Lisbeth Rem Jessen

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 C-reactive protein as an early marker of inflammatory activity in canine type II immune-mediated polyarthritis: case report.

BackgroundMonitoring systemic inflammatory activity during steroid therapy of canine immune-mediated polyarthritis (IMPA) is difficult and mainly relies on clinical signs.Case presentationCanine serum C-reactive protein (CRP) was measured serially and blinded during a 27-week follow-up period of a case of Anaplasma phagocytophilia induced type II immune-mediated polyarthritis.ConclusionWBC was, as expected, observed not to reflect the inflammatory activity during steroid treatment in a clinical useful manner, whereas, CRP is suggested a valuable unbiased marker of inflammatory activity during steroid treatment in this case.

In vitro heparinization of canine whole blood with low molecular weight heparin (dalteparin) significantly and dose-dependently prolongs heparinase-modified tissue factor-activated thromboelastography parameters and prothrombinase-induced clotting time

BACKGROUND Low-molecular-weight heparin (LMWH) is being used increasingly in veterinary medicine for both treatment and prophylaxis of thromboembolic disease, but no predictable patient-side method exists to monitor its effect. OBJECTIVES The aim of this study was to evaluate thromboelastography (TEG) and prothombinase-induced clotting time (PiCT) assays for detecting hemostatic alterations following in vitro heparinization of canine whole blood with dalteparin (Fragmin). METHODS Citrated whole-blood samples were collected from 7 clinically healthy dogs. Dalteparin was added at concentrations of 0, 0.156, 0.625, 1.25, and 2.5 U/mL of whole blood. TEG was performed using heparinase cups with tissue factor (TF, 1:50,000) and kaolin as activators. Reaction time (R), clotting time (K), angle (alpha), and maximum amplitude (MA) were recorded. PiCT and anti-FXa activity were measured in plasma. RESULTS With TF, increasing concentrations of dalteparin significantly prolonged R and K and significantly decreased alpha and MA. K, alpha, and MA ratios were significantly different from baseline at all dalteparin concentrations and R was significantly different from baseline at concentrations of 0.625, 1.25, and 2.5 U/mL. With kaolin, only R was significantly different from baseline at dalteparin concentrations of 0.625 and 2.5 U/mL. PiCT detected dalteparin concentrations < or = 0.625 U/mL, with a good linear correlation (r(2)=.96, P<.0001). CONCLUSION These results suggest that TF-activated TEG and PiCT assays should be further evaluated as promising new methods for evaluating the effect of LMWH, using doses in the recommended clinical range and prospective clinical studies.

Endogenous fibrinolytic potential in tissue-plasminogen activator-modified thromboelastography analysis is significantly decreased in dogs suffering from diseases predisposing to thrombosis

BACKGROUND In people, studies have shown that resistance to fibrinolysis could be a contributing factor to thrombosis. Tissue-plasminogen-activated (t-PA) thromboelastography (TEG) has been used to evaluate endogenous fibrinolytic potential. In dogs, TEG has been used for the diagnosis of various hemostatic disorders, but studies evaluating fibrinolysis are limited. Investigations into the potential of t-PA-modified TEG to monitor endogenous fibrinolytic potential are lacking in both healthy dogs and dogs with diseases predisposing to development of thrombosis. OBJECTIVES The aim of this study was to compare 3 t-PA-modified TEG assays and compare the endogenous fibrinolytic potential in dogs suffering from diseases associated with thrombosis with a group of healthy dogs. METHODS Three different TEG assays, such as native, tissue factor-activated, and kaolin-activated, were modified with t-PA and used to compare whole blood samples from 16 healthy control dogs and 20 diseased dogs. RESULTS Thromboelastography lysis variables were significantly affected by addition of t-PA in all 3 assays. Lysis results in diseased dogs were comparable to those in healthy dogs prior to addition of t-PA. After addition of t-PA, lysis results were significantly decreased in the diseased group compared with healthy dogs. The lowest median lysis levels were found in dogs with systemic inflammation and protein-losing disorders. CONCLUSION Addition of t-PA activates fibrinolysis in TEG of blood from both healthy dogs and dogs with diseases predisposing to thrombosis. The significantly decreased fibrinolysis in diseased dogs suggests that this may be a potential prothrombotic risk factor in dogs.

Point of care susceptibility testing in primary care - does it lead to a more appropriate prescription of antibiotics in patients with uncomplicated urinary tract infections? Protocol for a randomized controlled trial

BackgroundUrinary tract infection (UTI) is a common infection in primary care and is the second leading reason for prescription of antibiotics in Denmark. The diagnosis is often based on symptoms and urine dip-stick, which has limited validity, causing the risk of unnecessary antibiotic prescription. Additionally, with increasing antibiotic resistance, the risk of choosing an antibiotic to which an infecting pathogen is resistant is rising. Combined point-of-care-tests (POCT) for urine culture and susceptibility testing have been developed and validated for primary care, and performing such a test in all patients with suspected UTI in primary care seems rational in order to reduce the use of inappropriate antibiotics. However, the clinical effect of the culture and susceptibility test has not yet been investigated. This study aims to investigate whether POCT urine culture and susceptibility testing decreases the inappropriate use of antibiotics and leads to faster patient recovery.Methods/designRandomized controlled open label trial of two diagnostic approaches. 750 patients with symptoms of uncomplicated UTI, consecutively contacting their general practitioner (GP), randomized to either POCT urine culture and susceptibility testing and targeted treatment or POCT urine culture without susceptibility testing and empirical treatment. Treatment is started when the POCT is read. The two groups are compared with regard to appropriate choice of antibiotics, clinical remission, and microbiological cure rates.DiscussionThe results of this study may provide important evidence to recommend POCT culture and susceptibility testing in all patients with suspected uncomplicated UTI. This could become an additional strategy to fight antibiotic resistance.Trial registrationClinicalTrials.gov NCT02323087.

Effect of antibiotic treatment in canine and feline urinary tract infections: a systematic review.

Urinary tract infection (UTI) is a major reason for antibiotic prescription in small animal practice. Optimal antibiotic treatment strategies have not been established for veterinary species, especially when considering duration of treatment, which is often considerably longer than for human patients with UTI. The aims of this study were (1) to identify and assess evidence related to the efficacy of antibiotic treatment in canine and feline UTIs; and (2) to compare the efficacy of short (<5 days) and standard (≥7 days) duration of antibiotic treatment for canine uncomplicated UTI. An electronic literature search was conducted for publications to 1 May 2014. Fourteen peer-reviewed prospective and controlled studies were retrieved, 10 of which evaluated antibiotic treatment in dogs and four in cats. Of the 14 studies, seven were clinical trials and five of those were randomised controlled trials. Most (12/14) studies were not considered to contribute sufficient evidence to evaluate treatment strategies. There were no clinical studies examining the effect of duration of the same drug. Of the short duration regimens evaluated, the efficacy of 3 day antibiotic therapy with trimethoprim-sulphonamide (females only) or high-dose enrofloxacin in dogs with uncomplicated UTIs was supported by fair evidence, as these treatment strategies were non-inferior to medium duration (10-14 days) therapy with β-lactam antimicrobials. In conclusion, there is little published evidence relating to antibiotic treatment of UTIs in dogs and cats. Well-designed clinical trials focusing on the duration of treatment are warranted to create evidence-based treatment protocols.

Diagnosis and treatment of platelet hyperactivity in relation to thrombosis in dogs and cats

OBJECTIVE To review the mechanisms of platelet activation and options for diagnosing and treating platelet hyperactivity in relation to thrombosis in dogs and cats. DATA SOURCES Prospective, retrospective, and review articles, as well as textbook chapters in both human and veterinary medicine. Articles were primarily, but not exclusively, retrieved via Medline. HUMAN DATA SYNTHESIS In people, platelets are known to play a key role in the development of arterial thrombosis in numerous disease states and antiplatelet drugs are the cornerstone in the treatment of acute events and for prevention in patients at risk. For many years, aspirin was used as the sole antiplatelet drug in people, but the introduction of adenosine diphosphate receptor antagonists and integrin α(IIb) β(3) inhibitors has significantly improved outcome in selective groups of patients. VETERINARY DATA SYNTHESIS The understanding of platelet activation in disease states has increased dramatically over the past decade. Simultaneously, a host of new methods for evaluating platelet function have been developed, which enable primarily researchers, but also clinicians to monitor the activity of platelets. Many of these methods have been validated for research purposes, but few have found their way to the clinics. Not a single correctly randomized clinical trial has been carried out with any antiplatelet drug for any indication in dogs or cats, and consequently, treatment is empiric and largely based on expert opinion or data from experimental studies. CONCLUSIONS The pathogenesis of thromboembolic disease is complex and multifactorial and the role of hyperactive platelets in this etiology remains to be clarified in most of the diseases associated with thrombosis in dogs and cats. Until efficacy data from well-designed studies are available, antithrombotic therapy should consist of close monitoring, good supportive care, and judicious empirical use of antiplatelet agents.Objective To review the mechanisms of platelet activation and options for diagnosing and treating platelet hyperactivity in relation to thrombosis in dogs and cats. Data Sources Prospective, retrospective, and review articles, as well as textbook chapters in both human and veterinary medicine. Articles were primarily, but not exclusively, retrieved via Medline. Human Data Synthesis In people, platelets are known to play a key role in the development of arterial thrombosis in numerous disease states and antiplatelet drugs are the cornerstone in the treatment of acute events and for prevention in patients at risk. For many years, aspirin was used as the sole antiplatelet drug in people, but the introduction of adenosine diphosphate receptor antagonists and integrin αIIbβ3 inhibitors has significantly improved outcome in selective groups of patients. Veterinary Data Synthesis The understanding of platelet activation in disease states has increased dramatically over the past decade. Simultaneously, a host of new methods for evaluating platelet function have been developed, which enable primarily researchers, but also clinicians to monitor the activity of platelets. Many of these methods have been validated for research purposes, but few have found their way to the clinics. Not a single correctly randomized clinical trial has been carried out with any antiplatelet drug for any indication in dogs or cats, and consequently, treatment is empiric and largely based on expert opinion or data from experimental studies. Conclusions The pathogenesis of thromboembolic disease is complex and multifactorial and the role of hyperactive platelets in this etiology remains to be clarified in most of the diseases associated with thrombosis in dogs and cats. Until efficacy data from well-designed studies are available, antithrombotic therapy should consist of close monitoring, good supportive care, and judicious empirical use of antiplatelet agents.

Investigation of a screening programme and the possible identification of biomarkers for early disseminated histiocytic sarcoma in Bernese Mountain dogs

The aim of the study was to construct a screening programme for disseminated histiocytic sarcoma (DHS) in Bernese Mountain dogs using diagnostic imaging and blood analysis and evaluate blood borne biomarkers as early disease detection biomarkers. Healthy Bernese Mountain dogs were screened on four occasions in an attempt to detect early disease. Eleven blood borne biomarkers were examined for their worth as early tumour biomarkers. During 2.5 years, five dogs with early DHS were identified; four of these by diagnostic imaging. No dogs developed symptomatic DHS without being detected within 6 months of the screening programme. Only serum ferritin showed potential as a blood borne marker of the disease. Median survival times for the dogs with early DHS were 226 days. Screening programmes every 6 months for Bernese Mountain dogs over 4 years of age including diagnostic imaging and ferritin measurements may identify early DHS.

Thrombin-activatable fibrinolysis inhibitor activity in healthy and diseased dogs

BACKGROUND In people, increased thrombin-activatable fibrinolysis inhibitor (TAFI) antigen has been associated with increased risk of thrombosis, and decreased TAFI may contribute to bleeding diathesis. TAFI activity in dogs has been described in experimental models, but not in dogs with spontaneous disease. OBJECTIVE The aim of this study was to compare TAFI activity in healthy dogs with TAFI activity in dogs with spontaneous disease. METHODS Plasma samples from 20 clinically healthy Beagles and from 35 dogs with various diseases were analyzed using a commercial chromogenic assay that measured TAFI activity relative to activity in standardized pooled human plasma. RESULTS Median TAFI activity for the 20 Beagles was 46.1% (range 32.2-70.8%) compared with 62.6% (29.1-250%) for the 35 diseased dogs, and 14/35 (40%) had TAFI activities >the upper limit for controls. The highest individual activities (>225%) were in 3 dogs with malignant neoplasms and 1 dog with thrombocytopenia. For data grouped by diagnosis, median TAFI activity was 61.7% for benign neoplasia (n=5), 64.9% for malignant neoplasia (n=8), 75.5% for Angiostrongylus vasorum infection (n=4), 68.8% for bacterial sepsis (n=7), and 58.7% for miscellaneous diseases (n=11). Compared with TAFI activity in control dogs, median TAFI activity was significantly increased only in the group of dogs with bacterial sepsis. CONCLUSION Bacterial sepsis was associated with significantly increased TAFI activity, and individual dogs with increased TAFI activities were found in all disease groups. The role of TAFI in the pathogenesis of hemostatic disorders in dogs and its value as a prognostic indicator deserve further investigation.

Evaluation of different sampling methods and criteria for diagnosing canine urinary tract infection by quantitative bacterial culture

The use of voided urine specimens for bacteriological culture in dogs is discouraged because contamination from external genitalia could lead to misinterpretation of laboratory results. Quantitative culturing and defining significant bacteriuria could increase the usefulness of voided specimens. However, limited evidence exists for the cut-offs currently recommended. The aim of this study was to evaluate the accuracy of current veterinary cut-off values for significant bacteriuria in voided canine urine. A secondary aim was to investigate if accuracy improved when applying qualitative criteria used in humans. Paired urine specimens were collected by both cystocentesis and voiding, and quantitative bacteriological cultures were performed within the same day. Cystocentesis was used as the reference standard with a cut-off for significant bacteriuria of ≥1000 colony forming units (CFU)/mL. Voided specimens were compared to cystocentesis using: (1) the veterinary cut-off of ≥100,000 CFU/mL; and (2) various cut-offs depending on qualitative criteria (sex, clinical signs and complicating factors), adapted from human guidelines. Ninety-four dogs with suspected urinary tract infection (UTI) were included for analysis. The veterinary cut-off yielded an accuracy of 94% with a sensitivity and specificity of 94% (95% confidence intervals [CI] 0.81, 0.99) and 94% (95% CI 0.86, 0.98), respectively. Applying the human guidelines did not improve overall accuracy (89%), and yielded a sensitivity and specificity of 97% (95% CI 0.86, 1.00) and 86% (95% CI 0.77, 0.92), respectively. The veterinary cut-off value of ≥100,000 CFU/mL for voided urine is appropriate for determining significant bacteriuria in the majority of dogs with suspected UTI if specimens are refrigerated and cultured on the day of collection.