Analytical Validation of MINI-PET as Point-of-Care for Erythrocyte Sedimentation Rate Measure in Horses.

The erythrocyte sedimentation rate (ESR) is a widely used diagnostic assay in human medicine but nowadays poorly applied in veterinary medicine. This test measures the speed (millimeters per hour) at which red blood cells settle in a whole anticoagulated blood tube. In human medicine, high ESR values are associated with various disorders, including infections, rheumatoid arthritis, oncologic diseases, and other inflammatory conditions. The ESR can also be influenced by some factors such as age and gender. In veterinary medicine, the ESR with the Westergren manual method was almost forgotten over the years due to blood consumption and long turn-around time. The instrument MINI-PET, using a modified Westergren method, does not require blood consumption or release waste product and recently has been applied in canine medicine. The aims of the study in the horse were as follows: to establish the appropriate time to read the ESR with the Westergren reference method; to compare the MINI-PET ESR results with the reference technique; to assess the ESR reference intervals with MINI-PET; and to establish the ESR stability from collection at different time points by MINI-PET. Using 150 horses, we established 60 minutes as the appropriate time for ESR reading with the Westergren method. Moreover, ESR results obtained in 8 minutes with MINI-PET showed a good correlation with the Westergren ESR. Reference intervals (RIs) with MINI-PET were established in mm/h for the healthy horses (geldings 18.6-100.1; stallions, 13.8-55.7; and mares 1-73.7) according to the American Society of Veterinary Clinical Pathology guidelines. In addition, the ESR stability from the blood collection time was evaluated in the MINI-PET on 15 horses: at room temperature, ESR is stable up to 8 hours and at 4°C up to 24 hours. In conclusion, MINI-PET represents a rapid and reliable tool for measuring ESR in horses, offering a valid option to replace the traditional manual technique.

Utility and prognostic significance of leukocyte ratios in dogs with Primary Immune-Mediated Hemolytic Anemia.

Canine immune-mediated hemolytic anemia (IMHA) is a life-threatening condition that is commonly associated with neutrophilia and monocytosis. Leukocyte ratios have been found to have prognostic value in humans and animals affected by a range of inflammatory, infectious, and neoplastic disorders. We hypothesized that in primary IMHA, neutrophil to lymphocyte (NLR), neutrophil to monocyte (NMR), band neutrophil to segmented neutrophil (BNR) and monocyte to lymphocyte (MLR) ratios would be higher in dogs that did not survive to discharge. Medical records of dogs diagnosed with IMHA at two veterinary teaching hospitals were retrospectively reviewed. Twenty-three of the 72 included dogs did not survive to discharge. NLR, NMR, BNR and MLR ratios were compared between dogs that survived to discharge and dogs that died or were euthanized. None of the ratios were significantly different between survivors and non-survivors (P = 0.14-0.99). Area under the Receiver Operating Characteristic (ROC) curve for prediction of non-survival ranged from 0.5 (95% confidence interval 0.38-0.62) for MLR to 0.61 (0.49-0.72) for NMR and was not significantly different from 0.5 for any ratio (P = 0.29-0.99). After exclusion of 31 dogs that received one or both immunosuppressive medications and blood transfusion before presentation, the area under the ROC curve for prediction of survival was significantly different from 0.5 for MLR (0.78, P = 0.01) and NMR (0.78, P = 0.0002). This study suggests that lower MLR and higher NMR may predict poorer prognosis in untreated dogs with IMHA.

Multiple myeloma and primary erythrocytosis in a dog.

A 13-year-old spayed female mixed breed dog was referred for impaired ambulation, limb tremors, back pain, hypergammaglobulinemia on cellulose acetate electrophoresis, and mild proteinuria. Conventional radiology and magnetic resonance imaging (MRI) suggested multifocal neoplastic bone lesions. At the referral examination, lameness and bright red mucous membranes were observed. Severe erythrocytosis, a monoclonal peak in the ß-2 globulin detected by capillary zone electrophoresis, severe proteinuria, bone marrow infiltration of plasma cells, and low serum erythropoietin concentrations were reported. The final diagnosis was multiple myeloma associated with severe primary erythrocytosis. This presentation in a dog is interesting because the combination of both disorders is rare in humans and has not been reported in dogs. Key clinical message: Although rare, multiple myeloma and primary erythrocytosis can occur together in dogs.

Myélome multiple et érythrocytose primaire chez un chien. Une chienne de race mixte stérilisée âgée de 13 ans a été référée pour troubles de la marche, tremblements des membres, maux de dos, hypergammaglobulinémie à l'électrophorèse sur acétate de cellulose et protéinurie légère. La radiologie conventionnelle et l'imagerie par résonance magnétique (IRM) suggéraient des lésions osseuses néoplasiques multifocales. Lors de l'examen de référence, une boiterie et des muqueuses rouge vif ont été observées. Une érythrocytose sévère, un pic monoclonal de la globuline ß-2 détecté par électrophorèse capillaire, une protéinurie sévère, une infiltration de la moëlle osseuse par des plasmocytes et de faibles concentrations sériques d'érythropoïétine ont été rapportés. Le diagnostic final était un myélome multiple associé à une érythrocytose primaire sévère. Cette présentation chez un chien est intéressante car l'association des deux conditions est rare chez l'homme et n'a pas été rapportée chez le chien.Message clinique clé :Bien que rares, le myélome multiple et l'érythrocytose primaire peuvent survenir simultanément chez le chien.(Traduit par Dr Serge Messier).

A Retrospective Multicentric Study of Electrochemotherapy in the Treatment of Feline Nasal Planum Squamous Cell Carcinoma.

Feline squamous cell carcinoma (SCC) is currently treated with surgery, radiation therapy and electrochemotherapy (ECT). Both the efficacy and/or safety of ECT were evaluated as a sole therapy with bleomycin to treat feline nasal planum SCC (npSCC). Sixty-one cats were enrolled. Local treatment response was evaluated as complete remission (CR), partial remission (PR) or stable disease (SD). Recurrence rate (RR), disease-free interval (DFI) and progression free survival (PFS) were calculated. A six-point scale was used for ECT toxicity. The median tumor size was 1.5 cm. CR was achieved in 65.6% of cases, PR in 31.1% and SD in 3.3%. The overall response rate was 96.7%, RR was 22.5%, median DFI was 136 days, and median PFS was 65.5 days. ECT toxicity was ≤2 in 51% of cats. Tumor recurrence/progression (p = 0.014) and local treatment response (PR: p < 0.001; SD: p < 0.001) influenced survival time. Cats with toxicity >2 showed a higher probability of tumor recurrence/progression. Tumor-related death was higher in cats with PR (p < 0.001) and recurrence/progression (p = 0.002), in ECT treatment with 1 Hz (p = 0.035) and 1200 V/cm (p = 0.011) or 1300 V/cm (p = 0.016). Tumor size influenced local treatment response (p = 0.008) and toxicity (p < 0.001). ECT is an effective treatment for feline npSCCs and should be considered as the first-line procedure for low-stage tumors.

The guaiac-based fecal occult blood test in healthy dogs: Evaluation of the effects of diet, and the ability of the test to detect fecal occult blood.

BACKGROUND: The guaiac-based fecal occult blood test (gFOBt) has been used for human colorectal cancer screening. It can detect the fecal occult blood (FOB) in dogs after oral administration of 20 mg of hemoglobin/kg body weight of blood and is influenced by diet. OBJECTIVES: The aims were to evaluate the effect of diet and the ability of gFOBt to detect FOB in healthy dogs after oral administration of autologous blood. METHODS: Five healthy dogs were fed Purina Hypoallergenic (HA) and Gastrointestinal (EN) diets. Feces were tested with gFOBt before starting diets and at each defecation (hereafter referred to as fecal collection event) throughout the study period. Every 4 days, increased doses of autologous blood were administered orally. The whole blood of one dog was progressively diluted with a saline solution, and dilutions were directly tested with the gFOBt, until a negative result was found. RESULTS: Twelve of 185 (6.5%) gFOBt were found to be positive. No associations between diet and gFOBt positivity were found. A significant association was found between the fecal collection event and gFOBt positivity (P < .001) and between doses of blood and gFOBt positivity (P = .048). The lowest dilution ensuring all positive tests was 6.5 µgHgb /mL. CONCLUSIONS: The gFOBt was not influenced by either the HA or EN diets. The gFOBt positivity was associated with the dose of blood and the fecal collection event. However, caution is needed in the interpretation of results due to the lack of an association between gFOBt positivity and increasing doses of blood in the same dog.

Electrochemotherapy in treatment of canine oral non-tonsillar squamous cell carcinoma. A case series report.

Non-tonsillar squamous cell carcinoma (ntSCC) is a common and locally aggressive oral tumour in dogs. The treatments of choice are currently surgery and radiotherapy. Electrochemotherapy (ECT) is a local ablative anti-tumour technique using electric pulses to enhance the intracellular diffusion of cytotoxic drugs. The aim was to retrospectively evaluate the outcome of patients with oral ntSCC treated with ECT. Twelve dogs with ntSCC were retrospectively enrolled. ECT was combined with IV bleomycin (15 000 UI/m2 ) alone in 11 cases and post-surgery in 1. Parameters considered were: tumour site and size, electroporation parameters, response rate (complete remission [CR], partial remission [PR]), median survival time (MST), recurrence rate (RR), median disease-free interval (DFI) and treatment toxicity (6-point scale). Median tumour size was 1.65 cm (range 0.3-8.0 cm) and the response rate was 90.9% (10/11; 8 CR and 2 PR). Two dogs underwent a second ECT. MST for dogs dead with tumour (n = 2) was 110 days and for dogs dead without tumour (n = 3) was 831 days. Among five surviving dogs, one experienced tumour recurrence and four were in CR. Results from two dogs were analysed separately. Overall RR was 27.3%. DFI and MST for dogs with recurrence were 50 and 115 days, respectively. Treatment toxicity was very low. We noticed that all dogs with tumours smaller than 1-2 cm achieved CR without recurrence suggesting a favourable prognosis when using ECT. ECT for canine ntSCC could be considered a valid treatment option especially for smaller tumours, but a larger caseload would be needed to confirm this statement.

Bleeding diathesis in canine multiple myeloma and prognostic implications: A cohort study in 156 dogs.

Multiple myeloma (MM) is a tumor of plasma cells representing approximately 1% of all canine tumors. Clinical evident bleeding is often referred to as the main finding. The aim of the study was to evaluate the occurrence of clinical bleedings in dogs with MM and its prognostic implications compared to a population of dogs not affected by MM. Two groups of dogs (# 78 each) individually matched for breed, age and gender were considered. Group-1 (exposed) was affected by MM and group-2 (unexposed) was affected by other diseases. They were compared for bleeding and mortality at 90 days after diagnosis (relative risk, RR; attributable risk, AR). Among group-1, bleeding patients (B) were compared with non-bleeding patients (NB) in terms of mortality at 90 days (RR, AR). Incident cases of MM were 78/57,694 (0.13%). Signs of bleeding up to 30 days before the referral presentation were found in 33 (42.3%) group-1 dogs in comparison to 6 (7.7%) group-2 dogs (RR, 5.50, CI 95% 2.55-12.3, p = 0.0001; AR, 0.34, CI 95% 0.22-0.47, p = 0.0001). Epistaxis was the most frequent sign of bleeding recorded. Nineteen dogs from group-1 (24.3%) and eight from group-2 (10.2%) were non-survivors (RR = 2.37, CI 95% 1.14-5.06, p = 0.01; AR = 0.14, CI 95% 0.02-0.26, p = 0.01). Among the group-1, the B dogs, 4/33 (12.1%) were non-survivors, while 15/45 NB dogs (33.3%) were non-survivors (RR = 2.75, CI 95% 1.08-7.44, p = 0.03; AR = 0.21, CI 95% 0.20-0.38, p = 0.03). Epistaxis at diagnosis was frequent in MM dogs, and signs of bleeding were associated with a more favorable 90-day prognosis.

Epidemiology of Breed-Related Mast Cell Tumour Occurrence and Prognostic Significance of Clinical Features in a Defined Population of Dogs in West-Central Italy.

Canine mast cell tumours (MCTs) present a wide variety of challenging clinical behaviours in terms of predicting the prognosis and choosing appropriate treatment. This study investigated the frequency, risk, and prognostic factors of MCTs in dogs admitted to a single veterinary teaching hospital (VTH). Breed, age, sex, and sexual status in ninety-eight dogs with MCTs (MCT-group) were compared with a control group of 13,077 dogs (VTH-group) obtained from the VTH clinical database from January 2010 to January 2016. Within the MCT-group, signalment, location, size, mass number, ulceration, histopathological grading, presence of lymph node, or distant metastases were compared with each other and with the outcome. Boxers (OR 7.2), American Pit Bull Terriers (OR 5.4), French Bulldogs (OR 4.4) and Labrador Retrievers (OR 2.6) were overrepresented. The MCT-group was significantly older than the VTH-group (p < 0.0001). In comparison with the VTH group, in the MCT-group neutered dogs (OR 2.1) and spayed females (OR 2.3) were predominant compared to intact dogs and intact females, respectively. Ulceration (OR 5.2) and lymph node metastasis (OR 7.1) occurred more frequently in larger MCTs. Both ulceration and MCTs > 3 cm were highly associated with lymph node metastasis (OR 24.8). Recurrence was associated with MCT-related death (OR 10.50, p = 0.0040), and the latter was associated with shorter survival times (p = 0.0115). Dogs with MCTs > 3 cm (p = 0.0040), lymph node metastasis (p = 0.0234), or elevated WHO stage (p = 0.0158) had shorter survival times. A significantly higher frequency of MCTs was found in specific breeds, and in older and neutered dogs. MCTs > 3 cm and lymph node or distant metastases were associated with shorter survival times.

ACVIM consensus statement on the diagnosis of immune-mediated hemolytic anemia in dogs and cats.

Immune-mediated hemolytic anemia (IMHA) is an important cause of morbidity and mortality in dogs. IMHA also occurs in cats, although less commonly. IMHA is considered secondary when it can be attributed to an underlying disease, and as primary (idiopathic) if no cause is found. Eliminating diseases that cause IMHA may attenuate or stop immune-mediated erythrocyte destruction, and adverse consequences of long-term immunosuppressive treatment can be avoided. Infections, cancer, drugs, vaccines, and inflammatory processes may be underlying causes of IMHA. Evidence for these comorbidities has not been systematically evaluated, rendering evidence-based decisions difficult. We identified and extracted data from studies published in the veterinary literature and developed a novel tool for evaluation of evidence quality, using it to assess study design, diagnostic criteria for IMHA, comorbidities, and causality. Succinct evidence summary statements were written, along with screening recommendations. Statements were refined by conducting 3 iterations of Delphi review with panel and task force members. Commentary was solicited from several professional bodies to maximize clinical applicability before the recommendations were submitted. The resulting document is intended to provide clinical guidelines for diagnosis of, and underlying disease screening for, IMHA in dogs and cats. These should be implemented with consideration of animal, owner, and geographical factors.

Soft tissue sarcoma in dogs: A treatment review and a novel approach using electrochemotherapy in a case series.

Canine soft tissue sarcomas (STSs) are locally invasive mesenchymal neoplasms. Electrochemotherapy (ECT) is an antitumour local ablative treatment that uses electric pulses to enhance the intracellular delivery of cytotoxic drugs. The aim of this retrospective study was to review the current treatment for STSs and to evaluate the efficacy and safety of ECT with bleomycin in canine STSs. Fifty-two dogs with 54 STSs were included. Three groups were arranged: (a) ECT alone, (b) intra-operative ECT and (c) adjuvant ECT. Signalment, tumour size, location, histological grade and margins and ECT parameters were collected. Recurrence rate (RR) and disease-free interval (DFI) were calculated. Treatment toxicity was assessed using a 6-point scale. STSs were mostly located on limbs (77.8%). Median tumour size was 4.3 cm (range 0.4-17.0 cm). Most STSs were grade I (47.7%) and II (50.0%), and histological margins were incomplete in 94.5% of cases. Two complete remissions, one partial remission and one stable disease were recorded in group 1. Group 2 and 3 were similar for tumour location, size and grade, histological margins, treatment toxicity, pulse frequency and voltage. Moreover, RR and DFI were similar between group 2 and 3 (23% and 25%, 81.5 and 243 days, respectively). Local toxicity post ECT was mild (score ≤ 2) in 66.7% of cases. Higher toxicity score was associated with higher pulse voltage (1200 vs 1000 V/cm) (P = 0.0473). ECT coupled with bleomycin resulted safe and efficient in tumour local control and should be considered as an option for treatment of canine STSs.

Tel-eVax: a genetic vaccine targeting telomerase for treatment of canine lymphoma.

BACKGROUND: we have recently shown that Tel-eVax, a genetic vaccine targeting dog telomerase (dTERT) and based on Adenovirus (Ad)/DNA Electro-Gene-Transfer (DNA-EGT) technology can induce strong immune response and increase overall survival (OS) of dogs affected by multicentric Diffuse Large B cell Lymphoma (DLBCL) when combined to COP therapy in a double-arm study. Here, we have utilized a clinically validated device for veterinary electroporation called Vet-ePorator™, based on Cliniporator™ technology currently utilized and approved in Europe for electrochemotherapy applications and adapted to electrogenetransfer (EGT). METHODS: 17 dogs affected by DLBCL were vaccinated using two Ad vector injections (Prime phase) followed by DNA-EGT (Boost phase) by means of a Vet-ePorator™ device and treated in the same time with a 27-week Madison Wisconsin CHOP protocol. The immune response was measured by ELISA assays using pool of peptides. RESULTS: No significant adverse effects were observed. The OS of vaccine/CHOP animals was 64.5 weeks, in line with the previous study. Dogs developed antibodies against the immunizing antigen. CONCLUSIONS: Tel-eVax in combination with CHOP is safe and immunogenic in lymphoma canine patients. These data confirm the therapeutic efficacy of dTERT vaccine and hold promise for the treatment of dogs affected by other cancer types. More importantly, our findings may translate to human clinical trials and represent new strategies for cancer treatment.

Neospora caninum oocyst shedding in a naturally infected dog from Italy.

Although the seroprevalence of Neospora caninum infection in dogs can be relatively high, there are few reports of dogs naturally shedding N. caninum oocysts. Worldwide, the prevalence of Neospora excretion in canine faeces ranges from 0.03% to 4.9%. A mixed-breed male household dog of about 8years in age living in the district of Pisa (Tuscany, Central Italy) was referred for dysorexia, weakness and general lymph node enlargement. Clinical pathology demonstrated mild normocytic and normochromic anemia, thrombocytopenia and hypoproteinemia with hypoalbuminemia. Serology for Leishmania, Ehrlichia canis and Anaplasma phagocytophilum was negative. From lymph node and bone marrow analysis, T cell lymphoma, high grade, pleomorphic type, clinical stage V, was diagnosed. The dog was treated with a chemotherapy induction protocol with vincristine, cyclophosphamide, and prednisone for 8weeks. A faecal sample collected from the dog 7days after the beginning of the treatment and analysed by flotation test and a McMaster method, revealed the presence of 300 OPG N. caninum-like unsporulated oocysts of about 10-11µm in diameter. An aliquot of the same faecal sample analysed by PCR with species-specific primer pairs Np6+/Np21+ was positive for N. caninum DNA, while specific serology performed on sera collected at the first visit and a month later by IFAT, were positive with a titer of 1: 50 and 1:400, respectively. Soon after the dog died. Naturally occurring systemic illness or iatrogenic immunosuppression may predispose dogs to proliferation of the parasite. The dog was receiving chemo-immunosuppressive treatment for T cell lymphoma. For this reason it is possible to suppose that emission of Neospora oocysts in this dog was caused by reactivation of a latent infection.

Safety and efficacy of a genetic vaccine targeting telomerase plus chemotherapy for the therapy of canine B-cell lymphoma.

Client-owned pet dogs represent exceptional translational models for advancement of cancer research because they reflect the complex heterogeneity observed in human cancer. We have recently shown that a genetic vaccine targeting dog telomerase reverse transcriptase (dTERT) and based on adenovirus DNA electro-gene-transfer (Ad/DNA-EGT) technology can induce strong cell-mediated immune responses against this tumor antigen and increase overall survival of dogs affected by B-cell lymphosarcoma (LSA) in comparison with historical controls when combined with a cyclophosphamide, vincristine, and prednisone (COP) chemotherapy regimen. Here, we have conducted a double-arm clinical trial with an extended number of LSA patients, measured the antigen-specific immune response, and evaluated potential toxic effects of the immunotherapy along with a follow-up of patients survival for 3.5 years. The immune response was measured by enzyme-linked immunospot assay. The expression of dTERT was quantified by quantitative polymerase chain reaction. Changes in hematological parameters, local/systemic toxicity or organic dysfunction and fever were monitored over time during the treatment. dTERT-specific cell-mediated immune responses were induced in almost all treated animals. No adverse effects were observed in any dog patient that underwent treatment. The overall survival time of vaccine/COP-treated dogs was significantly increased over the COP-only cohort (>76.1 vs. 29.3 weeks, respectively, p<0.0001). There was a significant association between dTERT expression levels in LSA cells and overall survival among vaccinated patients. In conclusion, Ad/DNA-EGT-based cancer vaccine against dTERT in combination with COP chemotherapy is safe and significantly prolongs the survival of LSA canine patients. These data confirm the therapeutic efficacy of dTERT vaccine and support the evaluation of this approach for other cancer types as well as the translation of this approach to human clinical trials.

Use of totally implantable vascular access port with mini-invasive Seldinger technique in 12 dogs undergoing chemotherapy.

Vascular access ports (VPAs) are totally implantable devices designed to provide repeated access to the vascular system. Port access is performed by percutaneous needle insertion using a noncoring needle. VAPs have been placed in 12 dogs affected by different tumors and needing long lasting chemotherapy. Using the non-invasive Seldinger technique a silicone catheter was inserted from the jugular vein up to the junction of the cranial vena cava and the right atrium. The catheter then was connected to the port previously placed in a subcutaneous pocket over the scapula. 7/12 dogs showed no clinical complications. Port was removed in 4/12 dogs for post-operative complications as fistula formation (n.2) and infection/mal-position (n.2). One patient had mild complications shortly after implantation. VPAs were left in site until the death in the rest of patients. VAPs are useful for dogs undergoing long-lasting chemotherapy protocols as these devices allow peripheral veins to be spared and not seriously damaged for repeated infusion of vesicant drugs. In most cases VPAs are well tolerated and may be left in site for several months.

Monitoring the reverse to normal of clinico-pathological findings and the disease free interval time using four different treatment protocols for canine leishmaniosis in an endemic area.

Twenty-four dogs naturally infected by Leishmania spp. were treated with four different protocols using meglumine antimoniate (aNm) and allopurinol in combination or in monotherapy. Aiming to compare the efficacy of the different treatments the reverse to normal of clinico-pathological findings and the disease free interval time (DFIT) were evaluated. Treated dogs were monitored for 1 year and, in absence of relapses, the DFIT was postponed to the last available follow-up. Seven dogs treated with aNm alone showed relapses during the year of observation. In the group of dogs treated with the combination of aNm (50 mg/kg/SC 12 hourly up to clinico-pathological recovery) and allopurinol (15 mg/kg/PO 12 hourly administered for 6months) no relapses were registered in the year of monitoring and the DFIT reached up to 65 months. Our results showed that this combination represents the best choice to treat canine leishmaniosis compared to other protocols.

Evaluation of erythrocytes, platelets, and serum iron profile in dogs with chronic enteropathy.

The aim of this study is to evaluate iron status, erythrocyte, and platelet modifications in dogs with chronic enteropathy (CE). Dogs were grouped as food-responsive diarrhea (FRD, n = 11), antibiotic-responsive diarrhea (ARD, n = 5), and steroid-responsive diarrhea (SRD, n = 6) relating to therapeutic-response. Clinical and haematological findings, evidence of gastrointestinal blood loss, and iron metabolism were evaluated before and after treatment. A mild normocytic or microcytic anemia and thrombocytosis were identified, respectively in 18.0% and 31.8% of CE dogs. No significant differences between pre- and posttreatment of hematocrit, haemoglobin, and mean corpuscular volume, platelet count and mean platelet volume were found. Statistical analysis pointed out significant differences between pre- and posttreatment in serum iron (P < .03) and unsaturated iron binding capacity (UIBC) (P < .01). No significant correlations were found between these parameters and canine Inflammatory Bowel Disease activity index and pattern of CE as well.

A vaccine targeting telomerase enhances survival of dogs affected by B-cell lymphoma.

Canine cancers occur with an incidence similar to that of humans and share many features with human malignancies including histological appearance, tumor genetics, biological behavior, and response to conventional therapies. As observed in humans, the telomerase reverse transcriptase (TERT) activity is largely confined to tumor tissues and absent in the majority of normal dog tissues. Therefore, dog TERT (dTERT) can constitute a valid target for translational cancer immunotherapy. We have evaluated the ability of adenovirus serotype 6 (Ad6) and DNA electroporation (DNA-EP) to induce immune responses against dTERT in dogs affected by malignant lymphoma (ML). The vaccine was combined with standard chemotherapy regimen [cyclophosphamide, vincristine, prednisone (COP)]. dTERT-specific immune response was induced in 13 out of 14 treated animals (93%) and remained detectable and long-lasting with the absence of autoimmunity or other side effects. Most interestingly, the survival time of vaccine/Chemo-treated dogs was significantly increased over historic controls of Chemo-treated animals (>97.8 versus 37 weeks, respectively, P = 0.001). Our results show that Ad6/DNA-EP-based cancer vaccine against dTERT overcomes host immune tolerance, should be combined with chemotherapy, induces long-lasting immune responses, and significantly prolongs the survival of ML canine patients. These data support further evaluation of this approach in human clinical trials.

A retrospective study of 60 cases of eccentrocytosis in the dog.

BACKGROUND: Eccentrocytes are RBCs that appear in a peripheral blood smear to have their hemoglobin shifted to one side of the cell. This abnormality, which is confined to the RBC membrane and cytoskeleton, is induced by oxidative damage. Eccentrocytes have been reported rarely in dogs and are associated with onion and garlic ingestion and the administration of oxidant drugs. OBJECTIVE: The purpose of this study was to describe the occurrence and severity of eccentrocytosis in dogs and the diseases or disorders associated with eccentrocytes. METHODS: Detailed history, and results of physical examination, CBC, biochemical and coagulation profiles, and urinalysis from all canine patients admitted during a 2.5-year period were evaluated. Eccentrocytes, when observed, were graded 1+ (few) to 4+ (many). The severity of eccentrocytosis was compared with that of anemia and reticulocytosis. RESULTS: Eccentrocytes were found in blood smears from 60 of 4251 dogs (1.4%) and were associated with mild to moderate anemia in 40 (66.6%) of the cases. Eccentrocytosis was found in 16 (26.6%) dogs with drug administration, 11 (18.3%) with presumptive onion and garlic ingestion, 8 (13.3%) with vitamin K antagonist intoxication, 7 (11.6%) with ketoacidotic diabetes, 5 (8.3%) with T-cell lymphoma, 4 (6.6%) with severe infections, 1 (1.6%) with compensated diabetes mellitus, and 8 (13.3%) with other conditions. Certain dog breeds, such as Whippet, Boxer, and English Setter, and young dogs seemed to be overrepresented. CONCLUSIONS: We describe, for the first time, associations between eccentrocytes and diabetes mellitus, T-cell lymphoma and vitamin K antagonist intoxication in dogs. The significance of eccentrocytes should not be underestimated, because they can be a signal of an oxidative process.