in a concentric way either towards the vestibular area or towards the cervix. Their growth can locally affect the function of other organs. Urethral and rectal compression as well as local neurological disturbances have been described. This case series describes total and partial vaginectomy associated with urethroplasty as a treatment for non-pedunculated vaginal tumours in four dogs. These surgical procedures allowed complete resection of the tumour and were associated with low morbidity. Postoperative management and short term outcome are discussed. The final outcome was favourable; throughout the follow-up period, no local recurrence or metastasis was encountered in the animals.     

Defect of the mitochondrial DNA hypervariable region as a risk factor for canine mammary tumour

The aim of this study was to identify mutations in the hypervariable region of mitochondrial DNA in canine mammary tumours and to determine their association with the process of neoplastic transformation. A total of 93 biological samples, including blood as well as normal and neoplastic tissue samples from 31 dogs with diagnosed malignant canine mammary tumours were analysed. DNA extraction, amplification and sequencing of the D‐loop as well as bioinformatic and statistical analyses were performed. In the mitochondrial D‐loop sequence, 26 polymorphic loci and 5 mutations were identified. For the first time, D‐loop length heteroplasmy was detected in dogs with mammary tumours. The malignancy grade exerted no effect on the presence of nucleotide changes. A statistically significant association between the presence of mutations and polymorphisms and the size of dogs was demonstrated. The 100% frequency of length heteroplasmy may imply that this is a hotspot mutation of canine mammary tumour.     

Histochemigal Detection of Copper in the Liver from Sheep Chronically Poisoned by Copper

Continued ingestion of copper in excess of the nutritional requirement leads in all animals to its passive accumulation within the tissues, especially the liver. Up to certain levels varying greatly with the species high concentrations of copper in the liver appear to impose no physiological hardship on the animal. Above these levels there may occur a catastrophic liberation of a high proportion of the copper into the blood stream resulting in extensive hemolysis and jaundice usually followed by death. Sheep are more prone to the hemolytic crisis of chronic copper poisoning than other animal species. The hemolytic crisis of chronic copper poisoning is associated with centrolobular necrosis of the liver. The copper concentration in the liver is usually 300 p.p.m. or more (wet tissue) compared with about 50 p.p.m. in normal sheep (Simesen & Møller 1969).     

Defect in ND2, COX2, ATP6 and COX3 mitochondrial genes as a risk factor for canine mammary tumour

The aim of this study was to identify mutations in ND2, COX2, ATP6 and COX3 mitochondrial genes in canine mammary tumour, determine their association with the process of neoplastic transformation, and phenotypic traits of dogs. In total, 93 biological samples, including blood, normal and neoplastic tissue samples from 31 dogs with diagnosed malignant canine mammary tumours were analysed. DNA sequencing of genes as well as bioinformatics and statistical analyses were performed. A total of 28 polymorphic loci and 11 mutations were identified. One of the mutations was blood heteroplasmy and two of the mutations caused an amino acid change in p.N117S and p.A184T. For the first time, mutations in mitochondrial genes were detected in dogs with mammary tumours. A statistically significant association between the presence of mutations and the size and age of dogs was demonstrated. Some of these changes may imply that these are the hotspot mutations of canine mammary tumour.     

p62/SQSTM1 expression in canine mammary tumours: Evolutionary notes

Recent studies highlighted the role of autophagy as a cardinal regulatory system for homeostasis and cancer‐related signalling pathways. In this context, the deregulated expression of p62 – Sequestosome1 (p62/SQSTM1) – a protein acting both as an autophagy receptor and signalling hub, has been associated with tumour development and chronic inflammation. Multiple clinical studies test drugs targeting autophagy, and even more research is on the way to clinical trials. However, no comparative investigations have been carried out to identify adequate preclinical models to assess p62‐based medicine. In veterinary oncology the role of p62 in cancer‐related pathways has been largely ignored. We compared p62 sequences in multiple organisms and found that canine p62 significantly diverges from the humans and from other animals sequences. Then, we chart by immunohistochemistry the expression levels of p62 in canine mammary tumours. A total of 66 tumours and 10 non‐neoplastic mammary samples were examined. The expression of p62 was higher in normal tissue and adenomas than carcinomas, with lowest levels of p62 protein detected in high grade carcinomas. In all cases examined the tumour stroma appeared to be p62‐negative. Taken together our results would suggest that in dogs the association between p62 expression and cancer cells overturns that reported in human breast carcinoma, where p62 accumulates in malignant cells as compared to normal epithelium. Thus, at least in canine mammary tumours, p62 should be not considered a tumour‐rejection antigen for an anti‐cancer immunotherapy.     

Radiological assessment of 50 cases of incisive or maxillary neoplasia in the dog

This paper reviews the radiological features of 50 canine incisive or maxillary tumours and discusses the value of radiological assessment in the diagnosis and staging of these tumours. The 50 tumours examined included 21 sarcomas, 15 carcinomas, three melanomas and an assortment of benign tumours of the oral cavity. There was not any site specificity for the different histological tumour types within the upper dental arcade, although fibrosarcomas had a tendency to be maxillary whereas the squamous cell carcinomas were equally distributed between the incisive and maxillary regions. Seventy-eight per cent of fibrosarcomas, 82 per cent of squamous cell carcinoma and all three melanomas examined showed radiological evidence of bone involvement. Radiographic changes were also seen in the benign tumours. The pattern of growth of tumours correlated with the radiological changes observed. Malignant tumours showed a tendency to irregular or aggressive bone loss whereas bone production predominated in the benign tumours.     

Does the taking of biopsies affect the metastatic potential of tumours? A systematic review of reports on veterinary and human cases and animal models

Clinicians and pathologists are sporadically asked by owners whether the taking of tumour biopsies may affect the behaviour of the tumour, including its potential to metastasise. Unfortunately, systematic studies on this subject are unavailable in veterinary medicine, and the aim of this study was to estimate the risk of adverse effects of biopsy taking on tumour progression in animals. A systematic review of veterinary and human case reports and clinical studies as well as experimental animal models of biopsy-induced tumour metastasis was undertaken. There were only two veterinary case reports of needle tract metastases (NTM) following the taking of needle biopsies from urogenital and pulmonary tumours. Seventeen experimental studies found a high incidence of NTM but only a rat osteosarcoma and a hamster squamous carcinoma model showed an increased incidence of distant or regional metastases after incision or excision biopsy. In human medicine, the occurrence of NTM has been reported after the taking of biopsies from mesotheliomas (15%), melanomas (11%) and gall bladder tumours (11%), liver metastases of colon carcinomas (4%) and mammary carcinomas (4%) but an incidence of only <1% for all other tumours. Circulating tumour cells increased immediately after the taking of biopsies from human squamous cell, prostate, breast and hepatocellular carcinomas. Although no increased risk of biopsy-induced distant metastasis has been reported for any type of tumour, this is inconclusive due to a lack of non-biopsied control groups in human studies. Reports of biopsy-induced metastasis in animal tumours indicate that the taking of transcutaneous biopsies from urogenital tumours may be associated with a risk of NTM. However, there is no evidence of a general increase in risk of distant metastases in any tumour type in people or animals. The overall risk therefore appears to be negligible when compared to the valuable information obtained from biopsies in veterinary practice.     

Studies on canine mammary tumours. I. Age, seasonal and breed distribution.

The incidence as well as age, seasonal and breed distribution of canine mammary tumours (n = 521) were studied at the Clinic of Obstetrics and Gynaecology of the University of Veterinary Science, Budapest, between 1985 and 1989. In 39 cases of mammary tumour, blood plasma oestradiol (E2) and progesterone (P) concentrations were also determined. Of all dogs referred to the clinics of the University in 1985, 0.7% had mammary tumour. On the average, 104 +/- 9.3 cases of mammary tumour were recorded at the Clinic of Obstetrics per year. This number did not increase after the Chernobyl atomic reactor catastrophe of 1986. The age distribution of canine mammary tumour found in this study shows good agreement with earlier data of the literature: mammary tumour showed the highest incidence in 10 years old dogs. The incidence of mammary tumour kept increasing with age until the 14th year of life (as expressed in per cent of animals of identical age). The number of mammary tumours was markedly higher in the spring (April-May) and autumn (September). This seasonality was demonstrable in 11 to 16 years old bitches, too. On the basis of the blood plasma E2 and P profiles, 61.5% of the clinically anoestrous animals were found to be cycling. The strikingly high ratio of pulis among dogs with mammary cancer was suggestive of a breed disposition.     

细粒棘球蚴感染羊肝脏包囊纤维化形成的病理形态学观察

为了探讨细粒棘球蚴感染羊肝脏包囊纤维化形成的病理学过程,本研究通过病理组织学、细胞化学和超微形态学的诊断方法,对羊感染细粒棘球蚴后肝脏的病理组织学变化,以及肝组织纤维化和包囊形成过程进行病理学观察。结果显示:原头蚴感染肝组织后病变沿感染组织的血管周围产生,相继出现肝细胞萎缩、变性、坏死;同时病变组织血管壁出现纤维组织分解、血管管壁出芽,增生出的纤维组织伸向周围炎症区域,血管管腔及炎区大量嗜酸性粒细胞浸润,增生的胶原纤维沿残存的肝细胞周围围绕病变组织,最终形成包囊壁。该研究结果为进一步阐释细粒棘球蚴感染羊肝脏包囊纤维化形成的病理机制奠定了坚实的基础。     

Congenital lipoblastoma in a neonate calf: first report in veterinary literature

Lipoblastoma is a rare benign tumour of adipose tissue that usually occurs in young children. Although around 100 cases of lipoblastoma have been reported in humans, no report has been previously described in animals. This case report describes the clinical and histopathological findings of a congenital lipoblastoma located at the tail base of a 2-day-old male calf. Grossly, the mass was pink with a partly ulcerated and haemorrhagic surface. The calf showed no neurological deficits. Radiographs of the vertebral column revealed no abnormalities, and surgical excision was performed. Histopathology of the mass was consistent with lipoblastoma. The calf was followed up for 13 months and found to be healthy with no signs of recurrence. To the best of the authors' knowledge, this is the first case of lipoblastoma in the veterinary literature. Lipoblastoma should thus be considered in the differential diagnosis list for congenital adipose tissue tumours in animals.