Evaluation of clopidogrel, hypercoagulability, and platelet count in dogs undergoing splenectomy for splenic masses.

Dogs that had splenectomy are predisposed to fatal thrombotic conditions, and thrombocytosis is a risk factor for post-splenectomy hypercoagulability. However, in veterinary medicine, there are no specific therapeutic approaches for managing this hypercoagulability. This study aimed to determine the preventive effect of clopidogrel on post-operative hypercoagulability during the first 2 weeks post-splenectomy in dogs with splenic masses. This study included 12 dogs that had splenectomy. Seven dogs received no treatment (group A), and five were treated with clopidogrel (group B). Clopidogrel was loaded at 10 mg/kg on day 2 and continued at 2 mg/kg until day 14. Blood samples were collected on the day of surgery and 2, 7, and 14 days after splenectomy in both groups. In group B, thromboelastography (TEG) was performed on the same days. In group A, there was significant elevation of platelet counts on days 7 (p = 0.007) and 14 (p = 0.001) compared to day 0. In group B, the platelet counts were significantly elevated on day 7 (p = 0.032) but no significant difference was found on day 14 compared to day 0. Platelet counts on day 14 were significantly higher in group A than in group B (p = 0.03). The lower platelet counts were correlated with alterations in TEG parameters, and no significant differences were found in the K and α-angle values at all postoperative assessment points compared to day 0. Our study suggests that clopidogrel may reduce post-operative thrombocytosis and hypercoagulability in dogs that undergo splenectomy for splenic masses.

Effects of canine influenza infection and DA2PP vaccination on the development of platelet-associated immunoglobulins and platelet counts in dogs.

BACKGROUND: Immune thrombocytopenia (ITP) is commonly associated with platelet-associated immunoglobulins (PAIg). Demonstration of PAIg can help determine etiologies for thrombocytopenia. In humans, ITP and thrombocytopenia have been associated with various vaccinations and influenza infections, respectively. OBJECTIVES: We aimed to evaluate platelet counts and PAIg in research dogs with H3N2 and in research and client-owned dogs routinely vaccinated for distemper, adenovirus-2, parainfluenza, and parvovirus (DA2PP). The hypotheses were that H3N2 infection but not DA2PP vaccination would decrease platelet counts, and neither would result in the detection of PAIg. METHODS: Three pilot studies. Platelet counts and PAIg, measured by direct flow cytometry as %IgG, were evaluated in eight research Beagles following experimental infection with H3N2 (experiment 1), nine research Beagles vaccinated for DA2PP (experiment 2), and thirty client-owned dogs vaccinated for DA2PP (experiment 3). All animals were considered healthy at the start of the experiments. RESULTS: Transient, self-resolving decreases in platelet counts and increases in %IgG occurred following H3N2 infection, and one dog became thrombocytopenic and positive for PAIg. Following DA2PP vaccination, %IgG increased in research and client-owned dogs, but only one dog was considered positive for PAIg with a concurrent increase in platelet count. Mean PAIg increased from baseline in client-owned dogs following vaccination. CONCLUSIONS: Transient PAIg and thrombocytopenia can occur following H3N2 infection, while routine vaccination for DA2PP in this group of dogs was not associated with the development of thrombocytopenia or clinically relevant formation of PAIg.

Prognostic potential of inflammatory markers, oxidative status, thrombocyte indices, and renal biochemical markers in neonatal calf diarrhoea-induced systemic inflammatory response syndrome.

The study aimed to assess the prognostic value of inflammatory markers, indicators of oxidative stress, thrombocyte indices, and renal biochemical markers in neonatal calf diarrhoea (NCD) induced by systemic inflammatory response syndrome (SIRS) upon admission. A prospective, observational, and case-control study was conducted on 56 calves diagnosed with NCD. Mean concentrations of interleukin-6 (IL-6), malondialdehyde (MDA), glutathione (GSH), mean platelet volume (MPV), platelet distribution width (PDW), blood urea nitrogen (BUN), and creatinine (Crea) were measured. Furthermore, the neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were also calculated for SIRS survivors [SIRS (survivor)] and non-survivors [SIRS (non-survivor)] induced by NCD. A prognostic cut-off value for predicting the prognosis of the SIRS's induced by NCD was obtained via receiver operating characteristic (ROC) curve analysis. Upon admission, the SIRS (non-survivor) calves had significantly higher (P < .001) average levels of IL-6, MDA, BUN, Crea, MPV, and PDW compared to the SIRS (survivor) calves and significantly lower (P < .001) average levels of GSH. Despite an apparent increase in the NLR and PLR values of calves diagnosed with SIRS, no significant difference was found between the survival and non-survivor SIRS cases. Positive predictive values (PPVs) for survival were determined as 100 %, 100 %, 80 %, 100 %, 80 %, and 80 %, respectively, using cut-off values of IL-6 (≤259.67 ng/L), MDA (≤2.87 nmol/mL), MPV (≤12.5 fL), PDW (≤34.25 %), BUN (≤168.3 mg/dL), and Crea (≤2.11 mg/dL). The determined threshold values are those obtained upon admission to the hospital. Based on the sensitivity, specificity, and PPVs derived from the ROC analysis, it has been concluded that IL-6, MDA, MPV, PDW, BUN, and Crea are the most relevant biomarkers used for predicting the prognosis of NCD-induced SIRS in calves. Furthermore, it is also noteworthy that IL-6 exhibited the highest effectiveness among all biomarkers.

Platelet count and MCHC as independent prognostic markers for feline mammary carcinomas.

Mammary neoplasms are common in felines species and represent a significant disease for its unfavorable prognosis. Changes in the blood count and serum biochemical profile of these patients have potential as non-invasive prognostic markers prior to mastectomy, however, they are poorly described in literature. In this study univariate and multivariate analyses were performed using these factors to determine the effect of each parameter on the one-year survival time after the surgical procedure in these animals. The median overall survival (OS) and the disease-free survival (DFS) were 365 and 242 days, respectively. In univariate analysis, values within the reference range of monocyte, platelet and creatinine counts were identified as significant prognostic factors for OS and only creatinine was significant for DFS (P < 0.05). In the multivariate analysis, platelets and mean corpuscular hemoglobin concentration (MCHC) remained independent prognostic factors for OS. The results presented suggest that monocytes, platelets and creatinine may be important non-invasive pre-surgical prognostic markers, and that platelet count and MCHC are independent prognostic markers for feline mammary carcinomas (FMC). The correlation between such alterations is of important relevance for veterinary oncology, and prospective studies are needed to validate their clinical use and that platelet count and MCHC are independent prognostic markers for FMC. The results found in this study can also be studied in human medicine, regarding blood markers in human breast cancer (HBC).

Thrombocytosis in 158 cats (2011-2018).

OBJECTIVES: Thrombocytosis is an uncommon hematologic abnormality that is associated with various physiologic, metabolic, inflammatory and neoplastic conditions in people and dogs. Thrombocytosis is not a well-described abnormality in cats. The objective of this study was to classify thrombocytosis in cats based on underlying disease processes and severity, and to compare this with a control population of cats. METHODS: A retrospective study was conducted by reviewing the medical records of cats with increased (>600 × 103/µl; thrombocytosis group) and normal (200-600 × 103/µl; 2:1 age-matched control group) platelet counts between 2011 and 2018. Platelet counts were estimated based on blood smear assessment in all cats. Cats were classified by the severity (mild, moderate or marked) of thrombocytosis. Diagnoses were recorded for all cases, and were classified broadly into either neoplasia, endocrine or inflammatory disease. RESULTS: In total, 158 cats were identified with thrombocytosis, with 315 cats in the control group. Non-neoplastic inflammatory disease was the most common diagnosis in both groups (54.4% in cats with thrombocytosis and 56.2% in controls; P = 0.77); however, gastrointestinal diseases were more common in cats with thrombocytosis (75.6%) when compared with controls (34.5%; P <0.0001). Neoplasia was diagnosed more frequently in cats with thrombocytosis (44.3%) compared with the control group (25.4%; P <0.0001). Round cell tumor was the most common neoplasia diagnosis in both groups, but gastrointestinal and multicentric lymphoma were diagnosed more frequently in cats with thrombocytosis compared with control cats. No association between the severity of thrombocytosis and etiology was identified. CONCLUSIONS AND RELEVANCE: Thrombocytosis in cats is more commonly associated with gastrointestinal, hepatobiliary or immune-mediated diseases when compared with a control population. Neoplasia, especially multicentric and gastrointestinal lymphoma, was more commonly diagnosed in cats with thrombocytosis when compared with control cats.

Lymphopenia predicts reduced survival in canine hepatocellular carcinoma.

Platelet to lymphocyte ratio (PLR) is a prognostic marker in human hepatocellular carcinoma (HCC) however, its utility in canine HCC has not been explored. The aim of the study was to determine if PLR could predict survival outcomes in 42 dogs with HCC. PLR was not a significant predictive factor (p = 0.15) but lymphopenia alone was significantly correlated with a reduced probability of survival (p = 0.024). Further studies are needed to evaluate if peripheral lymphocyte count mirrors that of the tumor microenvironment in canine HCC.

Contribution of Hemogram Plan in the Horse's Clinical Evaluation.

The interpretation of the blood count is essential to help the equine clinician in the diagnosis, prognosis, patient management, and control of equine diseases. Hematologic alterations often reflect the condition of the individual or an overall response to a pathological situation. A thorough clinical examination of the patient is essential to correctly interpret the hematological results. The most common abnormalities in the erythrogram are mainly anemia and polycythemia. The frequent causes of anemia in horses are acute and chronic blood loss, hemolytic anemia, and anemia caused by chronic disease. Evaluation of leukogram, including a total white cell count, a differential cell count, absolute numbers of specific leukocytes can help identify abnormalities that may suggest specific diseases such as a viral or bacterial infection, inflammatory disorders or even a neoplastic process. The platelet count is most often used to monitor or diagnose conditions that cause too much bleeding related with thrombocytopenia; it can be due to multiple mechanisms such as reduction of thrombopoiesis (myeloptisis, myelofibrosis, myeloproliferative disease, and idiopathic medullary aplasias or due to the effect of mielosuppresive drugs), increased peripheral destruction of platelets (immune mediated thrombocytopenia), consumption (dissemined intravascular coagulation) sequestration of the spleen and loss of platelets by idiopathic origin.

Investigation of platelet measurands in dogs with hematologic neoplasia.

INTRODUCTION: Thrombocytopenia has been associated with some neoplastic processes, including hematologic neoplasia. There is no information regarding specific changes in platelet measurands in dogs with hematologic neoplasia compared with healthy dogs. The objectives of our study were to establish RIs, evaluate platelet measurands in dogs with hematologic neoplasia, and compare these measurands in patients with hematologic malignancies with or without thrombocytopenia. METHODS: This was a retrospective study. Platelet measurands were determined using the ADVIA 120 Hematology analyzer when a CBC was performed and included the platelet count, MPV, platelet distribution width (PDW), plateletcrit (PCT), mean platelet component (MPC), platelet component distribution width (PCDW), mean platelet mass (MPM), platelet mass distribution width (PMDW), and number of large platelets. Reference intervals were determined retrospectively using data from 129 healthy dogs. Patients with hematologic neoplasia (n = 50) were identified through retrospective evaluation of medical records from the Auburn University Veterinary Teaching Hospital and separated into thrombocytopenic (n = 20) and nonthrombocytopenic groups (n = 30). RESULTS: Platelet count and PCT were significantly higher in older healthy dogs compared with younger dogs. Significant differences were identified when comparing healthy dogs with those with hematologic neoplasia without thrombocytopenia for PDW, PCDW, PMDW, and the number of large platelets, indicating the presence of more heterogeneous platelets. Thrombocytopenic dogs with hematologic neoplasia had significantly decreased MPCs and increased MPVs, MPMs, and PCDWs compared with nonthrombocytopenic dogs with neoplasia. CONCLUSIONS: Dogs with hematologic neoplasia had more heterogeneous platelets, whereas thrombocytopenic patients with neoplasia had more activated platelets.

Amikacin prevents platelet aggregation in feline venous blood samples.

INTRODUCTION: Physiologically, feline platelets are more reactive and prone to aggregation, which interferes with platelet counts using automated counters and manual methods. The use of aminoglycoside amikacin in association with EDTA has proven to be efficient in preventing platelet aggregates in cases of pseudo thrombocytopenia (PTP) in people. OBJECTIVES: This study evaluated the efficacy of amikacin in preventing platelet aggregation in EDTA-containing feline blood samples and investigated the possible effects on hematologic measurands. MATERIALS AND METHODS: Blood samples (1.0 mL) collected from 100 healthy cats were stored in two EDTA tubes: 0.5 mL in a microtube containing 10 µL of 250 mg/mL amikacin (EDTA-AMK group) and 0.5 mL in a microtube containing only K2 EDTA 10% (EDTA group). A CBC was executed with an automated impedance blood analyzer, and a microscopic examination of the blood smears was performed. RESULTS: Platelet clumps were observed in 56% of samples from the EDTA group and 5% of samples from the EDTA-AMK group. Platelet counts (PLT), plateletcrit (PCT), and WBC counts were significantly higher (P < .001) in the EDTA-AMK group compared withi the EDTA group. CONCLUSIONS: Amikacin prevents platelet aggregation in feline venous blood samples and does not cause clinically relevant changes in other hematologic measurands. To our knowledge, this is the first report showing the use of amikacin in preventing platelet aggregation in feline blood samples. Based on this study, amikacin could be added to EDTA collection tubes for complete blood counts in cats.

Interleukin-6 and thrombopoietin concentrations in dogs with carcinoma with and without thrombocytosis.

BACKGROUND: Carcinoma-associated thrombocytosis involves tumor production of mediators such as interleukin-6 (IL-6) and thrombopoietin (TPO) that increase thrombopoiesis and may play a role in tumor evasion and metastasis. Carcinoma-associated thrombocytosis is described in people, but has not been described in dogs. HYPOTHESIS/OBJECTIVES: Evaluate the concentrations of IL-6 and TPO in dogs diagnosed with carcinoma with or without thrombocytosis. We hypothesized that IL-6 and TPO concentrations would be higher in dogs with carcinoma compared to healthy dogs, and that IL-6 and TPO concentrations would be higher in dogs with carcinoma and thrombocytosis when compared to dogs with carcinoma and normal platelet counts. ANIMALS: One-hundred sixteen dogs: 63 with carcinoma and 53 healthy control dogs. METHODS: Complete blood count was performed in all dogs, and they were stratified for sub-group analysis based on the presence or absence of thrombocytosis (platelet count > 500 103/µL). Serum TPO and IL-6 concentrations were measured by ELISA. Results of selected numeric variables were compared using Wilcoxon rank sum tests for pairwise comparisons. A value of P < .05 was considered significant. RESULTS: Twelve of the dogs with carcinoma (12/63, 19.0%) and none of the healthy control dogs (0%) had thrombocytosis. Thrombopoietin concentrations (median [range]) were significantly higher in dogs with carcinoma when compared to controls (87.42 pg/mL [0 to >600] vs 15.99 pg/mL [0 to >600], P < .001). Interleukin-6 concentrations (median [range]) were not different between dogs with carcinoma and healthy control dogs (9.70 pg/mL [0-181.53] vs 3.03 pg/mL [0-280.77], P = .15). In dogs with carcinoma, the TPO and IL-6 concentrations were not different between dogs with thrombocytosis and dogs with normal platelet count. CONCLUSIONS AND CLINICAL IMPORTANCE: Thrombopoietin concentrations were significantly higher in dogs with carcinoma, regardless of platelet count. Thrombopoietin is likely to be 1 of multiple factors that can impact platelet number, production, and consumption in dogs with carcinoma.

Neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios in dogs and cats with acute pancreatitis.

BACKGROUND: Neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been described as prognostic markers in cancer and inflammatory diseases. The use of these measurements in dogs and cats with acute pancreatitis has not yet been investigated. OBJECTIVE: The objective was to evaluate NLR and PLR for usefulness as biomarkers for severity and clinical course of acute pancreatitis in dogs and cats. METHODS: In this prospective study, we investigated 67 dogs and 41 cats with acute pancreatitis and 17 healthy dogs and cats as controls. We compared the NLR and PLR of dogs and cats with acute pancreatitis with control animals. The severity of disease was measured using a modified "bedside index of severity in acute pancreatitis" (BISAP) score in humans, and the course of the disease was measured with the time to clinical recovery. RESULTS: NLR and PLR were significantly higher in dogs and cats with pancreatitis than in healthy controls. However, no significant relationship between NLR or PLR and disease severity was found, as assessed with the modified BISAP score. Dogs and cats with pancreatitis and prolonged recovery displayed a significantly increased PLR, and those same cats also displayed a significantly increased NLR. CONCLUSIONS: We conclude that increased NLR and PLR provide information regarding the course of pancreatitis in dogs and cats.

Assessment of platelet biology in equine patients with systemic inflammatory response syndrome.

In addition to maintaining hemostasis, platelets have an important role in modulating innate and adaptive immune responses. A low platelet count has been found to be a negative prognostic factor for survival in humans and horses with critical illnesses, such as sepsis or systemic inflammatory response syndrome (SIRS). Decreased platelet aggregation, caused by in vivo activation, has been found in human patients with severe sepsis. In our prospective controlled study, we assessed platelet biology in blood samples from 20 equine SIRS cases and 120 healthy control horses. Platelet variables such as platelet count, large platelet count, clumps, plateletcrit, mean platelet volume, and mean platelet component concentration were analyzed by laser flow cytometry (Advia 2120) from K3EDTA blood and from citrate blood. Hirudin blood samples were analyzed by impedance aggregometry (Multiplate analyzer; Roche) for platelet aggregation, including spontaneous aggregation and aggregation by 4 different agonists: adenosine diphosphate (ADPtest), ADP + prostaglandin E1 (ADPtestHS), arachidonic acid (ASPItest), and collagen (COLtest). SIRS cases had significantly lower platelet counts in K3EDTA blood (p < 0.0001) compared to control horses. There were no significant differences in aggregation values between SIRS cases and controls. Non-surviving SIRS horses did not have statistically significant lower platelet counts or lower aggregation values for COLtest, ADPtest, or ADPtestHS compared to surviving SIRS horses, although 5 non-survivors were thrombocytopenic.

Evaluation of platelet indices and complete blood count in canine mammary tumors

Malignant mammary tumors in humans and bitches cause hematological disorders such as anemia, erythrocytosis, thrombocytosis, hyperproteinemia, and leucopenia. Novel studies have been conducted on the predictive and prognostic values of platelet (PLT) indices in human breast cancer (HBC). However, there is little information about the alterations in hematological parameters in canine mammary tumors (CMTs). The aims of this study were to evaluate the platelet indices and complete blood count (CBC) parameters in bitches with and without mammary tumor and to assess the above mentioned parameters with regard to histological tumor types and grades. A total of 71 bitches were enrolled in this study. The bitches in the study group were divided into 2 groups which consisted of malignant epithelial mammary tumors (group EMT; n = 43) and malignant mixed mammary tumors (group MMT; n = 12). Control group (group C) consisted of clinically and gynaecologically healthy 16 bitches. Blood samples were obtained to perform the CBC and PLT indices analysis. Histopathological examinations were carried out under a light microscope. Histological tumor types and malignancy grades were classified. The bitches with mammary tumor showed significantly increased PLT values and decreased hematocrit (HCT), hemoglobin (HGB) and mean corpuscular hemoglobin (MCH) values versus the healthy ones, regardless of the tumor type. However, in comparisons with the group C, mean platelet volume (MPV) and mean corpuscular hemoglobin concentration (MCHC) values were different only in the group MMT, while plateletcrit (PCT) and mean corpuscular volume (MCV) values were different only in the group EMT. Also white blood cell (WBC), PLT and PCT values were higher than the referenced laboratory ranges in grade 3 tumors. In the presented study, MPV was considerably correlated with PLT, platelet distribution width (PDW) and PCT. Also, PCT and PLT had high sensitivity and specificity to distinct EMT and MMT from the healthy bitches. Microcytic and hypochromic anemia occurs due to the decrease in the amount of HGB. Levels of MCV, MCH, and MCHC in the HBC group were reported to be significantly lower than in humans without breast cancer. Although anemia did not occur in EMT and MMT groups, obtained significances in the HCT, HGB, MCV, MCH, and MCHC levels between the bitches with and without mammary tumor were in line with the previous reports. In this study, WBC levels in grade 3 tumors were significantly higher than grade1 tumors (P < 0.05). Whereas levels of WBC in grade 1 and grade 2 tumors were in referenced laboratory ranges, it was higher in grade 3. Increased level of WBC in grade 3 was supposed to be due to the rise in malignancy as previously reported. Thrombocytosis was detected in 48.83% and 41.66% of the bitches in EMT and MMT groups, respectively. The higher percentage of CMTs with thrombocytosis in this study might be due to the difference in referenced upper limit of PLT in previous studies. The elapsed time between tumor formation and clinical presentation could be another influencing factor. Although PLT and PCT values were not significant according to the histological grading in this study, both parameters were found to be higher in grade 3 than the normal reference values. Further studies conducted with higher populations may lead the differences in these parameters to significance. With the support of further studies, alterations in the above mentioned parameters in bitches may contribute in the diagnosis process, management of treatment and may constitute an easy way to have an idea about the prognosis of mammary tumors.(AU)

Effect of siltuximab, omalizumab, infliximab, pembrolizumab and vedolizumab on selected haematological and biochemical parameters in a pig model.

From the regulatory point of view a strong link between an animal model and human pharmacodynamics of biological drugs is very important to qualify the model as "relevant". Consistent changes in cell population between human physiology and animal model gain value of this model which then can be pharmacodynamically "relevant" from the regulatory point of view. Consequently, the aim of this study was to determine how similar to human observations is the effect of selected biological drugs on blood cells in a pig model. The study was to carry out a comparative analysis of the variability of selected biochemical and hematological parameters of the blood after administration of five different human therapeutic monoclonal antibodies (mAbs) after a single subcutaneous (SC) dose in breeding pigs. The tested drugs were siltuximab (Syl- vant®), omalizumab (Xolair®), infliximab (Inflectra®), pembrolizumab (Keytruda®), and vedoli- zumab (Entyvio®) given in a single 1 mg/kg SC injection. Each of the tested drugs exerted a sig- nificant effect on at least two of the tested parameters three weeks after the administration. Siltuximab significantly influenced 9 of the analyzed parameters. Vedolizumab significantly influenced 8 of the analyzed parameters. Infliximab had the lowest impact of all the tested drugs, as it significantly influenced only two of the analyzed parameters. The study has proved that the impact of mAbs on the analyzed parameters can be significantly extended over time. This requires the monitoring of hematological parameters in the pig model even many weeks after administration of a drug in a relatively small dose.

Evaluation of the effect of vincristine on platelet count in dogs with lymphoma.

OBJECTIVES: To determine the effect of vincristine administration on the platelet count, platelet morphology and incidence of thrombocytopenia in dogs diagnosed with lymphoma. MATERIAL AND METHODS: Medical records of 59 dogs with lymphoma receiving vincristine sulphate were reviewed retrospectively. RESULTS: Following vincristine administration the platelet count was higher and the number of thrombocytopenic patients was lower. No difference was found in the number of dogs with enlarged and elliptical platelets following vincristine treatment. CLINICAL SIGNIFICANCE: Vincristine administration increases platelet counts in dogs with lymphoma. It is not contra-indicated to administer vincristine to dogs with lymphoma that are thrombocytopenic.

Immune thrombocytopenia (ITP): Pathophysiology update and diagnostic dilemmas.

Immune thrombocytopenia (ITP) is a common autoimmune bleeding disorder. The understanding of ITP pathogenesis is rapidly evolving. We now recognize ITP as a complex and heterogeneous syndrome that results from a combination of humoral and cell-mediated attacks on platelets peripherally and megakaryocytes in the bone marrow. Autoantibody-mediated ITP also varies in the pathway used to clear platelets, which depends on the platelet glycoprotein being targeted. Moreover, ITP patients present with variable bleeding severities and treatment responses that do not closely correlate with platelet count. A gold standard diagnostic test for ITP is lacking, and biomarkers to assess disease severity are in their infancy. This review provides an update on the immunopathogenesis of ITP and summarizes currently available tests for ITP diagnosis, prediction of disease severity, and treatment responses. Given the heterogeneous pathogenesis and clinical presentation of ITP, we highlight the need for the development of diagnostic and prognostic tests that would allow for the individualized management of a complex disease.

Urine 5-hydroxyindoleacetic acid in Cavalier King Charles spaniels with preclinical myxomatous mitral valve disease.

Higher concentrations of circulating serotonin have been reported in Cavalier King Charles spaniels (CKCS) compared to other dog breeds. The CKCS is also a breed highly predisposed to myxomatous mitral valve disease (MMVD). The aim of this study was to determine urine concentrations of 5-hydroxyindoleacetic acid (5-HIAA), the major metabolite and excretion product of serotonin, in a population of CKCS with preclinical MMVD, and to evaluate whether urine 5-HIAA concentrations were associated with MMVD severity, dog characteristics, setting for urine sampling, platelet count, and serotonin concentration in serum and platelet-poor plasma (PPP). The study population consisted of 40 privately-owned CKCS (23 females; 17 males) with and without preclinical MMVD as follows: American College of Veterinary Internal Medicine (ACVIM) group A (n = 11), ACVIM group B1 (n = 21) and ACVIM group B2 (n = 8). Urine 5-HIAA concentrations were not significantly associated with preclinical MMVD disease, platelet count or circulating concentrations of serotonin (in serum and PPP; P > 0.05). Females had higher 5-HIAA concentrations than males in morning urine collected at home (females, 3.1 [2.9-3.7] µmol/mmol creatinine [median and quartiles]; males, 1.7 [1.2-2.2] µmol/mmol creatinine; P = 0.0002) and urine collected at the clinic (females, 3.5 [3.1-3.9] µmol/mmol creatinine; males, 1.6 [1.3-2.1] µmol/mmol creatinine; P < 0.0001). Five-HIAA concentrations in urine collected at home and at the clinic were significantly associated (P = 0.0004; r = 0.73), and higher concentrations were found in urine collected at the clinic (P = 0.013). Urine 5-HIAA concentration was influenced by sex and setting of urine sampling. Urine 5-HIAA concentration was not associated with MMVD severity or circulating concentrations of serotonin in CKCS with preclinical disease.

Protocols for preparation of platelet rich plasma (PRP) in Quarter Horses / Protocolos para o preparo de plasma rico em plaquetas (PRP) em cavalos Quarto de Milha

This study compared two protocols for preparation of platelet rich plasma (PRP) and evaluated the association between manual and automated methods for platelet count using a prospective study design. Eight clinically healthy Quarter Horses had venous blood samples collected at rest. After collection, blood samples were centrifuged twice, using two different protocols including a period of sample resting, either at the start or at the end of the protocol. Platelet counting at the start of the protocol, during, and after obtaining PRP was conducted manually or with an automated counter, followed by comparison of the two methods. In order to investigate platelet degranulation during the protocol, vascular endothelial growth factor (VEGF) was measured at each preparation stage. The protocol with sample resting before centrifugation yielded a more concentrated PRP, and the study verified that both manual and automated methods are comparable and can be used interchangeably for platelet counting. VEGF concentration did not differ significantly between protocols, or among protocol stages. The results indicate that choice of protocol for PRP preparation will affect the quantity of platelets in the final product, although platelet degranulation was not observed as evidenced by the stable VEGF concentrations measured. A larger yield of non-degranulated platelets in PRP is desirable since more α-granules will be present, therefore Protocol II is recommended. Both manual and automated counts reliably allow clinicians to obtain platelet counts and the choice of utilizing a manual or automated method is unlikely to interfere with evaluation of the final PRP product.(AU)

Este estudo comparou dois protocolos de preparo de plasma rico em plaquetas (PRP) e avaliou a associação entre dois métodos de contagem plaquetária - um manual e o outro automático através de um estudo prospectivo. Sangue venoso de oito equinos da raça Quarto de Milha foi coletado e em seguida foi centrifugado duas vezes utilizando-se dois protocolos distintos: um com descanso antes da primeira centrifugação e outro após a segunda centrifugação. A contagem plaquetária ao início, no meio e ao final dos protocolos foi realizada manualmente e pelo método automatizado, seguida de comparação entre os dois métodos. Para investigar a degranulação plaquetária ocorrida durante o preparo do PRP, o fator de crescimento vascular endotelial (VEGF) foi mensurado em cada estágio dos protocolos. O método utilizando o descanso da amostra antes da primeira centrifugação proporcionou a obtenção de um PRP mais concentrado, além de o estudo verificar que ambos os métodos de contagem plaquetária (manual e automatizado) são comparáveis e podem ser usados indiferentemente. A concentração de VEGF não foi significativamente diferente entre os estágios de preparo do PRP. Os resultados indicam que o método de preparo afeta a quantidade de plaquetas obtidas no PRP, apesar da degranulação plaquetária não ter sido observada, como evidenciado pela concentração estável de VEGF. Uma maior concentração de plaquetas no PRP é desejável, pois indica que um maior número de α-grânulos estará presente na amostra, portanto, conclui-se que o Protocolo II é mais recomendável. Tanto o método manual, quanto o automatizado, pode ser usado de maneira confiável para a contagem plaquetária, não interferindo com a avaliação do produto final (PRP).(AU)

Predictive factors of malignancy in dogs with focal liver lesions using clinical data and ultrasonographic features.

A definitive diagnosis of focal liver lesions (FLLs) requires invasive procedures for histopathologic examination. Thus, a simpler noninvasive diagnostic method, such as conventional ultrasonography combined with clinical data, is needed for the prediction of liver malignancy. The objective of this study was to examine the diagnostic accuracy of clinical data and ultrasonographic (US) features to differentiate benign and malignant liver lesions. Medical records and US images from dogs with FLLs that underwent abdominal US and histopathologic examinations following surgery or liver biopsy were retrospectively reviewed. Clinical data, including signalment, clinical signs and laboratory findings, and the US features of liver lesions that could act as predictive factors were assessed using univariate and multivariate analyses to evaluate the associations between predictive factors and liver malignancy. Based on the histopathologic results, 55 dogs with malignant lesions and 28 dogs with benign lesions were included in the study. The results of univariate analysis showed that several US features and platelet count were significantly associated with liver malignancy. Multivariate analysis revealed that the platelet count (thrombocytosis; odds ratio [OR]: 4.13, 95% confidence intervals [CI]: 1.81-9.41), lesion size (4.1 cm or greater; OR: 23.83, 95% CI: 3.74-151.95) and echotexture of FLLs (heterogenous; OR: 8.44; 95% CI: 1.37-51.91) were independent predictors for differentiating benign and malignant liver lesions, suggesting that a combination of clinical data and US findings of FLLs could predict liver malignancy in dogs.

Comparison of Multiplate, Platelet Function Analyzer-200, and Plateletworks in Healthy Dogs Treated with Aspirin and Clopidogrel.

BACKGROUND: Platelet function testing may be warranted to assess response to aspirin and clopidogrel. HYPOTHESIS/OBJECTIVES: To evaluate the effects of aspirin, clopidogrel, or combination therapy using 3 platelet function tests: Multiplate Analyzer (MP), Platelet Function Analyzer-200 (PFA), and Plateletworks (PW). ANIMALS: Six healthy laboratory Beagles. METHODS: Randomized double-blind placebo-controlled study (crossover design). Dogs were given aspirin 1 mg/kg, clopidogrel 2 mg/kg, or combination therapy for 1 week each, with a washout period of 2 weeks. Platelet function was assessed on days 0 and 7 of each phase using MP (adenosine diphosphate [ADP], arachidonic acid [AA], collagen [COL] agonists), PFA (P2Y, COL-ADP [CADP], COL-Epinephrine [CEPI] cartridges), and PW (ADP, AA, COL agonists). Platelet counts were obtained with impedance and optical counters. RESULTS: For MP, mean aggregation was decreased for COL and AA with combination therapy and for ADP with all treatments. For PFA, mean CT was increased for the CEPI cartridge with aspirin; and for the P2Y and CADP cartridges with clopidogrel or combination therapy. More dogs receiving clopidogrel showed an increase in PFA CT using the P2Y than the CADP cartridge. For PW, mean aggregation was decreased for AA with all treatments; for ADP with clopidogrel or combination therapy; and for COL with clopidogrel. The PW results with the 2 hematology counters showed almost perfect agreement. CONCLUSION AND CLINICAL IMPORTANCE: All platelet function tests detected treatment effects in some dogs and may have utility for monitoring therapy.