Immunohistochemical differentiation of reactive from malignant mesothelium as a diagnostic aid in canine pericardial disease

Objectives

To develop a provisional immunohistochemistry panel for distinguishing reactive pericardium, atypical mesothelial proliferation and mesothelioma in dogs.

Materials and Methods

Archived pericardial biopsies were subject to haematoxylin and eosin staining, immunohistochemistry for cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Samples were scored for intensity and number of cells stained.

Results

Ten biopsies of reactive mesothelium, 17 of atypical mesothelial proliferation, 26 of mesothelioma and five of normal pericardium were identified on the basis of haematoxylin and eosin staining. Cytokeratin and vimentin were expressed in all biopsies, confirming mesothelial origin. Normal pericardial samples had the lowest scores for insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Mesothelioma and atypical proliferative samples were similar to each other, with higher scores for insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 than the reactive samples. Desmin staining was variable. Insulin‐like growth factor II mRNA‐binding protein 3 was the best to distinguish between disease groups.

Clinical Significance

An immunohistochemistry panel of cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 could provide superior information compared with haematoxylin and eosin staining alone in the diagnosis of cases of mesothelial proliferation in canine pericardium, but further validation is warranted.     

The influence of rapid growth on sodium salicylate pharmacokinetics in male turkeys

The aim of this study was to assess the influence of growth on the pharmacokinetics of sodium salicylate (SS) in male turkeys. SS was administered intravenously at a dose of 50 mg/kg. Plasma drug concentrations were assessed by high‐performance liquid chromatography, and pharmacokinetic parameters were calculated by noncompartmental analysis. As the age increased from 6 to 13 weeks (body weight increase from 2.35 to 9.43 kg), median body clearance decreased from 1.34 to 0.87 ml/min/kg. This caused a significant increase in the median mean residence time from 3.42 to 4.44 hr. Elimination phase proved to be biphasic and two elimination half‐lives (T_1/2el) were distinguished. Whereas T_1/2el1 was found to increase with age by 128%, T_1/2el2 represented a later but faster and less age‐dependent phase of elimination (increase by 56% in the respective groups). Volume of distribution decreased with age. These effects may lead to different therapeutic response to SS in turkeys of different age and body weights.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

COMPARISON OF SONOGRAPHIC FEATURES OF BENIGN AND NEOPLASTIC DEEP LYMPH NODES IN DOGS

The differentiation of benign vs. neoplastic lymph nodes impacts patient management. Specific sonographic features are typically considered when assessing lymph nodes in dogs. However, the usefulness of these criteria in distinguishing benign vs. malignant lymph nodes remains largely unknown, especially for deep lymph nodes. Our aim was to compare sonographic features in benign and neoplastic deep lymph nodes with the hope of identifying predictive criteria. Thirty‐one deep lymph nodes (16 mesenteric, 10 medial iliac, three hepatic, one sternal, and one cranial mediastinal) in 31 dogs were examined prospectively with B‐mode and Color flow Doppler. Lymph nodes were aspirated using ultrasound‐guidance and final diagnosis were established based on cytologic and/or histopathologic interpretation. Prevalence of each sonographic feature and combinations of two features was calculated for each group and compared using a χ²‐test or Student's t‐test for unequal variances. Ten lymph nodes were benign (hyperplastic and/or inflammatory) and 21 were neoplastic. All were hypoechoic, except for one neoplastic lymph node. Maximal short‐axis diameter (P=0.0006) and long‐axis diameter (P=0.01), and SA/LA ratio (P=0.008) were increased significantly for neoplastic (2.8, 5.5 cm, and 0.50, respectively) vs. benign (1.2, 3.8 cm, and 0.34, respectively) lymph nodes. The prevalence of other features was similar between groups. Doppler evaluation was possible in 77% of lymph nodes, but there was no significant difference between groups. When any two ultrasound features were combined, the only difference between benign and neoplastic lymph nodes was for the combination of contour regularity and appearance of the perinodal fat (P=0.03).     

A Simple and Rapid PCR‐Based Method for Ostrich Sexing Using Micro Amounts of DNA

The aim of the present study is to identify ostrich sex by using polymerase chain reaction (PCR) on micro amounts of DNA from blood, bloodstain and feathers. Sixteen male and 18 female ostriches were used as test individuals. Genomic DNA as a template was extracted by the Chelex method. Ostrsex‐P1 and P2 primers were designed to perform PCR amplification on the template. PCR products were checked using agarose gel electrophoresis with ethidium bromide staining and ostrich sex was determined directly by the bands shown on the gel. The results demonstrate that ostrich sex can be determined by the extraction of DNA from as little as 0.0125 μl blood using Chelex, whereby the use of large amounts of organic solvents such as phenol and chloroform are unnecessary. In addition, it is possible to identify ostrich sex using micro amounts of DNA extracted from bloodstains and/or feathers. The use of feathers particularly avoids unwanted sampling problems such as the difficulty of collecting ostrich blood, the stress to the ostrich caused by bleeding, and the demand for a lot of manpower for ostrich restraint.     

Serum biomarker profiling in cancer studies: a question of standardisation?

Companion animals are exposed to similar environmental conditions and carcinogens as humans. In some animal cancers, there also appears to be the same genetic changes associated as in humans. However, little work has been carried out in cancer biomarker identification in animals. The recent dramatic advances in molecular medicine, genomics, proteomics and translational research will allow biomarker identification, which may provide the best strategies for veterinarians and clinicians to combat disease by early diagnosis and administration of effective treatments. Proteomics may have important applications in cancer diagnosis, prognosis and predictive clinical outcome that could directly change clinical practice by affecting critical elemen‐ts of care and management. This review summarizes the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality. In this review, we will discuss the available proteomic technologies and their limitations, and highlight the key areas of research and how they have been used to discover cancer biomarkers. The principles described here are equally applicable to human and animal disease, but implementation of ‘omic’ technologies requires stringent guidelines for collection of clinical material, the application of analytical techniques and interpretation of the data.     

Association between weight change during initial chemotherapy and clinical outcome in dogs with multicentric lymphoma

The majority of the known prognostic factors in dogs with lymphoma have been evaluated before treatment commences or at the time of diagnosis. Prognostic factors evaluated during the initial phase of treatment are less described but may provide important clinical information. In this retrospective study, 82 canine lymphoma patients were categorized according to the weight change between diagnosis and after 5 weeks of chemotherapy. Dogs that gained greater than 5% or lost greater than 5% of initial body weight were categorized as increased‐ or decreased‐weight groups, respectively. Those in which weight changed less than 5% were categorized as the maintained‐weight group. The median progression‐free survival (PFS) in the increased‐weight group, maintained‐weight group and decreased‐weight group was 226, 256 and 129 days, respectively. The decreased‐weight group had significantly shorter PFS than the increased and maintained groups (P = .023, P = .003, respectively). The median survival time (ST) in the increased‐weight group, maintained‐weight group and decreased‐weight group was 320, 339 and 222 days, respectively. There was no significant difference in ST among the three groups (P = .128). In Cox‐regression results, weight change group and initial body weight were significant risk factors associated to PFS (P = .007, P = .001, respectively) while only patient's initial body weight was a significant risk factor to ST (P = .013). In conclusion, evaluation of initial body weight and weight changes over time can provide valuable information regarding PFS and ST in dogs with multicentric lymphoma.