Geographic distribution and environmental risk factors of lymphoma in dogs under primary-care in the UK.

OBJECTIVES: To integrate external data sources with VetCompass postcode data to explore the spatial distribution and examine potential associations with environmental risk factors in dogs diagnosed with lymphoma at primary care veterinary practices. MATERIALS AND METHODS: Cases of lymphoma were identified from electronic patient records of 455,553 dogs under primary veterinary care during 2013 in the UK. Cases were defined as either laboratory-confirmed or non-laboratory-confirmed. Disease maps at the postcode-district level were used to define the geographic distribution of lymphoma incidence and spatial clustering was explored. Environmental risk factors from external data sources were transferred to a compatible format and logistic regression modelling was used to examine associations between environmental herbicide, fungicide and radon concentrations with lymphoma. RESULTS: From the denominator population of 455,553 dogs, 279 lymphoma cases (187 with laboratory confirmation and 93 without) were identified. Heterogeneous geographic variation was observed with weak evidence of clustering around London and the south-west of England. Herbicide and fungicide exposures were weakly associated with a diagnosis of lymphoma in the univariable analysis. After accounting for the age at diagnosis and breed in the multivariable analysis, herbicide exposure was associated with a diagnosis of lymphoma. CLINICAL SIGNIFICANCE: The heterogeneous distribution of lymphoma in UK dogs provides further evidence for geographic variation of lymphoma, perhaps in part associated with underlying environmental risk factors. The results suggest an association between environmental herbicide and canine lymphoma.

Significant advances in veterinary oncology - 60 years on.

The first edition of the Journal of Small Animal Practice published in February 1960, contained a paper entitled "Recent Treatments of Malignant Neoplasia" by Dr Larry Owen. Now we have reached the 60th anniversary of JSAP, that article provides a baseline from which to review subsequent advances in veterinary oncology, which now includes worldwide networks that have resulted in veterinary oncology becoming the multidisciplinary speciality that it is today. There certainly have been many advances in understanding of the pathology and epidemiology of animal cancers and in methods of diagnosis and treatment. However, the subject has become so large and diverse that not all aspects can be covered in detail here. It should also be acknowledged that there are still many gaps in knowledge in this field and that, because of a lack of randomised clinical trials, the evidence base for what is often regarded as "standard of care" is weak.

The future of radiotherapy in small animals - should the fractions be coarse or fine?

Radiation therapy has been used to treat animal cancers for more than 100 years. Clinical experiences and experimental results have been widely published and provide a basis for the recognition of radiation therapy as an integral component of multimodal cancer management in veterinary oncology. As the expectations of pet owners and the demand for treatment of companion animals with cancer have increased, veterinary oncology itself has undergone dramatic advances in the past several decades both in terms of improved diagnostics and treatments, including increased accessibility of radiation therapy. Synchronous with development of the specialism of veterinary radiation oncology, confusion and controversy have arisen with regard to distinguishing between different types of radiotherapy and methods of treatment delivery. Importantly, the confusion extends beyond semantics, and includes opinionated debate about defining which forms of therapy (if any at all) are optimal for a given patient. This exemplifies how, despite marks of maturity including age and a robust publication history, the field of veterinary radiation oncology is in some ways still in its infancy. The purpose of this article is to review the evidence base for daily (fine) fractionation versus weekly (coarse) hypofractionation in veterinary oncology, using selected tumour types as examples.

Temporal and spatial heterogeneity in milk and immune-related gene expression during mammary gland involution in dairy cows.

The aim of this study was to investigate heterogeneity in tissue morphology, milk protein and immune-related gene expression, and apoptosis of epithelial cells in the lactating and involuting mammary glands of the dairy cow. Mammary tissue from different regions of the gland (alveolar, cisternal, and peripheral) was collected postmortem from nonpregnant, pasture-fed, Holstein-Friesian primiparous cows in mid-lactation that were killed at different time points postmilking: 0, 6, 12, 18, 24, 36, and 72 h (n = 6 per time point). The CSN1NS1 and LALBA mRNA was decreased in alveolar, cisternal, and peripheral tissue by 12 to 36 h postmilking. In contrast, lactoferrin (LF) and mammary serum amyloid A3 (M-SAA3) mRNA was increased in these regions by 36 to 72 h. During lactation, more variability was present in gene expression in alveolar tissue between cows and between quarters within a cow, than within quarters. Histological analysis indicated the alveolar tissue from lactating cows was mostly uniform in structure; however, in situ hybridization indicated that although most of the alveolar tissue expressed milk proteins, the level of expression varied within and between alveoli. This heterogeneity became more pronounced with involution and with increasing regions of alveoli expressing lactoferrin, indicating that alveoli enter involution asynchronously. The peripheral and cisternal tissue had more variability in gene expression between cows compared with the alveolar tissue. The M-SAA3 signal was more intense in the cisternal tissue and less intense than the peripheral compartment compared with LF particularly in the earlier time points. In addition, between cows within the later time points, differences were observed in tissue morphology, the levels of milk protein and immune-related gene expression, and phosphorylated signal transducer and activator of transcription (STAT) 5-P and STAT3-P proteins, and degree of apoptosis, indicating that involution of the mammary gland occurs at different rates between cows. Understanding the mechanisms initiating the process of involution of the mammary gland provides an opportunity for enhancing milk production of the dairy cow.

Glycolysis inhibition improves photodynamic therapy response rates for equine sarcoids.

Photodynamic therapy (PDT) holds great promise in treating veterinary and human dermatological neoplasms, including equine sarcoids, but is currently hindered by the amount of photosensitiser and light that can be delivered to lesions thicker than around 2 mm, and by the intrinsic antioxidant defences of tumour cells. We have developed a new PDT technique that combines an efficient transdermal penetration enhancer solution, for topical delivery of 5-aminolevulinic acid (ALA) photosensitiser, with acute topical post-PDT application of the glycolysis inhibitor lonidamine. We show that the new PDT combination treatment selectively kills sarcoid cells in vitro, with repeated rounds of treatment increasing sarcoid sensitisation to PDT. In vivo, ALA PDT followed by 600 µM lonidamine substantially improves treatment outcomes for occult, verrucous, nodular and fibroblastic sarcoids after 1 month (93% treatment response in 27 sarcoids), compared with PDT using only ALA (14% treatment response in 7 sarcoids).

A retrospective review of treatment and response of high-risk mast cell tumours in dogs.

This retrospective case series evaluates survival outcome of 94 dogs with high metastatic risk mast cell tumours (MCT). Patients were treated with a cytotoxic chemotherapy protocol or the tyrosine kinase inhibitor masitinib, in the presence of gross disease or as an adjunct to surgical resection of the primary tumour. In patients presenting with metastatic disease, surgical resection of the primary tumour with adjunctive therapy with any chemotherapy incurred a significant survival advantage [median survival time (MST): 278 days] compared to patients receiving chemotherapy without surgical excision of the primary tumour (MST: 91 days, P < 0.0001). Patients with a surgically excised Patnaik grade II tumour and high Ki-67 in the absence of metastatic disease treated with vinblastine and prednisolone showed a significantly longer survival (MST: 1946 days) than those treated with masitinib (MST: 369 days, P = 0.0037). Further prospective case-controlled clinical trials of high-risk MCTs are required to make precise evidence-based treatment decisions for individual patients.

Diagnostic accuracy of three biopsy techniques in 117 dogs with intra-nasal neoplasia.

OBJECTIVES: To determine if nasal biopsies taken at rhinoscopy are more accurate for diagnosing neoplasia than biopsies taken blindly or using advanced imaging for guidance. METHODS: A retrospective study of 117 dogs with nasal mass lesions that were divided into three groups according to the method of nasal biopsy collection; advanced imaging-guided, rhinoscopy-guided and blind biopsy. Signalment, imaging and rhinoscopic findings, and histopathological diagnosis were compared between groups. The proportion of first attempt biopsies confirming neoplasia were determined for each group. RESULTS: There were no statistically significant differences in the proportion of biopsies that confirmed neoplasia obtained via advanced imaging-guided, rhinoscopy-guided or blind biopsy techniques. CLINICAL SIGNIFICANCE: In dogs with a high index of suspicion of nasal neoplasia, blind biopsy may be as diagnostic as rhinoscopy-guided biopsy. Repeated biopsies are frequently required for definitive diagnosis.

An immunohistochemical study of the expression of the hypoxia markers Glut-1 and Ca-IX in canine sarcomas.

Tumor hypoxia has been associated with increased malignancy, likelihood of metastasis, and increased resistance to radiotherapy and chemotherapy in human medicine. Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor that is induced by tumor hypoxia and regulates the pathways involved in cellular response and adaptation to the hostile tumor microenvironment. HIF-1 induces transcription of different proteins, including Ca-IX and Glut-1, which are considered endogenous markers of chronic hypoxia in solid tumors in humans. In this study, sections from 40 canine sarcomas (20 histiocytic sarcomas and 20 low-grade soft-tissue sarcomas) were immunostained for these markers. Expression of Glut-1 was scored based on percentage of positive staining cells (0 = <1%; 1 = 1%-50%; 2 = >50%) and intensity of cellular staining (1 = weak; 2 = strong); Ca-IX was scored based on percentage of positive cells (0 = <1%; 1 = 1%-30%; 2 = >30%). Intratumoral microvessel density was measured using CD31 to assess intratumoral neoangiogenesis. Histiocytic sarcomas showed statistically significant higher Glut-1 immunoreactivity and angiogenesis than did low-grade soft-tissue sarcomas. Intratumoral microvessel density in histiocytic sarcomas was positively associated with Glut-1 immunoreactivity score. These findings suggest a potential role of hypoxia in the biology of these tumors and may provide a base for investigation of the potential prognostic use of these markers in naturally occurring canine tumors.

Prognostic histopathological and molecular markers in feline mammary neoplasia.

Feline mammary tumours comprise approximately 11% of feline non-integumentary neoplasms, are more commonly malignant than benign, and carry a poor prognosis attributable to a high probability of local recurrence and metastasis. This review discusses histopathological and molecular markers that could aid in prognostic discrimination, and draws comparisons with studies examining prognostic markers in breast cancer. Tumour grade and mitotic index correlate with survival data and could be useful for prognostication. Although assessment of Ki67 expression might have prognostic potential, further studies are required to corroborate the correlation between expression and clinical outcome. Additional molecular markers that have been investigated for prognostic potential can be grouped according to the 'hallmarks of cancer'. Many studies utilise 'surrogate markers' of clinical outcome, such as correlation with histological grade, to assess the prognostic value of molecular markers, and further investigation is therefore necessary before reaching firm conclusions regarding the prognostic value of some markers. Feline mammary tumours have been proposed as spontaneous models of breast cancer but might only be suitable models for certain molecular sub-types. Compared to humans, cats tend to have a high percentage of mammary tumours which are oestrogen receptor-negative and they might therefore be suitable models for late stage oestrogen receptor-negative breast cancer. The basal-like properties of feline mammary carcinomas offer another avenue for future research in this field of comparative oncology.

COX-2 expression and outcome in canine nasal carcinomas treated with hypofractionated radiotherapy.

The expression of cyclooxygenase isoform 2 (COX-2) in canine nasal carcinomas has been well documented. COX-2 expression has proven to be a prognostic factor in several human tumours. The aims of this study were to assess the correlation between immunohistochemical COX-2 expression and prognosis using rhinoscopic biopsies from 42 dogs with nasal carcinomas treated with hypofractionated radiotherapy, and to establish a replicable COX-2 scoring system. Ninety per cent of sections evaluated were COX-2 positive with a mean score of 6.6 (median 8.0; range 0-12). Neither COX-2 expression nor tumour type had a significant correlation with survival. There are likely to be many as yet unidentified variants which contribute to length of survival in dogs with nasal carcinomas. Immunohistochemical COX-2 expression appears unlikely to be of prognostic significance for canine nasal carcinoma.

The clinical presentation and histopathologic-immunohistochemical classification of histiocytic sarcomas in the Flat Coated Retriever.

The Flat Coated Retriever is a breed at risk of development of histiocytic sarcoma (HS), but in contrast to the disseminated form of disease recognized in the Bernese Mountain Dog, most reports of HS in Flat Coated Retrievers describe a localized lesion affecting the musculature or fascia of limbs. The purpose of this study was to review data and material received though an ongoing Flat Coated Retriever tumor survey to better define the presentation of HS in the breed and to determine the utility of subclassification of tumors arising at different sites by histology and immunohistologic phenotyping. Data on 180 dogs bearing HS-like tumors were available for review, which showed that although the majority (101 lesions, 57%) were primary limb lesions, 47 dogs (26%) had visceral, mainly splenic lesions with no peripheral primary tumor. A detailed histologic and immunohistologic review of 20 limb tumors and 20 splenic tumors showed that 2 distinct phenotypic subtypes could be identified: a histiocytic subtype, most prevalent in the splenic tumors, and a histiocytic-spindle-pleomorphic subtype, mainly seen in the limb tumors. Despite their variable morphology, all tumors expressed major histocompatibility complex class II and the leukocyte antigen CD18, but only those tumors in the spleen consistently expressed CD11d. The majority of tumors also contained a mild to moderate infiltrate of T lymphocytes.

Prognostic significance of specific magnetic resonance imaging features in canine nasal tumours treated by radiotherapy.

OBJECTIVES: To investigate the prognostic significance of the magnetic resonance (MR) findings of meningeal hyperintensity of the olfactory bulbs and tumour extension into the caudal nasal recess (CNR) in dogs with nasal tumours treated by radiotherapy. METHODS: MR images of 41 dogs with nasal tumours treated with radiotherapy were reviewed. The occurrence of neurological signs and survival of patients with and without meningeal hyperintensity of the olfactory bulbs and tumour extension into the CNR were analysed together with possible confounding factors including intracranial extension and patient age. RESULTS: There was no significant association between the presence of meningeal hyperintensity or CNR involvement and the occurrence of neurological signs. Although there was a tendency towards shorter survival in dogs with tumour extension into the CNR, multivariable analysis showed no significant difference in survival between dogs with/without CNR involvement, meningeal hyperintensity or intracranial tumour extension (P=0.12, 0.50 and 0.57, respectively). CLINICAL SIGNIFICANCE: In dogs with nasal tumours treated with radiotherapy, tumour extension into the cranium is not necessarily associated with shorter survival in patients without neurological signs at time of diagnosis. Although a definite influence of CNR involvement on case outcome could not be demonstrated, studies with a larger population are warranted.

Feline extranodal lymphoma: response to chemotherapy and survival in 110 cats.

OBJECTIVE: To determine response to treatment, survival and prognostic factors for feline extranodal lymphoma in the UK. METHODS: Records of cats diagnosed with lymphoma of extranodal sites at seven referral centres were reviewed and information on signalment, tumour location, prior treatment and chemotherapy protocol recorded. Factors influencing response to treatment and survival were assessed. RESULTS: One hundred and forty-nine cases met inclusion criteria. Sixty-nine cats had nasal lymphoma, 35 renal, 15 central nervous system, 11 laryngeal and 19 miscellaneous locations. Sixty-six cats received cyclophosphamide, vincristine, prednisolone, 25 Wisconsin-Madison doxorubicin-containing multi-agent protocol, 10 prednisolone alone and nine other combinations. The response rate for the 110 treated cats was 85.5 per cent. Of cyclophosphamide, vincristine, prednisolone treated cats 72.7 per cent achieved complete remission, median survival 239 days. Sixty-four per cent of Wisconsin-Madison treated cats achieved complete remission, median survival 563 days. Cats with nasal lymphoma achieving complete remission had the longest survival (749 days) and cats with central nervous system lymphoma the shortest (70 days). If complete remission was achieved, prior treatment with corticosteroids significantly reduced survival time. CLINICAL SIGNIFICANCE: Cats with extranodal lymphoma respond to chemotherapy and achieve survival times comparable to other locations. Corticosteroid pretreatment reduced survival time in cats achieving complete remission.

Exogenous insulin treatment after hypofractionated radiotherapy in cats with diabetes mellitus and acromegaly.

BACKGROUND: The optimal treatment for feline acromegaly has yet to be established. Surgical and medical therapies are minimally effective although radiotherapy might have greater efficacy. The purpose of this study was to review the response and outcome of cats with acromegaly and insulin-resistant diabetes mellitus (DM) to radiotherapy. HYPOTHESES: That radiotherapy improves glycemic control in cats with acromegaly and that improved glycemic control is due to remission of clinical acromegaly; demonstrated by a fall in serum insulin-like growth factor-1 (IGF-1) concentrations. ANIMALS: Fourteen cats with naturally occurring acromegaly. METHODS: Retrospective case review; records of all cats treated for acromegaly with radiotherapy were reviewed from 1997 to 2008. Cats were selected on the basis of compatible clinical signs, laboratory features, and diagnostic imaging findings. Fourteen cats received radiotherapy, delivered in 10 fractions, 3 times a week to a total dose of 3,700 cGy. RESULTS: Thirteen of 14 cats had improved diabetic control after radiotherapy. These improvements were sustained for up to 60 months. DM progressed in 2 cats and 1 did not respond. Seven cats responded before the final treatment. Ten cats were euthanized, 1 as a consequence of radiotherapy. In 8 cats in which IGF-1 was measured after treatment, changes in its concentration did not reflect the clinical improvement in glycemic control. CONCLUSIONS AND CLINICAL IMPORTANCE: Radiotherapy represents an effective treatment for cats with insulin-resistant DM resulting from acromegaly. IGF-1 concentration after treatment does not provide a suitable method by which remission from either acromegaly or insulin-resistant DM may be assessed.

Photodynamic therapy of superficial nasal planum squamous cell carcinomas in cats: 55 cases.

BACKGROUND: Squamous cell carcinomas (SCCs) are common skin tumors in cats. We investigated photodynamic therapy (PDT) using the photosensitizing agent 5-aminolaevulinic acid (5-ALA) topically and a high-intensity red light source. HYPOTHESIS: PDT is a safe and effective treatment for feline SCCs. ANIMALS: Fifty-five client-owned cats with superficial nasal planum SCCs. METHODS: Prospective, uncontrolled clinical trial. PDT was performed using topical 5-ALA and light of peak wavelength 635 nm. Adverse effects, response, and tumor control were evaluated. RESULTS: 53/55 (96%) cats responded to therapy, and there was a complete response in 47/55 (85%). Six cats (11%) had a partial response. Of the 47 cats with complete response to a single treatment, 24 recurred (51%), with a median time to recurrence of 157 days (95% confidence interval, 109-205 days). Repeat PDT was performed in 22 cats, and at a median follow-up of 1,146 days, 23 (45%) cats were alive and disease free, 17 (33%) had to be euthanized due to tumor recurrence, and 11 (22%) were euthanized for other reasons. Only transient mild local adverse effects were observed after treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: PDT using 5-ALA and a red light source was safe, well tolerated, and effective in the treatment of superficial nasal planum SCCs of cats and offers an alternative to conventional therapy. Although initial response rates were high, this treatment did not lead to a durable remission or cure in all cases.

Retrospective review of 50 canine nasal tumours evaluated by low-field magnetic resonance imaging.

OBJECTIVES: Low-field magnetic resonance imaging machines are being used more often in veterinary practice for the investigation of sinonasal disease. The aim of this retrospective study was to describe and characterise the low-field magnetic resonance imaging features of nasal tumours in dogs. METHODS: The Queen's Veterinary School Hospital magnetic resonance imaging database (2001-2005) was searched for dogs with a magnetic resonance imaging diagnosis of a nasal tumour. Fifty cases with histological diagnosis of nasal tumour were found. The appearance and extent of the nasal tumour as well as the involvement of adjacent anatomic structures were examined against a checklist. RESULTS: The most common magnetic resonance imaging findings were as follows. (1) Soft tissue mass replacing the destroyed nasal conchae and/or ethmoturbinates (98 per cent of cases). (2) Nasal septum destruction (68 per cent of cases). (3) Retained secretions with or without mass caudally in frontal sinuses (62 per cent of cases). (4) Nasal/frontal bone destruction (52 per cent of cases). Low-field magnetic resonance imaging allowed differentiation of tumour tissue from retained secretions or necrotic tissue. Magnetic resonance imaging was invaluable in assessing the extension of the tumour into the maxillary recesses, caudal recesses, nasopharynx, adjacent bones and cranial cavity. The tumour often extended caudally into the frontal sinuses, nasopharynx and perhaps most importantly into the caudal recesses. Tumour extension into the cranial cavity was not common (16 per cent), and only three of these cases showed neurological signs. However, 54 per cent of cases showed focal meningeal (dural) hyperintensity, although the significance of this is unclear. A significant difference (P<0.05) in tumour signal intensity between the sarcomas and carcinomas was found. CLINICAL SIGNIFICANCE: The use of a low-field magnetic resonance imaging technique is excellent for the diagnosis and determination of extent of sinonasal tumours.

Advances in the diagnosis and management of cutaneous mast cell tumours in dogs.

Mast cell tumours are one of the most common tumours of the canine skin and have a reputation for being difficult to manage because of their variable clinical presentation, behaviour and response to treatment. This review of recent literature on canine mast cell tumours suggests that the majority of such tumours may not be as bad as their reputation suggests. Most grade I and grade II tumours can be managed successfully by good surgery. Recent literature also calls into question the utility of clinical staging systems and the value of assessing surgical margins for prognosis and highlights the paucity of well-conducted, case-controlled clinical trials in assessing the efficacy of medical management of high-risk tumours. In terms of more basic research, recent studies have implicated the stem cell factor receptor KIT as having a role in the aetiology of canine mast cell tumours and there appears to be an association between c-kit mutation and higher grade of tumour. This may offer a possible target for new therapeutic approaches.

Chromosome rearrangements in canine fibrosarcomas.

We have previously reported the use of six- and seven-color paint sets in the analysis of canine soft tissue sarcomas. Here we combine this technique with flow sorting of translocation chromosomes, reverse painting, and polymerase chain reaction (PCR) analysis of the gene content of the reverse paint in order to provide a more detailed analysis of cytogenetic abnormalities in canine tumors. We examine two fibrosarcomas, both from female Labrador retrievers, and show abnormalities in chromosomes 11 and 30 in both cases. Evidence of involvement of TGFBR1 is presented for one tumor.

Immune-mediated haemolytic anaemia associated with a sarcoma in a flat-coated retriever.

A seven-year-old flat-coated retriever presented with a history of lethargy, dyspnoea and inappetence of several days' duration. Clinical examination revealed pale mucous membranes and tachypnoea, and haematology demonstrated marked autoagglutination. Thoracic radiographs revealed an increased opacity in the perihilar region. The owners declined further evaluation and the dog was treated symptomatically with immunosuppressive doses of prednisolone and azathioprine. The dog's demeanour improved, although it was eventually euthanased seven weeks later because of dysphagia and worsening dyspnoea. Postmortem examination revealed a widespread, poorly differentiated sarcoma involving the lungs, pericardium, thoracic lymph nodes and spleen. Immune-mediated haemolytic anaemia is a well recognised condition in dogs and is occasionally associated with neoplastic conditions. This is the first case report to describe immune-mediated haemolytic anaemia associated with a diffuse, poorly differentiated sarcoma.

Owners' assessments of their dog's quality of life during palliative chemotherapy for lymphoma.

The owners of 25 dogs with multicentric lymphoma which were treated by multidrug chemotherapy were interviewed to assess the quality of their pet's life during treatment. Sixty-eight per cent of the owners considered their dog's quality of life to be the same as before the lymphoma occurred, and the remaining 32 per cent felt that their pet's quality of life on chemotherapy treatment was acceptable but poorer than before the lymphoma occurred. Treatment complications were reported by 52 per cent of owners. Ninety-two per cent of owners had no regrets about treating their dog with multidrug chemotherapy. The results suggest that, while many dogs developed treatment complications during multidrug chemotherapy, most owners felt that the quality of life of their pets was good during treatment and had no regrets about opting for palliative chemotherapy.