Features and prognostic impact of distant metastases in 45 dogs with de novo stage IV cutaneous mast cell tumours: A prospective study

BACKGROUND: Distant metastases in dogs with cutaneous mast cell tumors (cMCT) are rare and incurable. The aims of this prospective study were to clarify the clinico-pathological features of stage IV cMCTs and to identify possible prognostic factors for progression-free interval (PFI) and survival time (ST). MATERIAL AND METHODS: Dogs were eligible for recruitment if they had a previously untreated, histologically confirmed cMCT and if they underwent complete staging demonstrating stage IV disease. Dogs were uniformly followed-up, whereas treatment was not standardized and included no therapy, surgery, radiation therapy, chemotherapy, tyrosine-kinase inhibitors or a combination of these. RESULTS: 45 dogs with stage IV cMCT were enrolled. All dogs had distant metastatic disease, and 41 (91.1%) dogs had also metastasis in the regional lymph node. Histopathological grade and mutational status greatly varied among dogs. Median ST was 110 days. Notably, PFI and ST were independent of well-known prognostic factors, including anatomic site, histological grade, and mutational status. Conversely, tumor diameter >3\u2009cm, more than 2 metastatic sites, bone marrow infiltration, and lack of tumor control at the primary site were confirmed to be negative prognostic factors by multivariate analysis. CONCLUSION: Currently, there is no satisfactory treatment for stage IV cMCT. Asymptomatic dogs with tumor diameter <3\u2009cm and a low tumor burden, without bone marrow infiltration may be candidates for multimodal treatment. Stage IV dogs without lymph node metastasis may enjoy a surprisingly prolonged survival. The achievement of local tumor control seems to predict a better outcome in dogs with stage IV cMCT

Immunohistochemical Expression of FXR1 in Canine Normal Tissues and Melanomas

Fragile X mental retardation-related protein 1 (FXR1) is a cytoplasmic RNA-binding protein highly conserved among vertebrates. It has been studied for its role in muscle development, inflammation, and tumorigenesis, being related, for example, to metastasizing behavior in human and canine uveal melanoma. Anti-FXR1 antibodies have never been validated in the canine species. To investigate FXR1 expression in canine melanocytic tumors, the present study tested two commercially available polyclonal anti-human FXR1 antibodies, raised in goat and rabbit, respectively. The cross-reactivity of the anti-FXR1 antibodies was assessed by Western blot analysis, and the protein was localized by IHC in a set of normal canine tissues and in canine melanocytic tumors (10 uveal and 10 oral). Western blot results demonstrated that the antibody raised in rabbit specifically recognized the canine FXR1, while the antibody raised in goat did not cross-react with this canine protein. FXR1 protein was immunodetected using rabbit anti-FXR1 antibody, in canine normal tissues with different levels of intensity and distribution. It was also detected in 10/10 uveal and 9/10 oral melanocytic tumors. The present study validated for the first time the use of anti-FXR1 antibody in dogs and highlighted different FXR1 protein expression in canine melanocytic tumors, the significance of which is undergoing further investigations

Conformity and controversies in the diagnosis, staging and follow-up evaluation of canine nodal lymphoma: a systematic review of the last 15 years of published literature

Diagnostic methods used in the initial and post-treatment evaluation of canine lymphoma are heterogeneous and can vary within countries and institutions. Accurate reporting of clinical stage and response assessment is crucial in determining the treatment efficacy and predicting prognosis. This study comprises a systematic review of all available canine multicentric lymphoma studies published over 15 years. Data concerning diagnosis, clinical stage evaluation and response assessment procedures were extracted and compared. Sixty-three studies met the eligibility criteria. Fifty-five (87.3%) studies were non-randomized prospective or retrospective studies. The survey results also expose variations in diagnostic criteria and treatment response assessment in canine multicentric lymphoma. Variations in staging procedures performed and recorded led to an unquantifiable heterogeneity among patients in and between studies, making it difficult to compare treatment efficacies. Awareness of this inconsistency of procedure and reporting may help in the design of future clinical trials

Prognostic significance of peripheral blood and bone marrow infiltration in newly-diagnosed canine nodal marginal zone lymphoma

Canine nodal marginal zone lymphoma (nMZL) is infrequent and is typically diagnosed at an advanced disease stage. However, it is currently unknown whether different levels of peripheral blood (PB) and bone marrow (BM) infiltration may provide prognostic stratification in dogs with nMZL. The aims of the present prospective study were to assess the influence of PB and BM infiltration detected by flow cytometry (FC) on time to progression (TTP) and lymphoma-specific survival (LSS) in dogs with newly-diagnosed multicentric nMZL, and to establish a cut-off value of prognostic significance. Forty-five completely staged and treatment-na\ueff dogs with histologically-confirmed nMZL were enrolled. After staging, dogs received chemo-immunotherapy or chemotherapy. PB infiltration was significantly associated with TTP (p = 0.001): dogs with PB infiltration <30% had a median TTP of 186 days, whereas dogs with PB infiltration 6530% had a median TTP of 43 days. Additionally, vaccinated dogs had a significantly (p = 0.012) longer TTP (399 days) compared with dogs receiving chemotherapy only (211 days). BM infiltration was significantly associated with LSS (p < 0.001): dogs with BM infiltration <1% had a median LSS of 1403 days, those with BM infiltration 1\u201320% of 337 days, and those with BM infiltration 6520% of 188 days. Normal LDH levels and the administration of chemo-immunotherapy also significantly improved LSS (560 vs 211 days, and 399 vs 211 days, respectively; p < 0.001). PB and BM flow cytometric evaluation is an integral part of staging work-up in dogs with nMZL and has prognostic relevance

Treatment Effects and Risk Factors Evaluation in Longitudinal Studies : A Statistical Help for Data Analysis

This paper was inspired by the experience of the Authors research group composed by oncologist veterinarians and a biostatistician to evaluate treatments and prognostic factors with the aim to help veterinarians involved in longitudinal studies into evaluating and writing prognostic results. Longitudinal studies are commonly analysed by techniques for survival data, taking into account for the time elapsed from the beginning of observation and the occurrence of an event related to treatment effect or disease course. The presence of incomplete follow-up information for some subjects requires specific descriptive and inferential statistical methods. Two literature datasets were analysed to show statistical models implementation techniques and to discuss statistical issues: I) A multicentre clinical trial on remission maintenance of children with acute Lymphoblastic leukaemia and II) A randomized clinical trial on advanced inoperable lung cancer. Data sets concerned studies on \u201chumans\u201d, nevertheless the peculiar data structure allowed to discuss some aspects which are common to survival analysis studies, regardless on subject\u2019s characteristics. Log-rank test was used to compare survival curves for treatments and the relationship between Log-Rank test and univariate Cox model results was explained. As the evaluation of prognostic impact cannot be based only on p-values, the strength of the association between treatments and prognosis was estimated to take into account for the clinical relevance of results. On the second data set, beside of treatment, other clinical variables were available and a multivariate Cox model was applied. Model implementation was discussed concerning the coding of categorical variables and the relationship between continuous variables and model response. Suggested rules for the maximum number of covariates to be included in order to obtain reliable results were cited. Finally, the predictive ability of the model was discussed based on a measure of the area under ROC curve specific for survival data

Prognostic role of non-neoplastic lymphocytes in lymph node aspirates from dogs with diffuse large B-cell lymphoma treated with chemo-immunotherapy

Dogs with Diffuse Large B-Cell Lymphoma (DLBCL) benefit from the addition of active immunotherapy to traditional chemotherapy. We hypothesized that immune cells within neoplastic lymph nodes (LNs) may play a role in the tumor pathobiology and treatment response. The present study describes the composition and prognostic role of non-neoplastic lymphocytes in LNs of 59 dogs with treatment-naive DLBCL receiving chemo-immunotherapy. The percentage of small non-neoplastic cells and of CD5+, CD21+, CD4+ and CD8+ small cells was recorded via flow cytometry. CD4+/CD8+ and CD5+/large CD21+ cell ratios were calculated. The likelihood of progression significantly diminished with increasing percentage of small cells, CD5+ and CD8+ small cells, and CD5+/large CD21+ cell ratio, with decreasing CD4+/CD8+ ratio and in non-anemic dogs. Active immunotherapy is more effective in dogs with higher percentage of non-neoplastic lymphocytes at diagnosis. We lay the ground for future studies assessing the role of the immune system in the pathobiology of canine DLBCL

Proximal mandibular nerve block using eletrolocation, for rostral mandibulectomy in a geriatric dog

Abstract \u2014 We describe a case of proximal mandibular nerve block with ropivacaine, using electrolocation, for perioperative pain management in a geriatric dog undergoing rostral mandibulectomy. The patient did not require intraoperative analgesia or analgesic supplementation for 8 h after the end of the surger

Effects of pre-analytical variables on flow cytometric diagnosis of canine lymphoma : A 3 retrospective study (2009-2015)

Flow cytometry (FC) is increasingly being used for immunophenotyping and staging of canine lymphoma. The aim of this retrospective study was to assess pre-analytical variables that might influence the diagnostic utility of FC of lymph node (LN) fine needle aspirate (FNA) specimens from dogs with lymphoproliferative diseases. The study included 987 cases with LN FNA specimens sent for immunophenotyping that were submitted to a diagnostic laboratory in Italy from 2009 to 2015. Cases were grouped into 'diagnostic' and 'non-diagnostic'. Pre-analytical factors analysed by univariate and multivariate analyses were animal-related factors (breed, age, sex, size), operator-related factors (year, season, shipping method, submitting veterinarian) and sample-related factors (type of sample material, cellular concentration, cytological smears, artefacts). The submitting veterinarian, sample material, sample cellularity and artefacts affected the likelihood of having a diagnostic sample. The availability of specimens from different sites and of cytological smears increased the odds of obtaining a diagnostic result. Major artefacts affecting diagnostic utility included poor cellularity and the presence of dead cells. Flow cytometry on LN FNA samples yielded conclusive results in more than 90% of cases with adequate sample quality and sampling conditions

PRELIMINARY STUDY OF WATER INJECTION ON THE COMBUSTION AND EMISSIONS CHARACTERISTICS IN A HCCI ETHANOL ENGINE

Our dependence on fossil fuels coupled with concerns about harmful emissions have motivated researchers to look for renewable fuels that have clean combustion and for advanced combustion modes. In this context, homogeneous charge compression ignition (HCCI) is an emerging technology which offers an alternative to conventional spark ignition and compression ignition engines and can operate on renewable fuels. Low temperature combustion, which can result in low NOx emissions with high indicated efficiency, is the more important characteristic of this combustion mode. It’s main problem is the combustion timing control due to lack of direct ignition control, once HCCI flame initiation is based on charge thermal state. Thus, controlled auto-ignition (CAI) combustion mode has been proposed. Several methods were proposed for combustion phasing control, between them, the injection of water in the intake manifold. This work investigated the influence of water injection in the intake runner of an ethanol HCCI cylinder from a converted three-cylinder diesel generator set, in which two cylinders operated on conventional diesel combustion and one diesel cylinder provided recycled exhaust gas for the one cylinder running on ethanol HCCI combustion. The water injection was used to control the CA50 combustion parameter. The results show that water injection is an efficient strategy to control the combustion timing, since the reactivity of the mixture can be controlled. The results at 400 and 600 kPa of IMEP and 1800 rpm indicated a good combustion stability, high efficiency and low emissions characteristics. The highest indicated fuel conversion efficiency found was 36.9% for 600 kPa of IMEP and 8 CAD of CA50. However, for 200 kPa of IMEP the combustion was unstable, the indicated efficiency was deteriorated and indicted CO emissions was high