Valores de Referência para tempos de coagulação em uma população de cães de Porto Alegre, RS, Brasil / Reference values for coagulation time of dogs in Porto Alegre, RS, Brazil

Background: Prothrombin time (PT) and activated partial thromboplastin time (PTT) are screening tests for coagulopathies in dogs. Both tests measure the clotting ability by the activation of different parts of the coagulation cascade. These tests vary widely in terms of reference parameters, mainly due the considerable diversity of reagents and analyzers available. In addition, there are many variations inherent to different populations, and little has been published about coagulation reference parameters for the local dog population. The main objective of the present study was to determine a clotting time reference range of a dog population in Porto Alegre, Rio Grande do Sul, Brazil.Materials, Methods & Results: Hemostatic reference range was determined from citrated plasma of 268 clinically heathy dogs of both genders. The animals did not present bleeding diathesis or thrombocytopenia history. All dogs were previously submitted to clinical examination (cardiopulmonary auscultation, abdominal palpation, and rectal temperature) and laboratory screening (complete blood count, creatinine, albumin, and alanine aminotransferase). PT and PTT of 71 and 258 samples were measured, respectively. Blood samples were collected into 2.7 mL 3.2 % sodium citrate tubes (9 parts blood : 1 part citrate) by vacuum. Blood samples were centrifuged; the plasma was harvested and stored at -30 °C upon analyses. All analyses were performed using the viscosity detection method in semi-automatic coagulometer according to manufacturer’s guidelines. The reference ranges were determined in accordance with the American Society for Veterinary Clinical Pathology. The PT and PTT reference ranges were between 6.06 to 9.32 s and between 15.25 to 24.57 s, respectively.Discussion: The increased activity of the extrinsic pathway clotting factor generates narrow values in the PT assay, in comparison to PTT results.[...]

Valores de Referência para tempos de coagulação em uma população de cães de Porto Alegre, RS, Brasil / Reference values for coagulation time of dogs in Porto Alegre, RS, Brazil

Background: Prothrombin time (PT) and activated partial thromboplastin time (PTT) are screening tests for coagulopathies in dogs. Both tests measure the clotting ability by the activation of different parts of the coagulation cascade. These tests vary widely in terms of reference parameters, mainly due the considerable diversity of reagents and analyzers available. In addition, there are many variations inherent to different populations, and little has been published about coagulation reference parameters for the local dog population. The main objective of the present study was to determine a clotting time reference range of a dog population in Porto Alegre, Rio Grande do Sul, Brazil.Materials, Methods & Results: Hemostatic reference range was determined from citrated plasma of 268 clinically heathy dogs of both genders. The animals did not present bleeding diathesis or thrombocytopenia history. All dogs were previously submitted to clinical examination (cardiopulmonary auscultation, abdominal palpation, and rectal temperature) and laboratory screening (complete blood count, creatinine, albumin, and alanine aminotransferase). PT and PTT of 71 and 258 samples were measured, respectively. Blood samples were collected into 2.7 mL 3.2 % sodium citrate tubes (9 parts blood : 1 part citrate) by vacuum. Blood samples were centrifuged; the plasma was harvested and stored at -30 °C upon analyses. All analyses were performed using the viscosity detection method in semi-automatic coagulometer according to manufacturers guidelines. The reference ranges were determined in accordance with the American Society for Veterinary Clinical Pathology. The PT and PTT reference ranges were between 6.06 to 9.32 s and between 15.25 to 24.57 s, respectively.Discussion: The increased activity of the extrinsic pathway clotting factor generates narrow values in the PT assay, in comparison to PTT results.[...](AU)

Avaliação do pool de plasma caseiro como controle normal para o tempo de tromboplastina parcial ativada (TTP) / Evaluation if in-house preparations of polled plasma can be used as a normal control of partial thromboplastin time (PTT)

INTRODUÇÃO: A utilização correta de um plasma-controle para a comparação com o plasma do paciente nos testes de coagulação é fundamental para a garantia de um resultado seguro dessas provas laboratoriais. OBJETIVO: O presente estudo analisou a viabilidade do uso de um pool de plasma caseiro, realizado com cinco (P5) e 20 (P20) amostras a partir de pacientes normais, para ser utilizado como controle normal do tempo de tromboplastina parcial (TTP). MATERIAL E MÉTODO: Os dois pools de plasma caseiro foram analisados em relação a um controle normal comercial (AP). Foram 10 dias de experimento e a cada dia os dois pools caseiro eram feitos. Para cada dia foi feito o TTP de P5, P20 e AP. Todos os pacientes com solicitação de TTP, em cada dia do experimento, tiveram a relação de tempos (R) determinada frente a P5, P20 e AP. As ferramentas estatísticas utilizadas foram média (X), análise de variância e teste de Tukey. RESULTADOS: A análise estatística demonstrou que os valores de TTP são significativamente diferentes entre AP e P5 e entre AP e P20, mas não há diferença significativa entre P5 e P20. Quando a relação de tempos foi analisada, não houve diferença significativa entre AP, P5 e P20. CONCLUSÃO: O estudo demonstrou que pode ser utilizado como controle normal um pool de plasma caseiro, feito a partir de cinco ou 20 amostras.

INTRODUCTION: Proper utilization of plasma control for comparison with the plasma of patients with coagulation tests is critical to ensure a safe outcome of these laboratory tests. OBJECTIVE: The purpose of this work was to know if in-house preparations of polled plasma can be used as a normal control of partial thromboplastin time (PTT). MATERIAL AND METHOD: We make two polled plasma, one with five (P5) samples and another with 20 (P20) samples, both of them from normal patients. Both pooled plasma were analyzed in comparison with a commercial lyophilized control (AP). The experiment was performed in 10 days, and P5 and P20 were made daily. At each day PTT was performed for P5, P20 and AP. All patients who asked for TTP, in each day of this experience, got a time relation (R) determined by a P5, P20 e AP. Tukey test was used as statistical analysis. RESULTS: Statistically significant differences were detected among the PTT for AP and P5, AP and P20, but no difference between P5 and P20 was observed. When the time relations were tested, there were no significant difference, among AP, P5, and P20. CONCLUSION: We found that in-house preparations of polled plasma (P5 and P20) can be used as a normal control of PTT.

Biological potency evaluation and physicochemical characterization of unfractionated heparins

Unfractionated heparins are used clinically as anticoagulants. The biological potency of thirteen samples of raw material and pharmaceutical formulations were assessed utilizing the 5th International Standard of heparin using the sheep plasma coagulation inhibition assay, activated partial thromboplastin time, anti-factor Xa assay, and anti-factor IIa assay, resulting in mean potencies of 101.15 percent, 96.15 percent, 98.15 percent and 99.37 percent, respectively. The samples were also evaluated by the protamine neutralization test giving results within the range of 92 - 138 IU/mg. The anti-factor IIa assay was performed showing reproducibility and significant correlation with the pharmacopoeial assays, thus demonstrating it to be a feasible alternative to the sheep plasma coagulation inhibition assay. Moreover, an analysis by nuclear magnetic resonance and capillary electrophoresis showed some peaks attributable to oversulfated chondroitin sulfate. The results show that batch-to-batch variations and the quality of samples contributed to improvements in the quality control of pharmaceutical products and to assure the safe use and clinical efficacy of this biological medicine.

As heparinas não fracionadas são utilizadas clinicamente como anticoagulantes. A potência biológica de 13 amostras de matérias-primas e produtos farmacêuticos foram avaliadas em relação ao 5ª Padrão Internacional de heparina pelos ensaios da inibição da coagulação do plasma ovino, tempo de tromboplastina parcial ativada, anti-fator Xa e anti-fator IIa, que forneceram potências médias de 101,15 por cento, 96,15 por cento, 98,15 por cento e 99,37 por cento, respectivamente. As amostras foram também submetidas ao teste de neutralização pela protamina que apresentou resultados entre 92-138 UI/mg. Demonstrou-se reprodutibilidade e correlação significativa do ensaio do anti-fator IIa com os farmacopeicos, constituindo-se em alternativa ao ensaio da inibição da coagulação do plasma ovino. Além disso, as análises realizadas por ressonância magnética nuclear e eletroforese capilar mostraram picos correspondentesà condroitina supersulfatada. Os resultados mostraram variações lote-a-lote e a qualidade das amostras contribuindo para aprimorar o controle de qualidade dos produtos farmacêuticos e garantir a segurança e eficácia terapêutica desses produtos biológicos.