BackgroundBreast (mammary) cancers in human (BC) and canine (CMT) patients share clinical, pathological, and molecular similarities that suggest dogs may be a useful translational model. Many cancers, including BC, shed exosomes that contain microRNAs (miRs) into the microenvironment and circulation, and these may represent biomarkers of metastasis and tumor phenotype.MethodsThree normal canine mammary epithelial cell (CMEC) cultures and 5 CMT cell lines were grown in serum-free media. Exosomes were isolated from culture media by ultracentrifugation then profiled by transmission electron microscopy, dynamic light scattering, and Western blot. Exosomal small RNA was deep-sequenced on an Illumina HiSeq2500 sequencer and validated by qRT-PCR. In silico bioinformatic analysis was carried out to determine microRNA gene and pathway targets.ResultsCMEC and CMT cell lines shed round, “cup-shaped” exosomes approximately 150–200 nm, and were immunopositive for exosomal marker CD9. Deep-sequencing averaged ~ 15 million reads/sample. Three hundred thirty-eight unique miRs were detected, with 145 having > ±1.5-fold difference between one or more CMT and CMEC samples. Gene ontology analysis revealed that the upregulated miRs in this exosomal population regulate a number of relevant oncogenic networks. Several miRNAs including miR-18a, miR-19a and miR-181a were predicted in silico to target the canine estrogen receptor (ESR1α).ConclusionsCMEC and CMT cells shed exosomes in vitro that contain differentially expressed miRs. CMT exosomal RNA expresses a limited number of miRs that are up-regulated relative to CMEC, and these are predicted to target biologically relevant hormone receptors and oncogenic pathways. These results may inform future studies of circulating exosomes and the utility of miRs as biomarkers of breast cancer in women and dogs.Electronic supplementary materialThe online version of this article (10.1186/s12885-018-4750-6) contains supplementary material, which is available to authorized users.

Objectives The objectives of this study were to determine the prevalence of feline coronavirus (FCoV) viremia, and its replication in peripheral blood using quantitative RT-PCR (qRT-PCR) methodology in a population of 205 healthy shelter cats in Southern California, as well as to assess any possible connection to longitudinal development of feline infectious peritonitis (FIP). Methods The study was performed on buffy-coat samples from EDTA-anticoagulated whole blood samples of 205 healthy shelter cats. From 50 of these cats, fecal samples were also examined. FCoV genomic and subgenomic RNA in the buffy coats was amplified by a total FCoV RNA qRT-PCR. Evidence for FCoV replication in peripheral blood and feces was obtained by M gene mRNA qRT-PCR. Results Nine of 205 cats (4.4%) were viremic by the total FCoV RNA qRT-PCR, and one of these cats had evidence of peripheral FCoV blood replication by an FCoV mRNA qRT-PCR. The single cat with peripheral blood replication had a unique partial M gene sequence distinct from positive controls and previously published FCoV sequences. Neither seven of the nine viremic cats with follow-up nor the single cat with replicating FCoV with positive qRT-PCR results developed signs compatible with FIP within 6 months of sample collection. Conclusions and relevance FCoV viremia and peripheral blood replication in healthy shelter cats have a low prevalence and do not correlate with later development of FIP in this study population, but larger case-control studies evaluating the prognostic accuracy of the qRT-PCR assays are needed.

Objective:To compare the prognostic value of admission hematologic parameters serum/plasma iron, red blood cell distribution width (RDW), and nucleated red blood cells (nRBCs) in dogs presenting with acute traumatic injury.Design: Retrospective observational study (2009)(2010)(2011)(2012)(2013)(2014)(2015).Setting: University teaching hospital. Animals:One hundred and twenty-nine clinical dogs presenting within 24 hours of acute traumatic injury. Interventions: None.Measurements and Main Results: One hundred and twenty-nine dogs met the inclusion criteria and 109 (84.5%) survived, while 20 (15.5%) died or were euthanized in hospital. Patients with blunt force trauma comprised 79.8% of the patient population; dogs with penetrating trauma comprised 20.2% of cases. Hypoferremia occurred in all nonsurvivors, and the median serum/plasma iron concentration was significantly lower in nonsurvivors than survivors (P = 0.028). Normal or increased serum/plasma iron had 100% specificity and 100% positive predictive value for survival.Red blood cell distribution width was not significantly different between groups (P = 0.417). The presence of nRBCs was significantly associated with nonsurvival (P = 0.030), although the absolute nRBC concentrations were not significantly different (P = 0.070). A multiple logistic regression model found age, type of injury, presence of nRBCs, and serum/plasma iron to be independent predictors of survival with an area under the receiver operator characteristic curve of 0.813. Conclusions:The presence of nRBCs and low serum/plasma iron are associated with mortality in patients with acute trauma; however, red blood cell distribution width was not associated with survival. Absence of hypoferremia was highly associated with a favorable prognosis in this patient population. These parameters may warrant inclusion in trauma scoring systems. K E Y W O R D SnRBCs, nucleated red blood cells, RDW, serum/plasma iron, trauma Abbreviations: APPLE FAST , acute patient physiologic and laboratory evaluation score ( FAST = 5 parameter score, FULL = 10 parameter score); ATT, Animal Trauma Triage; nRBCs, nucleated red blood cells; RDW, red blood cell distribution width; ROC, receiver operating characteristic; SIRS, systemic inflammatory response syndrome.

Background: Differentiating benign from canine malignant mammary tumors requires invasive surgical biopsy. Circulating microRNAs (miRNA) may represent promising minimally invasive cancer biomarkers in people and animals.Objectives: To evaluate the serum mRNA profile between dogs with and without mammary carcinoma, and to determine if any of these markers have prognostic significance.Animals: Ten healthy client-owned female dogs (5 intact, 5 spayed) and 10 dogs with histologically confirmed mammary carcinoma were included; 9 were client-owned, whereas 1 was a research colony dog.Methods: Retrospective study. Serum miRNA was evaluated by RNA deep-sequencing (RNAseq) and digital droplet PCR (dPCR).Expression of candidate biomarkers miR-18a, miR-19b, miR-29b, miR-34c, miR-122, miR-125a, and miR-181a was compared with clinical characteristics, including grade, metastasis, and survival.Results: 452 unique serum miRNAs were detected by RNAseq. Sixty-five individual miRNAs were differentially expressed (>±1.5-fold) and statistically significant between groups. Serum miR-19b (P = .003) and miR-125a (P < .001) were significantly higher in the mammary carcinoma group by dPCR. Both had high accuracy based on receiver operator characteristic area under the curve (0.930 for miR-125a; 0.880 for miR-19b). Circulating miR-18a by RNAseq was significantly higher in mammary carcinoma dogs with histologic evidence of lymphatic invasion (P = 0.03). There was no significant association with any miRNA and survival or inflammatory status. Conclusions and Clinical Importance: Circulating miRNAs are differentially expressed in dogs with mammary carcinoma. Serum miR-19b and miR-18a represent candidate biomarkers for diagnosis and prognosis, respectively. Abbreviations: AUC, area under the curve; CMT, canine mammary tumor(s); dPCR, digital droplet PCR; FAM, 6-carboxyfluorescein; miRs/miRNA, microRNA; qPCR, quantitative PCR; RNAseq, RNA deep-sequencing; ROC, receiver operator characteristic.

An 8-year-old, female spayed Domestic Shorthair cat was presented to the Auburn University Emergency and Critical Care service for evaluation of pleural effusion and a suspected intrathoracic mass. Computed tomography was performed which confirmed the presence of a large intrathoracic mass, likely heart-based. Fine-needle aspirates were obtained and a cytologic diagnosis of a neuroendocrine tumor was made. Treatment with toceranib phosphate was briefly attempted at home by the owners. The cat died at home approximately 6 weeks after diagnosis. Necropsy and subsequent histopathologic examination revealed a metastatic neuroendocrine carcinoma of aortic body origin. Aortic body tumors are extremely rare in cats and to the authors' knowledge, a neuroendocrine carcinoma of aortic body origin with distant metastases has not yet been reported in a cat.

Introduction: Hyperbaric oxygen therapy (HBOT) involves breathing 100% oxygen in a specialized compression chamber leading to hyperoxia. This treatment modality is associated with anti-inflammatory, antioxidant, and healing properties in people and laboratory animals. However, there are relatively few reports that evaluate the effects of HBOT in companion animals. The goal of this study was to investigate the physiological effects of HBOT on surgically induced systemic inflammation and oxidation in dogs.Material and Methods: Twelve healthy female beagle dogs were spayed and randomized into control and HBOT groups (n = 6). Both groups received conventional post-ovariohysterectomy therapy, and the HBOT group received two hyperbaric treatments at 2.0 atmosphere of absolute pressure and 100% oxygen for 35 min, 6 and 18 h after surgery. Blood samples were collected 3 h prior to ovariohysterectomy, 6, 18, and 30 h after surgery, prior to HBOT when applicable. Inflammatory biomarkers, including C-reactive protein, circulating cytokines, and changes in iron homeostasis were evaluated at each time point to determine the effects of surgery and HBOT on inflammation. Similarly, serum total oxidant status and total antioxidant status were measured to assess the oxidative stress. Pain and incision scores were recorded and compared between groups.Results: Following ovariohysterectomy, all dogs had significantly increased serum concentrations of C-reactive protein, KC-like, IL-6, and increased unsaturated ironbinding capacity compared to their pre-surgical values (p < 0.02), while serum iron, total iron-binding capacity and transferrin saturation were significantly decreased after surgery (p < 0.02). There was no significant difference between the control group and the HBOT group for any of the variables. There were no overt adverse effects in the HBOT group. Conclusion:This is the first prospective randomized controlled study to investigate the effects of HBOT on surgically induced systemic inflammation in dogs. While elective ovariohysterectomy resulted in mild inflammation, the described HBOT protocol Gautier et al.Ovariohysterectomy and HBOT in Dogs portrayed no outward adverse effect and did not induce any detectable pro-inflammatory, anti-inflammatory, or antioxidant effects. Additional investigation is required to identify objective markers to quantify the response to HBOT and determine its role as an adjunctive therapy in dogs with more severe, complicated or chronic diseases.

An 8-year-old female spayed Weimaraner dog was presented to the Bailey Small Animal Teaching Hospital at Auburn University for evaluation of ascites and lethargy. On physical examination, the dog was approximately 7% dehydrated, had a distended abdomen, a palpable fluid wave, and mild cranial organomegaly. Serum biochemistry results revealed mild hyperbilirubinemia (0.32 mg/dL, reference interval [RI] 0-0.2 mg/dL), hypocholesterolemia (121 mg/dL, RI 132-335 mg/dL), hyponatremia (138 mmol/L, RI 142-151 mmol/L), and hypochloremia (96 mmol/L, RI 105-117 mmol/L); all other results were within the RI. Complete blood count results were unremarkable. Thoracic radiographs did not reveal any abnormalities.An abdominal ultrasound examination revealed abundant abdominal fluid and multiple hypoechoic to targetlike splenic masses. The spleen was considered normal in size and shape. Fine-needle aspirate (FNA) cytology of the splenic masses was poorly cellular. The effusion was pink and opaque, with a nucleated cell concentration of 15,340/ lL, an erythrocyte concentration of 340,000/lL, and a refractometric total protein concentration of 8.4 g/dL (protein concentration likely spuriously increased due to turbidity). Direct smears of the abdominal effusion were examined (Figure 1). Abdominal contrast computed tomography (CT) scan and lymphangiography did not reveal a cause for the effusion, so an exploratory laparotomy was performed. At surgery, the liver was small, and dark red-black nodules were noted in the spleen, liver, omentum, and outer aorta. Impression smears of nodules were examined (Figure 2). A B Figure 2. Impression smears of a splenic nodule of a dog. Modified Wrights. (A) 920 objective; (B) 950 objective.Figure 1. Direct smear of abdominal fluid from a dog. Modified Wrights. 9100 objective.

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