Prolonged holding time and sampling protocol affects viscoelastic coagulation parameters as measured by the VCM-Vet™ using fresh equine native whole blood.

OBJECTIVES: Determine the effect of sample holding time and single sample reuse on viscoelastic coagulation parameters when using fresh equine native whole blood. ANIMALS: 8 healthy adult horses from a university teaching herd. PROCEDURES: Blood collected by direct jugular venipuncture (18 ga needle, 3 mL syringe) was held at 37 °C for 2, 4, 6, or 8 minutes according to 1 of 2 protocols. Syringes were gently inverted twice, a small amount of blood was expressed, testing cartridges were filled, and placed within the VCM-Vet™ device (Entegrion Inc). Protocol A: samples were processed from a single syringe. Protocol B: 4 syringes were drawn through a single needle. VCM-Vet™ measures assessed included clot time (CT), clot formation time (CFT), alpha angle (AA), amplitude at 10/20 minutes (A10/A20), maximal clot firmness (MCF), and lysis index at 30/45 minutes (LI30/LI45). Differences over time were examined using the Friedman test and post hoc Wilcoxon Rank Sum Test with Bonferroni correction, P ≤ .05. RESULTS: Following Protocol A, there was a significant effect of holding time for CT (P = .02), CFT (P = .04), and AA (P = .05). CT and AA decreased over time, while CFT increased. Samples handled by Protocol B showed no significant difference over time for any of the VCM-Vet™ parameters. CLINICAL RELEVANCE: Sample holding time and handling protocol impact VCM-Vet™ testing results of fresh equine native whole blood. Viscoelastic coagulation samples tested using the VCM-Vet™ may be held unagitated for up to 8 minutes after collection while warm, but should not be reused.

Acute phase protein concentrations following serial procaine penicillin G injections in horses.

BACKGROUND: Acute phase protein (APP) measurement is used to detect inflammation. Intramuscular (IM) injections could cause tissue injury and induce an acute phase response (APR). OBJECTIVES: To evaluate the effects of IM procaine penicillin G (PPG) injections on APP concentrations in horses. STUDY DESIGN: Prospective longitudinal design. METHODS: PPG was administered intramuscularly to six horses, twice daily, for 5 days. Plasma fibrinogen (FIB), serum amyloid A (SAA), haptoglobin (HAP), creatine kinase (CK), and aspartate aminotransferase (AST) were quantified daily for 5 days before the first injection, during the course of administration, and for 4 days after the final dose. Analytes were quantified every other day for the remaining 16 days. Data were compared using a parametric or non-parametric repeated measures ANOVA and a Tukey's or Mann-Whitney rank sum test, respectively. Significance was set at p < 0.05. RESULTS: CK was increased over baseline (mean ± SD: 200 ± 74 IU/L) on Days 1-6 (p < 0.001 to p = 0.02, mean ± SD: 723-1177 ± 355-544 IU/L) and AST was increased above baseline (mean ± SD: 233 ± 58 IU/L) on Days 2-7 and 10 (p < 0.001 to p = 0.05, mean ± SD: 307-437 ± 79-146 IU/L). Increased FIB was noted over baseline (mean ± SD: 177 ± 30 mg/dl) on Days 6-8 and 10 (p = 0.02 to p = 0.03, mean ± SD: 234-252 ± 33-49 mg/dl). SAA was increased above baseline (mean ± SD: 4.7 ± 2.9) on Day 6 (p = 0.02, mean ± SD: 113 ± 186 µg/ml). There was no change in HAP. MAIN LIMITATIONS: Healthy horses were used, small sample size, and a lack of a negative control group. CONCLUSIONS: Serial intramuscular procaine penicillin G (IM PPG) injections may result in increased positive APP concentrations in horses and this must be considered when these test results are interpreted.

Vaginal swab cytology as a diagnostic tool for neoplasia of the lower urinary tract in 5 dogs.

Urothelial carcinoma (UC), or transitional cell carcinoma, is the most common canine urothelial malignancy in dogs. Females are predisposed and provide a challenge for diagnostic tumor sampling. The objectives of this study were to investigate the use and tolerability of vaginal swab cytology for UC diagnosis. Five dogs were identified with non-diagnostic urine sediment cytology and UC diagnosed on vaginal cytology with confirmation by another means. All patients tolerated the vaginal swab with minimal restraint. This study confirms the potential of vaginal swab cytology as a simple, inexpensive, and well-tolerated means for lower urinary tract UC diagnosis in female dogs. Key clinical message: Vaginal swab cytology is a non-invasive, low-cost method of obtaining a sample for cytological assessment for UC.

La cytologie vaginal par écouvillonnage comme outil de diagnostic de la néoplasie des voies urinaires inférieures chez cinq chiennes. Le carcinome urothélial (CU), ou carcinome à cellules transitionnelles, est la tumeur maligne urothéliale canine la plus courante. Les femelles sont prédisposées et constituent un défi pour l'échantillonnage diagnostique des tumeurs. Les objectifs de cette étude étaient d'étudier l'utilisation et la tolérabilité de la cytologie vaginale par écouvillonnage pour le diagnostic du CU. Cinq chiennes ont été identifiées avec une cytologie des sédiments urinaires non diagnostique et un CU diagnostiqué sur la cytologie vaginale avec confirmation par un autre moyen. Toutes les patientes ont toléré le prélèvement vaginal avec un minimum de contention. Cette étude confirme le potentiel de la cytologie vaginale par écouvillonnage en tant que moyen simple, peu coûteux et bien toléré pour le diagnostic du CU des voies urinaires inférieures chez les chiennes.Message clinique clé :La cytologie vaginale par écouvillonnage est une méthode non-invasive et peu coûteuse pour obtenir un échantillon pour évaluation cytologique du CU.(Traduit par les auteurs).

Hyponatremia in horses with septic pneumopathy.

BACKGROUND: Hyponatremia is common in horses with bacterial pleuropneumonia, but no further characterization of this abnormality has been reported. OBJECTIVES: Describe admission plasma sodium concentration ([Na]) in horses with septic pneumopathy and evaluate any association of plasma [Na] with markers of systemic inflammation. ANIMALS: Medical records of horses >1 month of age that between 2008 and 2021 had a transtracheal aspirate (TTA) performed, abnormal TTA cytology, positive TTA culture, pulmonary disease on ultrasonography, radiography or both, and plasma [Na] assessed by direct ion-selective-electrode (dISE). Horses with concurrent diarrhea or azotemia were excluded. METHODS: Clinical and clinicopathological variables of interest between hypo- and normonatremic horses were compared. Spearman correlation and Fisher exact tests were used to identify significant associations (P < .05). RESULTS: Twenty of 35 horses had hyponatremia (median, 132 mmol/L; 25-75th interquartile range [IQR], 129.7-133.1 mmol/L; reference range, 134.2-138.4 mmol/L). A higher proportion of horses with systemic inflammatory response syndrome (SIRS) had hyponatremia (P = .01). Hyponatremic patients had higher mean plasma fibrinogen concentration (461 ± 160.5 mg/dL; P = .01) and higher rectal temperature (38.8 ± 0.7°C; P = .02) than normonatremic horses. Negative correlations were found between plasma [Na] and fibrinogen (P = .001; ρ = -0.57) concentrations and between plasma [Na] and rectal temperature (P = .001; ρ = -0.51). Presence or absence of pleural effusion did not influence severity of hyponatremia. Mean duration of hospitalization was longer (P = .04) in hyponatremic horses (9.8 ± 6.6 days). CONCLUSIONS AND CLINICAL IMPORTANCE: Hyponatremia at admission is associated with the presence of inflammation, SIRS, and with longer duration of hospitalization.

Total protein concentration as a predictor of neoplastic peritoneal and pleural effusions of dogs.

BACKGROUND: The diagnosis of neoplastic cavitary effusions requires the identification of neoplastic cells in effusions, yet the cytologic appearance of neoplastic effusions can be highly variable due to the varied mechanisms of formation. Additional parameters might aid in the interpretation of equivocal cytologic results. OBJECTIVES: Our goal was to evaluate whether total protein concentrations can be used to support the diagnosis of neoplasia in the peritoneal and pleural effusions of dogs with lower cellularities (≤5000 nucleated cells/µL). METHODS: Pleural and peritoneal fluid analyses from dogs presented to the University of Illinois Veterinary Teaching Hospital between 2014 and 2019 were evaluated retrospectively. Effusions were categorized as neoplastic or non-neoplastic based on histology or cytology. Non-neoplastic effusions were subcategorized according to mechanism: decreased oncotic pressure, increased hydrostatic pressure, increased vascular permeability, leakage of urine, and leakage of lymph. The TP and blood albumin to fluid TP ratio (Albblood :TPfluid ) were compared among groups. RESULTS: Twenty-seven neoplastic and 65 non-neoplastic cases were evaluated. TP was higher in the neoplastic group (P = .001) than in the non-neoplastic group. Neoplastic effusions had a lower Albblood :TPfluid than non-neoplastic (P = .001), and effusions with Albblood :TPfluid of ≤0.6 were 5.6 times more likely to be neoplastic (95% CI 1.69-17.36; P = .003). CONCLUSIONS: Fluid TP concentrations were significantly greater in neoplastic than non-neoplastic effusions; however, given the considerable overlap between groups, the diagnostic utility of this difference is low. A neoplastic etiology might be more likely in cases with an Albblood :TPfluid ≤0.6.

The use of alkaline phosphatase and runx2 to distinguish osteosarcoma from other common malignant primary bone tumors in dogs.

In dogs, primary bone tumors can be difficult to distinguish with histopathology. Of those tumors, osteosarcoma (OSA) is the most common and aggressive. In this study, 4 immunohistochemistry markers-alkaline phosphatase (ALP), osteonectin (ON), osteopontin (OP), and runx2-were evaluated for their ability to distinguish OSA from other primary bone tumors. The 42 formalin-fixed, paraffin-embedded, primary canine bone tumors included 15 OSAs, 8 chondrosarcomas, 11 fibrosarcomas, and 8 histiocytic sarcomas. All 4 antibodies were highly sensitive for detection of osteosarcoma. ALP was the most sensitive at 100% and runx2 the most specific at 78%. Running ALP and runx2 in series resulted in a sensitivity of 87% and a specificity of 85%. This combination of immunomarkers resulted in a diagnostic panel for distinguishing osteosarcoma from other primary bone tumors.

Blood-to-saliva glucose time lag in sedated healthy dogs.

The management of diabetes mellitus mandates measurement of blood glucose. Saliva offers an alternative to blood sampling, but measurement of the salivary glucose concentration is difficult, and the blood-to-saliva glucose time lag is uncertain. We aimed to determine the serum-saliva glucose time lag in the saliva of healthy dogs. The combined duct of the mandibular and sublingual salivary glands of 6 dogs was cannulated to collect saliva and prevent glucose degradation by oral bacteria. Following a 0.25 g/kg IV bolus of dextrose, paired serum-saliva samples were collected at baseline and in twelve 5-min blocks over 60 min. Serum and salivary glucose levels were analyzed with a linear mixed model for repeated measures with a compound symmetry error structure. Mean (±SD) saliva production was 10.3 ± 2.9 µL/kg/min, and the area under the curve (AUCglucose)saliva/serum ratio was 0.006, which highlights the magnitude of the large difference in glucose concentration between the 2 compartments. The serum-saliva glucose time lag was 30-40 min.

Utilizing feline oral squamous cell carcinoma patients to develop NQO1-targeted therapy.

Developing effective therapies for the treatment of advanced head-and-neck squamous cell carcinoma (HNSCC) remains a major challenge, and there is a limited landscape of effective targeted therapies on the horizon. NAD(P)H:quinone oxidoreductase 1 (NQO1) is a 2-electron reductase that is overexpressed in HNSCC and presents as a promising target for the treatment of HNSCC. Current NQO1-targeted drugs are hindered by their poor oxidative tolerability in human patients, underscoring a need for better preclinical screening for oxidative toxicities for NQO1-bioactivated small molecules. Herein, we describe our work to include felines and feline oral squamous cell carcinoma (FOSCC) patients in the preclinical assessment process to prioritize lead compounds with increased tolerability and efficacy prior to full human translation. Specifically, our data demonstrate that IB-DNQ, an NQO1-targeted small molecule, is well-tolerated in FOSCC patients and shows promising initial efficacy against FOSCC tumors in proof-of-concept single agent and radiotherapy combination cohorts. Furthermore, FOSCC tumors are amenable to evaluating a variety of target-inducible couplet hypotheses, evidenced herein with modulation of NQO1 levels with palliative radiotherapy. The use of felines and their naturally-occurring tumors provide an intriguing, often underutilized tool for preclinical drug development for NQO1-targeted approaches and has broader applications for the evaluation of other anticancer strategies.

Comparison of creatine kinase in cerebrospinal fluid collected from the cerebellomedullary and lumbar cisterna in 10 dogs with neurologic disease.

BACKGROUND: Analysis of cerebrospinal fluid CK (CSF-CK) might be useful as a prognostic indicator in dogs with neurologic disease. Previous studies have mostly analyzed CSF-CK collected from the cerebellomedullary (CM) cisterna, but CSF collection sites could affect its levels. OBJECTIVES: This is a pilot study aimed to evaluate differences in CSF-CK concentrations when collected from the CM or lumbar cisterna in dogs presenting with neurologic disease. METHODS: Ten dogs presenting for neurologic disease underwent magnetic resonance imaging and CSF collection from both the CM and lumbar cisterna. Cerebrospinal fluid CK was analyzed within 30 minutes. RESULTS: Ten dogs were prospectively recruited. Overall, there was no statistically significant difference between CSF-CK collected from the CM or lumbar cisterna (P = .31). When evaluated by neurolocalization, CSF-CK was different between sites in dogs with thoracolumbar myelopathy (P = .024), but not in dogs with intracranial or cervical neurolocalization (P = .93). All dogs with thoracolumbar myelopathy had equivocal or higher CK levels at the lumbar collection site compared with levels at the CM collection site. CONCLUSIONS: Cerebrospinal fluid CK values differed depending on the CSF site collection, especially in dogs with thoracolumbar myelopathy. In dogs with thoracolumbar myelopathy, CSF-CK was likely to be higher when CSF was taken from the lumbar cisterna compared with the CM cisterna. Collecting CSF from the thoracolumbar site could provide better prognostic information than if collected at the CM collection site.

Biological behaviour and ezrin expression in canine rhabdomyosarcomas: 25 cases (1990-2012).

There are few published reports of canine rhabdomyosarcomas. In human paediatrics, rhabdomyosarcomas account for 5%-10% of all tumours and >50% of soft tissue sarcomas. They have an aggressive biologic behaviour; most patients develop diffuse metastatic disease. Ezrin, a cytoskeleton linker protein, has been correlated with metastasis in a number of tumours, including rhabdomyosarcomas. The goal of this study was to describe dogs with non-urinary rhabdomyosarcomas including clinical findings, ezrin expression and outcome. Twenty-five dogs with rhabdomyosarcomas were identified from two institutions' databases. Signalment, primary tumour location, cytologic and histologic findings, metastatic sites, treatments, survival time and necropsy results were recorded. Immunohistochemical staining for ezrin expression was performed on archived samples; cellular localization of ezrin was characterized. The mean and median age of all patients was 4.3 and 2 years, respectively. Subcutaneous and retrobulbar/orbital were the most common primary tumour locations. Sixteen dogs had metastatic disease at diagnosis. Three dogs presented with diffuse disease where a primary tumour could not be identified. A round cell tumour was the initial diagnosis in 32% of cases, and 76% of cases required immunohistochemistry to establish the diagnosis. The median survival was 10 days. Twenty-one cases had archived samples available for ezrin staining; all but one was positive and exhibited both membranous and cytoplasmic localization. Rhabdomyosarcomas occur in young dogs, may have a round cell appearance, and exhibit aggressive biologic behaviour. Given ezrin's defined role in metastasis, its observed expression in the tumours in this study suggest its possible role in canine rhabdomyosarcoma's aggressive biologic behaviour.

Comparison of cerebellomedullary and lumbar cerebrospinal fluid analysis in dogs with neurological disease.

BACKGROUND: Cerebrospinal fluid (CSF) analysis aids in categorizing underlying disease processes in patients with neurologic disease. Convention suggests that CSF should be collected caudal to the lesion. However, little evidence exists to justify this assertion. HYPOTHESIS/OBJECTIVES: Evaluate the clinicopathologic differences between CSF collected from the cerebellomedullary (CM) and lumbar cisterns in dogs presented for evaluation of neurologic disease. ANIMALS: Fifty-one client-owned dogs undergoing magnetic resonance imaging (MRI) and CSF collection for investigation of neurologic disease. METHODS: Cerebrospinal fluid was prospectively collected from the CM and lumbar cisterns in all patients. The total protein (TP) concentration, red blood cell (RBC) count, and total nucleated cell count (TNCC) were analyzed within 30 minutes of collection. Results and cytology findings were interpreted by a single pathologist. RESULTS: Fifty-one paired samples were collected. The TNCC (P < .001), RBC (P < .001), and TP (P < .001) were different between collection sites. When grouped by neurolocalization, TP (intracranial, P < .001; cervical, P < .001; thoracolumbar, P < .001) and RBC (intracranial, P < .001; cervical, P ≤ .002; thoracolumbar, P = .006) counts were significantly different. The TNCC was significantly different in the cervical (P = .04) and thoracolumbar localizations (P = .004) but not for intracranial (P = .30) localizations. The pathologist's interpretation differed between sites in 66.7% of the cases (34/51). CONCLUSIONS: In dogs with lesions that neurolocalized to the brain or cervical spinal cord, there may be clinical benefit in collecting fluid from both the CM and lumbar cisterns. In dogs with thoracolumbar myelopathy, CSF collected from the CM cistern may not be representative of the underlying disease process.

Overexpression of prostate specific membrane antigen by canine hemangiosarcoma cells provides opportunity for the molecular detection of disease burdens within hemorrhagic body cavity effusions.

BACKGROUND: Canine hemangiosarcoma (cHSA) is a highly metastatic mesenchymal cancer that disseminates by hematogenous and direct implantation routes. Therapies for cHSA are generally ineffective, in part due to advanced clinical disease stage at the time of diagnosis. The validation of conventional molecular methods for detecting novel biomarkers preferentially expressed by cHSA could lead to more timely diagnosis, earlier therapeutic interventions, and improved outcomes. In humans, prostate-specific membrane antigen (PSMA) is a transmembrane protein overexpressed by prostate carcinoma and tumor-associated endothelium of various solid cancer histologies. Importantly, the preferential overexpression of PSMA by certain cancers has been leveraged for the development of diagnostic molecular imaging reagents and targeted therapeutics. Recently, PSMA has been qualitatively demonstrated to be expressed in cHSA cell lines, however, quantitative PSMA expressions and the potential utility of PSMA transcript identification in biologic fluids to support the presence of microscopic cHSA burden has not been reported. Therefore, this study sought to characterize the differential quantitative expressions of PSMA between cHSA and non-malignant tissues, and to determine the potential diagnostic utility of PCR-generated PSMA amplicons as a surrogate of rare cHSA cells dwelling within peritoneal and pericardial cavities. METHODS: Quantitative gene and protein expressions for PSMA were compared between one normal endothelial and six cHSA cell lines by RT-PCR, western blot analysis, and fluorescent microscopy. Additionally, gene and protein expressions of PSMA in normal canine tissues were characterized. Graded expressions of PSMA were determined in spontaneously-arising cHSA tumor samples and the feasibility of qualitative PCR as a molecular diagnostic to detect PSMA transcripts in whole blood from healthy dogs and hemorrhagic effusions from cHSA-bearing dogs were evaluated. RESULTS: PSMA gene and protein expressions were elevated (up to 6-fold) in cHSA cells compared with non-malignant endothelium. By immunohistochemistry, protein expressions of PSMA were detectable in all cHSA tissue samples evaluated. As predicted by human protein atlas data, PSMA's expression was comparably identified at substantial levels in select normal canine tissues including kidney, liver, and intestine. In young healthy pet dogs, PSMA amplicons could not be identified in circulating whole blood yet were detectable in hemorrhagic effusions collected from pet dogs with confirmed cHSA or PSMA-expressing cancer. CONCLUSIONS: PSMA is quantitatively overexpressed in cHSA compared to normal endothelium, but its protein expression is not restricted to only cHSA tumor tissues, as specific visceral organs also substantively express PSMA. Optimized qualitative PCR methods failed to amplify PSMA amplicons sufficiently for visible detection in circulating whole blood derived from healthy young dogs, yet PSMA transcripts were readily identifiable in hemorrhagic effusions collected from pet dogs with histologically confirmed cHSA or PSMA-expressing cancer. While preliminary, findings derived from a limited cohort of normal and diseased pet dogs provocatively raise the potential value of PSMA amplicon detection as an ancillary molecular diagnostic test for supporting the presence of microscopic cHSA disease burden within hemorrhagic body cavity effusions.

The ACVP/STP Coalition for Veterinary Pathology Fellows Celebrates 32 New Training Positions.

Since its creation in 2004, the Coalition for Veterinary Pathology Fellows, a partnership between the American College of Veterinary Pathologists and the Society of Toxicologic Pathology, has established 32 new training positions backed by US$7.4 million in financial support from private sponsors.

Cytology of Bone.

Cytology of bone is a useful diagnostic tool. Aspiration of lytic or proliferative lesions can assist with the diagnosis of inflammatory or neoplastic processes. Bacterial, fungal, and protozoal organisms can result in significant osteomyelitis, and these organisms can be identified on cytology. Neoplasms of bone including primary bone tumors such as osteosarcoma, chondrosarcoma, fibrosarcoma, synovial cell sarcoma, and histiocytic sarcoma and tumors of bone marrow including plasma cell neoplasia and lymphoma and metastatic neoplasia can result in significant bone lysis or proliferation and can be diagnosed effectively with cytology.

SYSTEMIC BLASTOMYCOSIS IN A CAPTIVE RED RUFFED LEMUR (VARECIA RUBRA).

A 5-yr-old, intact male red ruffed lemur ( Varecia rubra ) presented for evaluation as the result of a 1-wk history of lethargy and hyporexia. Physical examination findings included thin body condition, muffled heart sounds, harsh lung sounds, and liquid brown diarrhea. Complete blood count and serum biochemistry showed an inflammatory leukogram, mild hyponatremia, and mild hypochloremia. Orthogonal trunk radiographs revealed a severe alveolar pattern in the right cranial lung lobes with cardiac silhouette effacement. Thoracic ultrasound confirmed a large, hypoechoic mass in the right lung lobes. Fine-needle aspiration of the lung mass and cytology revealed fungal yeast organisms, consistent with Blastomyces dermatitidis. Blastomyces Quantitative EIA Test on urine was positive. Postmortem examination confirmed systemic blastomycosis involving the lung, tracheobronchial lymph nodes, spleen, kidney, liver, cerebrum, and eye. To the authors' knowledge, this is the first report of blastomycosis in a prosimian species.