A. L. Jensen

Thromboelastographic Evaluation of Hemostatic Function in Dogs with Disseminated Intravascular Coagulation

BACKGROUND There is considerable variation in the coagulation profile of dogs with disseminated intravascular coagulation (DIC), making it difficult to assess overall hemostatic function. OBJECTIVES To characterize the overall hemostatic state in dogs with DIC, by use of tissue factor-activated thromboelastography (TF-TEG), and to determine whether there is an association between hemostasis and outcome. ANIMALS 50 dogs with DIC. METHODS Dogs admitted to the intensive care units, with an underlying disease known to predispose to DIC, were prospectively assessed with TF-TEG. Citrated blood samples were collected daily during hospitalization and an extended coagulation panel and TF-TEG were performed. Diagnosis of DIC was based on expert opinion. RESULTS Hemostatic dysfunction was observed on the TF-TEG profile in 33/50 of the dogs, of which 22/50 were hypercoagulable and 11/50 were hypocoagulable based on the TF-TEG G value alone. There were significant differences in k, alpha, and MA values (P < .0001) among hypo-, normo-, and hypercoagulable dogs. There was a significant difference in case fatality rate between hypo- (64%) and hypercoagulable (32%) dogs (relative risk = 2.38; P= .04). Dogs that died had significantly lower antithrombin activity (P= .03) and higher d-dimer concentration (P= .03) than survivors. CONCLUSIONS The most common overall hemostatic abnormality in dogs diagnosed with DIC was hypercoagulability, and there was significant difference in survival between hyper- and hypocoagulable dogs. The results suggest TF-TEG is valuable in the assessment of hemostatic function in dogs diagnosed with DIC.

Evaluation of human recombinant tissue factor-activated thromboelastography in 49 dogs with neoplasia.

BACKGROUND Abnormal routine coagulation assay results have been reported to be common in veterinary patients with neoplasia, but the overall hemostatic functional state, including hypercoagulability, has not been described. HYPOTHESIS The overall hemostatic functional state, including hypercoagulability, can be assessed in dogs with neoplasia by tissue factor (TF)-activated thromboelastography (TEG). ANIMALS Thirty-six dogs with malignant neoplasia and 13 dogs with benign neoplasia presented to the Small Animal Veterinary Teaching Hospital, The University of Copenhagen, Frederiksberg, Denmark. METHODS Prospective study evaluating the overall hemostatic functional state in dogs with neoplasia by a newly validated TF-activated TEG assay and routine coagulation parameters activated partial thromboplastin time (aPTT), prothrombin time (PT), platelet count, and D-dimer concentration. RESULTS Hemostatic dysfunction was observed in 28/49 (57%) dogs with neoplasia. Twenty-four were dogs with malignant neoplasia, the majority of which 18/36 (50%) were hypercoagulable, whereas 6/36 (17%) were hypocoagulable. All hypocoagulable dogs had metastatic disease. The proportion of dogs with altered hemostasis was significantly different between dogs with malignant and benign neoplasia. CONCLUSIONS AND CLINICAL IMPORTANCE TF-activated TEG detected hypercoagulable and hypocoagulable states in this population of dogs with neoplasia. The most common hemostatic abnormality in dogs with malignant neoplasia was hypercoagulability. These findings suggest that this novel hemostatic function test may be of value as a cage side method for the assessment of overall hemostatic function in dogs with cancer, including the detection of both hyper- and hypocoagulable states as well as mixed disorders.

Use of serum amyloid A and other acute phase reactants to monitor the inflammatory response after castration in horses: a field study

REASONS FOR PERFORMING THE STUDY Early recognition of excessive inflammation and infectious complications after surgery, leading to early institution of therapy, reduces post operative discomfort and facilitates recovery. Because serum amyloid A (SAA) is a highly sensitive marker of inflammation, measurements of SAA and other acute phase reactants in the equine surgical patient may be valuable in assisting clinical assessment of post operative inflammation. OBJECTIVES To investigate changes in inflammatory markers after castration and to correlate levels of acute phase reactants with clinical severity of inflammation after castration. METHODS Leucocyte numbers and blood levels of iron, SAA and fibrinogen were determined before castration and on Days 3 and 8 post operatively in 2 groups of horses; Group 1 (n = 11) had mild post operative inflammation and an uncomplicated recovery and Group 2 (n = 7) had local clinical signs of moderate to severe inflammation. RESULTS Both groups had elevated serum SAA levels at Day 3 post operatively. In Group 1 concentrations had returned to preoperative levels by Day 8, whereas in Group 2 concentrations remained elevated. Plasma fibrinogen concentrations in serum increased to equal levels in both groups and stayed elevated throughout the study period. Serum iron concentrations of Group 1 did not change in response to castration, whereas concentrations in Group 2 decreased below preoperative levels on Day 8. Leucocyte numbers remained unchanged during the post operative period in both groups. CONCLUSIONS Serum SAA and iron profiles reflected the course of inflammation and their levels correlated with the clinical severity of inflammation. In contrast, fever and changes in leucocyte numbers, which are usually considered to be hallmarks of inflammation and infection, were not useful for monitoring post operative recovery. POTENTIAL RELEVANCE Measurements of SAA and iron may improve post operative monitoring. As sustained inflammation may indicate that the surgical wound has become infected, SAA and iron measurements may facilitate early recognition and hence early treatment of infection.

Mercury contamination in spotted seatrout, Cynoscion nebulosus: An assessment of liver, kidney, blood, and nervous system health

Marine fishes in South Florida (Florida Keys-Florida Bay-Everglades region) accumulate higher concentrations of mercury (Hg) in their tissues than similar fishes from other areas of the southeastern U.S., though it is not known whether these elevated levels affect fish health. In this study, we used quantifiable pathological and biochemical indicators to explore Hg-associated differences in marine fish from South Florida, where Hg contamination is high, and from Indian River Lagoon, Florida, which served as a reference area. Hg concentrations in all tissues of mature spotted seatrout (Cynoscion nebulosus) from South Florida were significantly higher than those from Indian River Lagoon and were within the threshold range of those in studies where effects of Hg exposure have been observed. The distribution of Hg among tissues followed the same trend in both areas, with the greatest concentration in kidney tissue, followed by liver, muscle, brain, gonad, and red blood cells. Blood-plasma biochemistry showed that concentrations of iron, inorganic phosphate, lactate dehydrogenase, and aspartate aminotransferase were significantly less in South Florida. Also, fructosamine and alkaline phosphatase were significantly less in South Florida. Liver histology revealed that pyknosis/necrosis, interstitial inflammation, and bile duct hyperplasia were found only in seatrout from South Florida, and steatosis/glycogen was more frequently found in Indian River Lagoon specimens. In renal tissue, interstitial inflammation, glomerular dilatation and thickening, and tubular degeneration and necrosis were more frequently found in South Florida specimens. Changes in the liver cytoskeleton and morphology may explain some of the differences in blood parameters between study areas. Neurochemical analyses showed that brain N-methyl-d-aspartic acid (NMDA) receptors (but not those of muscarinic cholinergic receptors, monoamine oxidase, or acetylcholinesterase) were significantly less in fish from South Florida than from Indian River Lagoon. These findings provide compelling evidence that elevated Hg could cause quantifiable pathological and biochemical changes that might influence the health of spotted seatrout and could also affect other marine fish species.

IL-1β, IL-6, KC and MCP-1 are elevated in synovial fluid from haemophilic mice with experimentally induced haemarthrosis

Summary.  The hallmark of haemophilia is the joint morbidity resulting from haemarthrosis that accounts for the majority of the bleeds. The exact mechanisms underlying changes are not fully elucidated. Cytokines are speculated to be involved in the progression and in vitro studies have confirmed the presence of elevated levels of cytokines in synovial tissue and cartilage from patients with haemophilic synovitis. In this study, the presence of selected cytokines in synovial fluid from haemophilia A mice with experimentally induced haemarthroses treated with rFVIII, rFVIIa and an rFVIIa analogue were investigated. Ten cytokines previously shown to be involved in arthritic syndromes were evaluated. Interleukin (IL)‐1β, IL‐2, IL‐4, IL‐6, IL‐10, IL‐17, Tumor Necrosis Factor‐alpha (TNF‐ α), keratinocyte‐derived chemokine (KC), Regulated upon Activation, Normal T cell Expressed and Secreted (RANTES) and monocyte chemotactic protein‐1 (MCP‐1) were included. In this article, we demonstrate, for the first time, that bleeding in knee joints of haemophilia A mice resulted in correlated increased levels of the pro‐inflammatory cytokines: IL‐1β, IL‐6, KC and the MCP‐1 in synovial fluid. These results suggest an important role of MCP‐1 in the recruitment of monocytes and furthermore that the inflamed synovium releases IL‐1β, IL‐6 and KC, which in turn might contribute to further progression of the inflammatory process.

Critical differences of clinical chemical parameters in blood from dogs

The study concerned the critical difference which may help to judge whether the difference between two consecutive analytical results may be safely ascribed to natural variation or not. To calculate the critical difference of nine canine clinical chemical parameters, blood samples from 20 apparently clinically healthy dogs were collected once weekly for five consecutive weeks. For each of the nine clinical chemical parameters, the total variance of the analytical results was divided into the component of variance between dogs (S2Inter), the component of variance for weeks within dogs (S2Intra) and the component of variance for measurements (S2Anal) using nested analysis of variance. The critical difference was then calculated from S2Intra and S2Anal as 0.22 mu kat litre-1 for alanine aminotransferase, 0.20 mu kat litre-1 for aspartate aminotransferase, 0.34 mu kat litre-1 for alkaline phosphatase, 2.36 mmol litre-1 for urea, 35 mumol litre-1 for creatinine, 2.8 g litre-1 for albumin, 6.3 g litre-2 for serum proteinTotal, 1.49 mmol litre-1 for glucose and 0.84 mmol litre-1 for cholesterolTotal. These critical differences may be used as guidelines to evaluate the difference between two consecutive analytical results of the above parameters. However, the analytical results should not be assessed by the critical differences alone, but should also be compared to the corresponding reference intervals.

Reference interval and critical difference for canine serum fructosamine concentration

The purposes of the study were to obtain a reference interval and to calculate the critical difference between two analytical results for canine serum fructosamine concentration. To obtain a reference interval, the serum fructosamine concentration was measured in blood samples from 29 adult dogs after a 15-h fasting period. To calculate the critical difference, blood samples from 20 apparently clinically healthy dogs were collected once weekly for five consecutive weeks, and the total variance of the analytical results was divided into the component of variance between dogs (Sinter2), the component of variance for weeks within dogs (Sintra2) and the component of variance for measurements (Sanal2), using nested analysis of variance. The critical difference was then calculated fromSintra2 andSanal2.The main conclusions are in summary: The reference interval for canine serum fructosamine concentration is 258.6–343.8 µmol/L, and the critical difference between two consecutive measurements on a week-to-week basis is 32.4 µmol/L. The critical difference may be used as a guideline to indicate potentially important changes in the serum fructosamine concentration, though the analytical results should not be assessed by the critical differences alone, but should also be compared to the corresponding reference intervals.

Serum fructosamine in canine diabetes mellitus. An initial study.

This study reports on a spectrophotometric assay for the determination of serum fructosamine concentration. The assay was evaluated for use in canine serum samples by assessment of the precision, accuracy, detectability and stability of serum fructosamine during storage. To evaluate the diagnostic usefulness of the assay, both the effect of acute changes in blood glucose on serum fructosamine concentration and the serum fructosamine concentration in canine diabetes mellitus and other canine diseases were studied.The main conclusions can be summarized as follows: Determination of canine serum fructosamines may be achieved by a precise and accurate assay with a detection limit well below the serum fructosamine concentration normally found in canine sera. Storage for 5 days at +4°C or +25°C, or for 28 days at −20°C caused no significant change in serum fructosamine concentration. The concentration is not affected by acute changes in blood glucose. In diabetic dogs, serum fructosamine concentration is significantly greater than in dogs with other diseases.

A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5'-diphosphocholine coupled dendrimers.

C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.

Renal lesions in Greenland sledge dogs ( Canis familiaris ) exposed to a natural dietary cocktail of persistent organic pollutants

Persistent organic pollutants such as Polychlorinated biphenyls (PCB) and dichloro diphenyl trichloroethane (DDT) are known to exert various adverse health effects in wildlife mammals. The impact from dietary intake of minke whale (Balaenoptera acutorostrata) blubber high in organohalogen and other chemical contaminants on renal morphology and function was investigated via a controlled study on West Greenland sledge dogs (Canis familiaris). Our results showed significantly higher frequencies of glomerular, tubular, and interstitial lesions in the exposed group. Furthermore, higher urine protein : creatinine ratio and plasma urea levels were found in the exposed group, which indicated a negative impact on kidney function via tubular and glomerular dysfunctions. The lesions were similar to those observed in top predator marine species–such as seals and polar bears–and humans exposed via diet to persistent environmental pollutants. The lesions may have impacts on the overall health of these animals. It is reasonable to suggest that chronic exposure to organic pollutants via a country diet of high trophic level marine organisms may have an impact on renal morphology and function in Arctic apex predators in general as well as Inuit people.